From 21496243e50c253a8f8e7207e888d373c2cbfc0e Mon Sep 17 00:00:00 2001 From: Pengyi Date: Thu, 19 Mar 2015 10:54:43 -0400 Subject: [PATCH 01/12] Update README.md --- README.md | 11 +++++++++-- 1 file changed, 9 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index 91273e2..c77c7a5 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,11 @@ -directPA +### Direction analysis for pathways and kinases ======== -A package for directional pathway analysis in experiments with multiple perturbation designs. +#### Description +`directPA' is a package designed for analyzing (1) gene/protein expression data using microarray, RNA-seq, or proteomics data, and (2) kinase enrichment using phosphoproteomics data. It intergrates multiple experimental treatments and test the effect of each combination of treatments using a directional approach. + +#### Download and install +The relsease version can be downloaded from CRAN [link](http://cran.r-project.org/web/packages/directPA/); + +Install the release version from CRAN with `install.packages("directPA")` + From f1465e870f3850a1a44d3e5de4d9cd64441bbf19 Mon Sep 17 00:00:00 2001 From: Pengyi Date: Thu, 19 Mar 2015 10:59:15 -0400 Subject: [PATCH 02/12] Update README.md --- README.md | 25 +++++++++++++++++++++++++ 1 file changed, 25 insertions(+) diff --git a/README.md b/README.md index c77c7a5..551fb49 100644 --- a/README.md +++ b/README.md @@ -9,3 +9,28 @@ The relsease version can be downloaded from CRAN [link](http://cran.r-project.or Install the release version from CRAN with `install.packages("directPA")` +#### Examples +I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +```r +# load the phosphoproteomics dataset +data(HEK) + +# load the kinase-substrate annoations +data(PhosphoSite) + +# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +# (180 degree to original direction) +kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +kst1[order(unlist(kst1[,1])),][1:20,] + +# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +# (135 degree to the original direction) +kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +kst2[order(unlist(kst2[,1])),][1:20,] + +# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +# (225 degree to the original direction) +kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +kst3[order(unlist(kst3[,1])),][1:20,] +``` From 146247a835ec98adb76da6fa90435c69cfe2c0bf Mon Sep 17 00:00:00 2001 From: PengyiYang Date: Tue, 17 Nov 2015 15:52:08 +1100 Subject: [PATCH 03/12] updates 17 Nov 2015 --- .gitignore | 3 + source/directPA_1.2/.Rbuildignore | 2 + source/directPA_1.2/.gitignore | 3 + source/directPA_1.2/DESCRIPTION | 14 ++ source/directPA_1.2/MD5 | 30 ++++ source/directPA_1.2/NAMESPACE | 11 ++ source/directPA_1.2/R/directExplorer2d.R | 93 ++++++++++++ source/directPA_1.2/R/directPA.R | 139 ++++++++++++++++++ source/directPA_1.2/R/geneStats.R | 43 ++++++ source/directPA_1.2/R/pathwayStats.R | 63 ++++++++ source/directPA_1.2/R/perturbPlot2d.R | 67 +++++++++ source/directPA_1.2/R/rotate2d.R | 26 ++++ source/directPA_1.2/R/rotate3d.R | 44 ++++++ source/directPA_1.2/data/HEK.rda | Bin 0 -> 31259 bytes source/directPA_1.2/data/PM.rda | Bin 0 -> 27357 bytes source/directPA_1.2/data/Pathways.rda | Bin 0 -> 86220 bytes source/directPA_1.2/data/PhosphoELM.rda | Bin 0 -> 18627 bytes source/directPA_1.2/data/PhosphoSite.rda | Bin 0 -> 72692 bytes source/directPA_1.2/directPA.Rproj | 17 +++ source/directPA_1.2/man/HEK.Rd | 8 + source/directPA_1.2/man/PM.Rd | 8 + source/directPA_1.2/man/Pathways.KEGG.Rd | 7 + source/directPA_1.2/man/Pathways.reactome.Rd | 7 + source/directPA_1.2/man/PhosphoELM.human.Rd | 9 ++ source/directPA_1.2/man/PhosphoELM.mouse.Rd | 9 ++ source/directPA_1.2/man/PhosphoSite.human.Rd | 8 + source/directPA_1.2/man/PhosphoSite.mouse.Rd | 8 + source/directPA_1.2/man/directEnrichPlot2d.Rd | 38 +++++ source/directPA_1.2/man/directExplorer2d.Rd | 60 ++++++++ source/directPA_1.2/man/directPA-package.Rd | 29 ++++ source/directPA_1.2/man/directPA.Rd | 93 ++++++++++++ source/directPA_1.2/man/geneStats.Rd | 36 +++++ source/directPA_1.2/man/pathwayStats.Rd | 50 +++++++ source/directPA_1.2/man/rotate2d.Rd | 31 ++++ source/directPA_1.2/man/rotate3d.Rd | 31 ++++ 35 files changed, 987 insertions(+) create mode 100644 .gitignore create mode 100644 source/directPA_1.2/.Rbuildignore create mode 100644 source/directPA_1.2/.gitignore create mode 100644 source/directPA_1.2/DESCRIPTION create mode 100644 source/directPA_1.2/MD5 create mode 100644 source/directPA_1.2/NAMESPACE create mode 100644 source/directPA_1.2/R/directExplorer2d.R create mode 100644 source/directPA_1.2/R/directPA.R create mode 100644 source/directPA_1.2/R/geneStats.R create mode 100644 source/directPA_1.2/R/pathwayStats.R create mode 100644 source/directPA_1.2/R/perturbPlot2d.R create mode 100644 source/directPA_1.2/R/rotate2d.R create mode 100644 source/directPA_1.2/R/rotate3d.R create mode 100644 source/directPA_1.2/data/HEK.rda create mode 100644 source/directPA_1.2/data/PM.rda create mode 100644 source/directPA_1.2/data/Pathways.rda create mode 100644 source/directPA_1.2/data/PhosphoELM.rda create mode 100644 source/directPA_1.2/data/PhosphoSite.rda create mode 100644 source/directPA_1.2/directPA.Rproj create mode 100644 source/directPA_1.2/man/HEK.Rd create mode 100644 source/directPA_1.2/man/PM.Rd create mode 100644 source/directPA_1.2/man/Pathways.KEGG.Rd create mode 100644 source/directPA_1.2/man/Pathways.reactome.Rd create mode 100644 source/directPA_1.2/man/PhosphoELM.human.Rd create mode 100644 source/directPA_1.2/man/PhosphoELM.mouse.Rd create mode 100644 source/directPA_1.2/man/PhosphoSite.human.Rd create mode 100644 source/directPA_1.2/man/PhosphoSite.mouse.Rd create mode 100644 source/directPA_1.2/man/directEnrichPlot2d.Rd create mode 100644 source/directPA_1.2/man/directExplorer2d.Rd create mode 100644 source/directPA_1.2/man/directPA-package.Rd create mode 100644 source/directPA_1.2/man/directPA.Rd create mode 100644 source/directPA_1.2/man/geneStats.Rd create mode 100644 source/directPA_1.2/man/pathwayStats.Rd create mode 100644 source/directPA_1.2/man/rotate2d.Rd create mode 100644 source/directPA_1.2/man/rotate3d.Rd diff --git a/.gitignore b/.gitignore new file mode 100644 index 0000000..807ea25 --- /dev/null +++ b/.gitignore @@ -0,0 +1,3 @@ +.Rproj.user +.Rhistory +.RData diff --git a/source/directPA_1.2/.Rbuildignore b/source/directPA_1.2/.Rbuildignore new file mode 100644 index 0000000..91114bf --- /dev/null +++ b/source/directPA_1.2/.Rbuildignore @@ -0,0 +1,2 @@ +^.*\.Rproj$ +^\.Rproj\.user$ diff --git a/source/directPA_1.2/.gitignore b/source/directPA_1.2/.gitignore new file mode 100644 index 0000000..807ea25 --- /dev/null +++ b/source/directPA_1.2/.gitignore @@ -0,0 +1,3 @@ +.Rproj.user +.Rhistory +.RData diff --git a/source/directPA_1.2/DESCRIPTION b/source/directPA_1.2/DESCRIPTION new file mode 100644 index 0000000..c66409c --- /dev/null +++ b/source/directPA_1.2/DESCRIPTION @@ -0,0 +1,14 @@ +Package: directPA +Type: Package +Title: Direction Analysis for Pathways and Kinases +Version: 1.2 +Date: 2015-06-18 +Author: Pengyi Yang & Ellis Patrick +Maintainer: Pengyi Yang +Description: Direction analysis is a set of tools designed to integrate multiple experimental treatments for analyzing pathways and kinases from microarray, RNA-seq, proteomics, or phosphoproteomics data. +License: GPL-3 +Depends: R (>= 2.10.0), rgl, calibrate +Packaged: 2015-06-18 13:08:32 UTC; Pengyi +NeedsCompilation: no +Repository: CRAN +Date/Publication: 2015-06-18 18:00:22 diff --git a/source/directPA_1.2/MD5 b/source/directPA_1.2/MD5 new file mode 100644 index 0000000..3c8c935 --- /dev/null +++ b/source/directPA_1.2/MD5 @@ -0,0 +1,30 @@ +e05721202d78707adb25b5dadb9a905c *DESCRIPTION +32f7448fafd8f272e98f8f3053cc3f51 *NAMESPACE +61911534dedf36247467a549fbf57ec1 *R/directEnrichPlot2d.R +76cad031899e402b3ab2c8ee3bad588f *R/directExplorer2d.R +c193001c14dbc241fd64be4f11202254 *R/directPA.R +016bae5eb003dc005c4dd37ccc50f58e *R/geneStats.R +c94800bbd93527de53b09fc6bbce19a3 *R/pathwayStats.R +0fb8763a7e2602a1004522f451bbea85 *R/rotate2d.R +989f12754c30a855e238dd4a160a9806 *R/rotate3d.R +83648a7a500619337f5f19913ad14f71 *data/HEK.rda +079cc541e6cfd73ded8ab5b20a8687e6 *data/PM.rda +d8b015f9de0c68ca738fa61b8604add3 *data/Pathways.rda +a4829f119c5d601a1663ff68b6f033c9 *data/PhosphoELM.rda +48bf3aaf03017c2519d62984ea9587a2 *data/PhosphoSite.rda +a8abd01dded8fe74481dfc5d6ee6284a *man/HEK.Rd +c02262b58e37b1fefd695441707b8809 *man/PM.Rd +19d4e0276d10cd41f6d577ed5245ea12 *man/Pathways.KEGG.Rd +ffd89f58f9af47d0ff556011a5c3521c *man/Pathways.reactome.Rd +50f5b7321b62385a4af7e5c1ee1ee901 *man/PhosphoELM.human.Rd +8e9a9b800cbdb9e5e9764143c54ef6fb *man/PhosphoELM.mouse.Rd +9224f661163899dbf13b3b3a8775a167 *man/PhosphoSite.human.Rd +02df716ef54fb50fdecd8648c6883235 *man/PhosphoSite.mouse.Rd +3d1ba4de7361b2947991bf8b6f1de90b *man/directEnrichPlot2d.Rd +29b9f292ce686fa8368ab99a2a527a74 *man/directExplorer2d.Rd +8809fdbeedb03dd0c8672be14ef11383 *man/directPA-package.Rd +02069d7456d6905cc361baab683de7f8 *man/directPA.Rd +fcc77f5e75cdbce83340ec66f2777f47 *man/geneStats.Rd +aefc1a7e9de81ead1d8f8d1911002812 *man/pathwayStats.Rd +3e13753f1c19b7fb239f048db2376b20 *man/rotate2d.Rd +574ba8d8e806479f12b078b52b6de344 *man/rotate3d.Rd diff --git a/source/directPA_1.2/NAMESPACE b/source/directPA_1.2/NAMESPACE new file mode 100644 index 0000000..4cdaa62 --- /dev/null +++ b/source/directPA_1.2/NAMESPACE @@ -0,0 +1,11 @@ +# Generated by roxygen2 (4.1.0): do not edit by hand + +export(perturbPlot2d) +export(directExplorer2d) +export(directPA) +export(geneStats) +export(pathwayStats) +export(rotate2d) +export(rotate3d) +import(rgl) +import(calibrate) diff --git a/source/directPA_1.2/R/directExplorer2d.R b/source/directPA_1.2/R/directExplorer2d.R new file mode 100644 index 0000000..0d8e08b --- /dev/null +++ b/source/directPA_1.2/R/directExplorer2d.R @@ -0,0 +1,93 @@ +#' Batch Direction Analysis in 2-dimentional space +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", +#' path.method="Stouffer", top=10, ...) +#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns +#' are treatments vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param top the number of entries to be highlighted in the plot. +#' @param ... parameters for controlling the plot. +#' @return The the list of enrichment analysis in tables. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # test enrichment on 8 directions in polar coordinate system. +#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) +#' +#' # the direction are denoted as follow for the two treatments vs control: +#' # ++: up-regulated in both treatments +#' # +*: up-regulated in the first treatment and unchanged in the second treatment +#' # +-: up-regulated in the first treatment and down-regulated in the second treatment +#' # *-: unchanged in the first treatment and down-regulated in the second treatment +#' # --: down-regulated in both treatments +#' # -*: down-regulated in the first treatment and unchanged in the second treatment +#' # -+: down-regulated in the first treatment and up-regulated in the second treatment +#' # *+: unchanged in the first treatment and up-regulated in the second treatment +#' +#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both +#' # treatments (i.e. "--") and displa the top-10 entries +#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] +#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] +#' +directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { + + directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') + gene.tab <- matrix(NA, nrow(Tc), 8) + colnames(gene.tab) <- directionCode + rownames(gene.tab) <- rownames(Tc) + + path.tab <- NULL + if (!is.null(annotation)) { + path.tab <- matrix(NA, length(annotation), 9) + colnames(path.tab) <- c("size", directionCode) + rownames(path.tab) <- names(annotation) + } + + for(i in 0:7){ + # gene or phosphorylation site level + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + gene.tab[,(i+1)] <- gene.pvalues + + # pathway or kinase level + if (!is.null(annotation)) { + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + if (i == 0) { + path.tab[,1] <- unlist(gst[,"size"]) + } + path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) + } + } + + # highlight candidates + plot(Tc, col="gray", pch=16, ...) + abline(h=0, v=0, col="gold", lty=2) + abline(a=0, b=1, lty = 2, col="darkgreen") + color <- rainbow(8) + for (i in 1:8) { + ids <- names(sort(gene.tab[,i])[1:top]) + points(Tc[ids,], col=color[i], pch=16) + textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) + } + + results <- list() + results$gene.tab <- gene.tab + results$path.tab <- path.tab + return(results) +} + diff --git a/source/directPA_1.2/R/directPA.R b/source/directPA_1.2/R/directPA.R new file mode 100644 index 0000000..0e2b4e4 --- /dev/null +++ b/source/directPA_1.2/R/directPA.R @@ -0,0 +1,139 @@ +#' Direction Analysis for Pathways and Kinases +#' +#' The main function of direction Analysis. This function takes in a matrix of test statistics with +#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for +#' enrichment of pathways or kinases on the specified direction. +#' +#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +#' path.method="Stouffer", visualize=TRUE) +#' +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are +#' treatments vs control statistics. +#' @param direction the direction to be tested for enrichment. Either specified as a degree for +#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the +#' @return a list of enrichment for pathways or kinases +#' @export +#' @examples +#' +#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +#' # (180 degree to original direction) +#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +#' kst1[order(unlist(kst1[,1])),][1:20,] +#' +#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +#' # (135 degree to the original direction) +#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +#' kst2[order(unlist(kst2[,1])),][1:20,] +#' +#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +#' # (225 degree to the original direction) +#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +#' kst3[order(unlist(kst3[,1])),][1:20,] +#' +#' +#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +#' # load the proteomics dataset +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # display reactome pathways. Could be replaced by any other pathway databases +#' Pathways.reactome[1:5] +#' +#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +#' # on the original direction. +#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +#' gst1[order(unlist(gst1[,1])),][1:20,] +#' +#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +#' gst2[order(unlist(gst2[,1])),][1:20,] +#' +#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +#' gst3[order(unlist(gst3[,1])),][1:20,] +#' +directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE, ...){ + ## spherical coordinates for three-dimensional rotation + if (length(direction) == 3) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate3d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot3d(Tc, col=HC[rank(gene.pvalues)], size=5, ...) + abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) + abclines3d(x=0, a=direction, col="pink", lwd=5) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } + + ## polar coordinates for two-dimensional rotation + if (length(direction) == 1) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate2d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot(Tc, col=HC[rank(gene.pvalues)], pch=16, ...) + abline(v = 0,h = 0, lty=2, col="gold") + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } +} diff --git a/source/directPA_1.2/R/geneStats.R b/source/directPA_1.2/R/geneStats.R new file mode 100644 index 0000000..2d296dc --- /dev/null +++ b/source/directPA_1.2/R/geneStats.R @@ -0,0 +1,43 @@ +#' Gene Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +#' and calculates a combined statistics accross multiple treatments. +#' +#' @usage geneStats(T, method="OSP") +#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +#' comparison. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. +#' @return a p-value after integration across treatments. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +geneStats <- function(T, method="OSP") { + pvalue <- 0 + + if (method == "Stouffer") { + pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) + } else if (method == "OSP") { + p <- pnorm(T, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(T, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(T), lower.tail = FALSE) + } + + return (pvalue) +} diff --git a/source/directPA_1.2/R/pathwayStats.R b/source/directPA_1.2/R/pathwayStats.R new file mode 100644 index 0000000..107581f --- /dev/null +++ b/source/directPA_1.2/R/pathwayStats.R @@ -0,0 +1,63 @@ +#' Pathway Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +#' and calculates a combined statistics for those that belong to the same pathway or kinase. +#' +#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") +#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. +#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or +#' phosphorylation site that belong to the same pathway or kinase. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. +#' @return a doublet corresponding to the enrichment after integration across all genes or substrates +#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +#' or kinase. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +#' # compute statistics for all reactome pathways +#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +#' +pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ + pvalue <- 0 + Z <- T[names(T) %in% PGs] + + if (length(Z) >= minSize) { + if (method == "Stouffer") { + pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) + } else if (method == "OSP") { + p <- pnorm(Z, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(Z, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(Z), lower.tail = FALSE) + } + } else { + pvalue <- NA + } + + result <- list() + result$pvalue <- pvalue + result$size <- length(Z) + return (result) +} diff --git a/source/directPA_1.2/R/perturbPlot2d.R b/source/directPA_1.2/R/perturbPlot2d.R new file mode 100644 index 0000000..b46dcfb --- /dev/null +++ b/source/directPA_1.2/R/perturbPlot2d.R @@ -0,0 +1,67 @@ +#' Perturbation Plot +#' +#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the +#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment +#' of pathways or kinases in direction specific manner. +#' +#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments +#' vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or +#' substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param ... parameters for controling the plot. +#' @return a list of coordinates for pathways or kinases +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' perturbPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3) +#' +perturbPlot2d <- function(Tc, annotation, minSize=5, ...) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. filter the groups that are smaller than the minimun cutoff + DE = lapply(annotation, function(x){ + if(sum(rownames(Tc.zscores) %in% x) >= minSize) { + X <- Tc.zscores[rownames(Tc.zscores)%in%x,] + n = nrow(X) + Z1 = sum(X[,1])/sqrt(n) + Z2 = sum(X[,2])/sqrt(n) + list(Z1=Z1, Z2=Z2) + } + }) + + # step3. filter DE that has 0 element + DE <- DE[which(sapply(DE, length) != 0)] + Z1 <- unlist(sapply(DE, function(x){x[1]})) + Z2 <- unlist(sapply(DE, function(x){x[2]})) + + # visualization + plot(Z1, Z2, col="darkblue", pch=16, xlab=colnames(Tc)[1], ylab=colnames(Tc)[2], ...) + textxy(Z1,Z2, names(DE), col="black", cex=1) + abline(v=0, h=0, col="gold", lty=2) + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + + r <- ceiling(max(sqrt(Z1^2 + Z2^2))) + for(i in seq(0, r, r/5)) { + theta = seq(-3.14,3.14,0.05) + lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") + } + + points(Z1, Z2, col="darkblue", pch=16, cex=2) + + ## return the results + result <- list() + result$Z1 <- Z1 + result$Z2 <- Z2 + return(result) +} diff --git a/source/directPA_1.2/R/rotate2d.R b/source/directPA_1.2/R/rotate2d.R new file mode 100644 index 0000000..c6012ca --- /dev/null +++ b/source/directPA_1.2/R/rotate2d.R @@ -0,0 +1,26 @@ +#' Polar Coordinates Rotation +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage rotate2d(T, degree = 0) +#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # convert statistics into z-scores +#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) +#' +#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +#' +rotate2d = function(T, degree = 0){ + theta = atan2(T[,2], T[,1]) + degree + r = sqrt(T[,1]^2 + T[,2]^2) + x2 = r*cos(theta) + y2 = r*sin(theta) + return(cbind(x2, y2)) +} diff --git a/source/directPA_1.2/R/rotate3d.R b/source/directPA_1.2/R/rotate3d.R new file mode 100644 index 0000000..10f8a79 --- /dev/null +++ b/source/directPA_1.2/R/rotate3d.R @@ -0,0 +1,44 @@ +#' Spherical Coordinates Rotation +#' +#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +#' @usage rotate3d(T, contrast) +#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +rotate3d = function(T, contrast = c(1,1,1)){ + + a = contrast + b= c(1,1,1) # the original direction in contrast form is c(1,1,1) + + if(sum(a == b)==3)return(T) + if(sum(a == -b)==3)return(-T) + + a = a/sqrt(sum(a^2)) + b = b/sqrt(sum(b^2)) + u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) + u = u/sqrt(sum(u^2)) + R = matrix(0,3,3) + the = acos(sum(a*b)) + R[1,1] = cos(the) + u[1]^2*(1-cos(the)) + R[2,2] = cos(the) + u[2]^2*(1-cos(the)) + R[3,3] = cos(the) + u[3]^2*(1-cos(the)) + 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zL80D?idv`n{h{jM%DBslXiOX7kg=a*-#ZID{S`)(-bM}+M0!(Ou~M2 zR~=WKI4MDJQ9XLeQ60yM*7&TsFcBuz&nA+#MBfD*|H&^y zr>Ftr|HnqFz%IZd7p-kJVq7m6hs{K8Dg+=^R>izt$LLxn1T zh@^;~SvrT$N^04HCUD=*W0t=v+z}}usOBWS^IJqi63kKZXE=Szv5 zXXF`M+VnX|D-u%7=J8d=op487T9sFu8=%!{h^LnIrbLM)w?ominYr7KsC_vbo=fN4 zHi#fl&v)Mpq5GB;wNZ;hjh8u2_4+Vn%~DMiipZ&b4{2N%s~nO&hTd}%ag5;XMT3cv zOyx)+Po4~B=4p>#uOF-CjObeL-gNlG0b6Y?x4D7Q;NJVATq7yp#Olv|Cp$32tbR5M*AOU>Iap!gYL)N_6Oce$8}DY>jE#0$egd8h()CXp7<#x7 zJ;MLWMCb3nl+QXu^j>yt@l4wlD-KN*`u zzNBXx(L7_lt87iX zTsA0PW-^WwN4R1ixCJC7cY_}*%;igRvvLh|K0&q-xO2&}3JAazLJ_OIWH~>ZRdI0M z!KVIs`>{Zs2moe1^&wVwt?jx)UYg;;;NNX!tIPB0d?wZ``LQDC;{dP!cBtGvx}Bce zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, +David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} diff --git a/source/directPA_1.2/man/directPA.Rd b/source/directPA_1.2/man/directPA.Rd new file mode 100644 index 0000000..50ffbeb --- /dev/null +++ b/source/directPA_1.2/man/directPA.Rd @@ -0,0 +1,93 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/directPA.R +\name{directPA} +\alias{directPA} +\title{Direction Analysis for Pathways and Kinases} +\usage{ +directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +path.method="Stouffer", visualize=TRUE) +} +\arguments{ +\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are +treatments vs control statistics.} + +\item{direction}{the direction to be tested for enrichment. Either specified as a degree for +two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} + +\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to +genes or substrates belong to each pathway or kinase, respectively.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{gene.method}{the method to be used for integrating statistics across treatments for each gene +or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is OSP.} + +\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation +sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is Stouffer.} + +\item{visualize}{whether to visualize the} +} +\value{ +a list of enrichment for pathways or kinases +} +\description{ +The main function of direction Analysis. This function takes in a matrix of test statistics with +two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +the annotation list such as pathway annotation or kinase-substrate annotation, and test for +enrichment of pathways or kinases on the specified direction. +} +\examples{ +## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +# load the phosphoproteomics dataset +data(HEK) + +# load the kinase-substrate annoations +data(PhosphoSite) + +# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +# (180 degree to original direction) +kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +kst1[order(unlist(kst1[,1])),][1:20,] + +# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +# (135 degree to the original direction) +kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +kst2[order(unlist(kst2[,1])),][1:20,] + +# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +# (225 degree to the original direction) +kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +kst3[order(unlist(kst3[,1])),][1:20,] + + +## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +# load the proteomics dataset +data(PM) + +# load pathway annotations +data(Pathways) + +# display reactome pathways. Could be replaced by any other pathway databases +Pathways.reactome[1:5] + +# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +# on the original direction. +gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +gst1[order(unlist(gst1[,1])),][1:20,] + +# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +gst2[order(unlist(gst2[,1])),][1:20,] + +# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +gst3[order(unlist(gst3[,1])),][1:20,] +} + diff --git a/source/directPA_1.2/man/geneStats.Rd b/source/directPA_1.2/man/geneStats.Rd new file mode 100644 index 0000000..455e3ef --- /dev/null +++ b/source/directPA_1.2/man/geneStats.Rd @@ -0,0 +1,36 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/geneStats.R +\name{geneStats} +\alias{geneStats} +\title{Gene Level Statistics} +\usage{ +geneStats(T, method="OSP") +} +\arguments{ +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +comparison.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} +} +\value{ +a p-value after integration across treatments. +} +\description{ +Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +and calculates a combined statistics accross multiple treatments. +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) +} + diff --git a/source/directPA_1.2/man/pathwayStats.Rd b/source/directPA_1.2/man/pathwayStats.Rd new file mode 100644 index 0000000..fec4ea5 --- /dev/null +++ b/source/directPA_1.2/man/pathwayStats.Rd @@ -0,0 +1,50 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/pathwayStats.R +\name{pathwayStats} +\alias{pathwayStats} +\title{Pathway Level Statistics} +\usage{ +pathwayStats(PGs, T, minSize=5, method="Stouffer") +} +\arguments{ +\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} + +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or +phosphorylation site that belong to the same pathway or kinase.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} +} +\value{ +a doublet corresponding to the enrichment after integration across all genes or substrates +that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +or kinase. +} +\description{ +Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +and calculates a combined statistics for those that belong to the same pathway or kinase. +} +\examples{ +# load the example data +data(PM) + +# load pathway annotations +data(Pathways) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) + +# compute statistics for all reactome pathways +gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +} + diff --git a/source/directPA_1.2/man/rotate2d.Rd b/source/directPA_1.2/man/rotate2d.Rd new file mode 100644 index 0000000..940baa2 --- /dev/null +++ b/source/directPA_1.2/man/rotate2d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate2d.R +\name{rotate2d} +\alias{rotate2d} +\title{Polar Coordinates Rotation} +\usage{ +rotate2d(T, degree = 0) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +} +\examples{ +# load the phosphoproteomics dataset +data(HEK) + +# convert statistics into z-scores +HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) + +# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +} + diff --git a/source/directPA_1.2/man/rotate3d.Rd b/source/directPA_1.2/man/rotate3d.Rd new file mode 100644 index 0000000..8367f4c --- /dev/null +++ b/source/directPA_1.2/man/rotate3d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate3d.R +\name{rotate3d} +\alias{rotate3d} +\title{Spherical Coordinates Rotation} +\usage{ +rotate3d(T, contrast) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +} + From cef13711f4172f778b3b3f3e38fd166933ba6807 Mon Sep 17 00:00:00 2001 From: PengyiYang Date: Tue, 17 Nov 2015 15:52:08 +1100 Subject: [PATCH 04/12] updates 17 Nov 2015 - 2 --- .Rbuildignore | 2 + .gitignore | 3 + DESCRIPTION | 14 ++ MD5 | 30 ++++ NAMESPACE | 11 ++ R/directExplorer2d.R | 93 ++++++++++++ R/directPA.R | 139 ++++++++++++++++++ R/geneStats.R | 43 ++++++ R/pathwayStats.R | 63 ++++++++ R/perturbPlot2d.R | 67 +++++++++ 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version/directPA_1.1/R/geneStats.R | 43 ++++++ .../directPA_1.1/R/pathwayStats.R | 63 ++++++++ previous version/directPA_1.1/R/rotate2d.R | 26 ++++ previous version/directPA_1.1/R/rotate3d.R | 44 ++++++ previous version/directPA_1.1/data/HEK.rda | Bin 0 -> 31259 bytes previous version/directPA_1.1/data/PM.rda | Bin 0 -> 27357 bytes .../directPA_1.1/data/Pathways.rda | Bin 0 -> 86220 bytes .../directPA_1.1/data/PhosphoSite.rda | Bin 0 -> 72692 bytes previous version/directPA_1.1/man/HEK.Rd | 8 + previous version/directPA_1.1/man/PM.Rd | 8 + .../directPA_1.1/man/Pathways.KEGG.Rd | 7 + .../directPA_1.1/man/Pathways.reactome.Rd | 7 + .../directPA_1.1/man/PhosphoSite.human.Rd | 8 + .../directPA_1.1/man/PhosphoSite.mouse.Rd | 8 + .../directPA_1.1/man/directEnrichPlot2d.Rd | 38 +++++ .../directPA_1.1/man/directExplorer2d.Rd | 59 ++++++++ .../directPA_1.1/man/directPA-package.Rd | 29 ++++ previous version/directPA_1.1/man/directPA.Rd | 92 ++++++++++++ .../directPA_1.1/man/geneStats.Rd | 36 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create mode 100644 source/directPA_1.2/man/pathwayStats.Rd create mode 100644 source/directPA_1.2/man/rotate2d.Rd create mode 100644 source/directPA_1.2/man/rotate3d.Rd diff --git a/.Rbuildignore b/.Rbuildignore new file mode 100644 index 0000000..91114bf --- /dev/null +++ b/.Rbuildignore @@ -0,0 +1,2 @@ +^.*\.Rproj$ +^\.Rproj\.user$ diff --git a/.gitignore b/.gitignore new file mode 100644 index 0000000..807ea25 --- /dev/null +++ b/.gitignore @@ -0,0 +1,3 @@ +.Rproj.user +.Rhistory +.RData diff --git a/DESCRIPTION b/DESCRIPTION new file mode 100644 index 0000000..c66409c --- /dev/null +++ b/DESCRIPTION @@ -0,0 +1,14 @@ +Package: directPA +Type: Package +Title: Direction Analysis for Pathways and Kinases +Version: 1.2 +Date: 2015-06-18 +Author: Pengyi Yang & Ellis Patrick +Maintainer: Pengyi Yang +Description: Direction analysis is a set of tools designed to integrate multiple experimental treatments for analyzing pathways and kinases from microarray, RNA-seq, proteomics, or phosphoproteomics data. +License: GPL-3 +Depends: R (>= 2.10.0), rgl, calibrate +Packaged: 2015-06-18 13:08:32 UTC; Pengyi +NeedsCompilation: no +Repository: CRAN +Date/Publication: 2015-06-18 18:00:22 diff --git a/MD5 b/MD5 new file mode 100644 index 0000000..3c8c935 --- /dev/null +++ b/MD5 @@ -0,0 +1,30 @@ +e05721202d78707adb25b5dadb9a905c *DESCRIPTION +32f7448fafd8f272e98f8f3053cc3f51 *NAMESPACE +61911534dedf36247467a549fbf57ec1 *R/directEnrichPlot2d.R +76cad031899e402b3ab2c8ee3bad588f *R/directExplorer2d.R +c193001c14dbc241fd64be4f11202254 *R/directPA.R +016bae5eb003dc005c4dd37ccc50f58e *R/geneStats.R +c94800bbd93527de53b09fc6bbce19a3 *R/pathwayStats.R +0fb8763a7e2602a1004522f451bbea85 *R/rotate2d.R +989f12754c30a855e238dd4a160a9806 *R/rotate3d.R +83648a7a500619337f5f19913ad14f71 *data/HEK.rda +079cc541e6cfd73ded8ab5b20a8687e6 *data/PM.rda +d8b015f9de0c68ca738fa61b8604add3 *data/Pathways.rda +a4829f119c5d601a1663ff68b6f033c9 *data/PhosphoELM.rda +48bf3aaf03017c2519d62984ea9587a2 *data/PhosphoSite.rda +a8abd01dded8fe74481dfc5d6ee6284a *man/HEK.Rd +c02262b58e37b1fefd695441707b8809 *man/PM.Rd +19d4e0276d10cd41f6d577ed5245ea12 *man/Pathways.KEGG.Rd +ffd89f58f9af47d0ff556011a5c3521c *man/Pathways.reactome.Rd +50f5b7321b62385a4af7e5c1ee1ee901 *man/PhosphoELM.human.Rd +8e9a9b800cbdb9e5e9764143c54ef6fb *man/PhosphoELM.mouse.Rd +9224f661163899dbf13b3b3a8775a167 *man/PhosphoSite.human.Rd +02df716ef54fb50fdecd8648c6883235 *man/PhosphoSite.mouse.Rd +3d1ba4de7361b2947991bf8b6f1de90b *man/directEnrichPlot2d.Rd +29b9f292ce686fa8368ab99a2a527a74 *man/directExplorer2d.Rd +8809fdbeedb03dd0c8672be14ef11383 *man/directPA-package.Rd +02069d7456d6905cc361baab683de7f8 *man/directPA.Rd +fcc77f5e75cdbce83340ec66f2777f47 *man/geneStats.Rd +aefc1a7e9de81ead1d8f8d1911002812 *man/pathwayStats.Rd +3e13753f1c19b7fb239f048db2376b20 *man/rotate2d.Rd +574ba8d8e806479f12b078b52b6de344 *man/rotate3d.Rd diff --git a/NAMESPACE b/NAMESPACE new file mode 100644 index 0000000..4cdaa62 --- /dev/null +++ b/NAMESPACE @@ -0,0 +1,11 @@ +# Generated by roxygen2 (4.1.0): do not edit by hand + +export(perturbPlot2d) +export(directExplorer2d) +export(directPA) +export(geneStats) +export(pathwayStats) +export(rotate2d) +export(rotate3d) +import(rgl) +import(calibrate) diff --git a/R/directExplorer2d.R b/R/directExplorer2d.R new file mode 100644 index 0000000..0d8e08b --- /dev/null +++ b/R/directExplorer2d.R @@ -0,0 +1,93 @@ +#' Batch Direction Analysis in 2-dimentional space +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", +#' path.method="Stouffer", top=10, ...) +#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns +#' are treatments vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param top the number of entries to be highlighted in the plot. +#' @param ... parameters for controlling the plot. +#' @return The the list of enrichment analysis in tables. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # test enrichment on 8 directions in polar coordinate system. +#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) +#' +#' # the direction are denoted as follow for the two treatments vs control: +#' # ++: up-regulated in both treatments +#' # +*: up-regulated in the first treatment and unchanged in the second treatment +#' # +-: up-regulated in the first treatment and down-regulated in the second treatment +#' # *-: unchanged in the first treatment and down-regulated in the second treatment +#' # --: down-regulated in both treatments +#' # -*: down-regulated in the first treatment and unchanged in the second treatment +#' # -+: down-regulated in the first treatment and up-regulated in the second treatment +#' # *+: unchanged in the first treatment and up-regulated in the second treatment +#' +#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both +#' # treatments (i.e. "--") and displa the top-10 entries +#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] +#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] +#' +directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { + + directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') + gene.tab <- matrix(NA, nrow(Tc), 8) + colnames(gene.tab) <- directionCode + rownames(gene.tab) <- rownames(Tc) + + path.tab <- NULL + if (!is.null(annotation)) { + path.tab <- matrix(NA, length(annotation), 9) + colnames(path.tab) <- c("size", directionCode) + rownames(path.tab) <- names(annotation) + } + + for(i in 0:7){ + # gene or phosphorylation site level + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + gene.tab[,(i+1)] <- gene.pvalues + + # pathway or kinase level + if (!is.null(annotation)) { + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + if (i == 0) { + path.tab[,1] <- unlist(gst[,"size"]) + } + path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) + } + } + + # highlight candidates + plot(Tc, col="gray", pch=16, ...) + abline(h=0, v=0, col="gold", lty=2) + abline(a=0, b=1, lty = 2, col="darkgreen") + color <- rainbow(8) + for (i in 1:8) { + ids <- names(sort(gene.tab[,i])[1:top]) + points(Tc[ids,], col=color[i], pch=16) + textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) + } + + results <- list() + results$gene.tab <- gene.tab + results$path.tab <- path.tab + return(results) +} + diff --git a/R/directPA.R b/R/directPA.R new file mode 100644 index 0000000..0e2b4e4 --- /dev/null +++ b/R/directPA.R @@ -0,0 +1,139 @@ +#' Direction Analysis for Pathways and Kinases +#' +#' The main function of direction Analysis. This function takes in a matrix of test statistics with +#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for +#' enrichment of pathways or kinases on the specified direction. +#' +#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +#' path.method="Stouffer", visualize=TRUE) +#' +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are +#' treatments vs control statistics. +#' @param direction the direction to be tested for enrichment. Either specified as a degree for +#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the +#' @return a list of enrichment for pathways or kinases +#' @export +#' @examples +#' +#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +#' # (180 degree to original direction) +#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +#' kst1[order(unlist(kst1[,1])),][1:20,] +#' +#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +#' # (135 degree to the original direction) +#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +#' kst2[order(unlist(kst2[,1])),][1:20,] +#' +#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +#' # (225 degree to the original direction) +#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +#' kst3[order(unlist(kst3[,1])),][1:20,] +#' +#' +#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +#' # load the proteomics dataset +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # display reactome pathways. Could be replaced by any other pathway databases +#' Pathways.reactome[1:5] +#' +#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +#' # on the original direction. +#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +#' gst1[order(unlist(gst1[,1])),][1:20,] +#' +#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +#' gst2[order(unlist(gst2[,1])),][1:20,] +#' +#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +#' gst3[order(unlist(gst3[,1])),][1:20,] +#' +directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE, ...){ + ## spherical coordinates for three-dimensional rotation + if (length(direction) == 3) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate3d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot3d(Tc, col=HC[rank(gene.pvalues)], size=5, ...) + abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) + abclines3d(x=0, a=direction, col="pink", lwd=5) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } + + ## polar coordinates for two-dimensional rotation + if (length(direction) == 1) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate2d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot(Tc, col=HC[rank(gene.pvalues)], pch=16, ...) + abline(v = 0,h = 0, lty=2, col="gold") + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } +} diff --git a/R/geneStats.R b/R/geneStats.R new file mode 100644 index 0000000..2d296dc --- /dev/null +++ b/R/geneStats.R @@ -0,0 +1,43 @@ +#' Gene Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +#' and calculates a combined statistics accross multiple treatments. +#' +#' @usage geneStats(T, method="OSP") +#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +#' comparison. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. +#' @return a p-value after integration across treatments. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +geneStats <- function(T, method="OSP") { + pvalue <- 0 + + if (method == "Stouffer") { + pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) + } else if (method == "OSP") { + p <- pnorm(T, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(T, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(T), lower.tail = FALSE) + } + + return (pvalue) +} diff --git a/R/pathwayStats.R b/R/pathwayStats.R new file mode 100644 index 0000000..107581f --- /dev/null +++ b/R/pathwayStats.R @@ -0,0 +1,63 @@ +#' Pathway Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +#' and calculates a combined statistics for those that belong to the same pathway or kinase. +#' +#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") +#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. +#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or +#' phosphorylation site that belong to the same pathway or kinase. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. +#' @return a doublet corresponding to the enrichment after integration across all genes or substrates +#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +#' or kinase. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +#' # compute statistics for all reactome pathways +#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +#' +pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ + pvalue <- 0 + Z <- T[names(T) %in% PGs] + + if (length(Z) >= minSize) { + if (method == "Stouffer") { + pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) + } else if (method == "OSP") { + p <- pnorm(Z, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(Z, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(Z), lower.tail = FALSE) + } + } else { + pvalue <- NA + } + + result <- list() + result$pvalue <- pvalue + result$size <- length(Z) + return (result) +} diff --git a/R/perturbPlot2d.R b/R/perturbPlot2d.R new file mode 100644 index 0000000..b46dcfb --- /dev/null +++ b/R/perturbPlot2d.R @@ -0,0 +1,67 @@ +#' Perturbation Plot +#' +#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the +#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment +#' of pathways or kinases in direction specific manner. +#' +#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments +#' vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or +#' substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param ... parameters for controling the plot. +#' @return a list of coordinates for pathways or kinases +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' perturbPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3) +#' +perturbPlot2d <- function(Tc, annotation, minSize=5, ...) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. filter the groups that are smaller than the minimun cutoff + DE = lapply(annotation, function(x){ + if(sum(rownames(Tc.zscores) %in% x) >= minSize) { + X <- Tc.zscores[rownames(Tc.zscores)%in%x,] + n = nrow(X) + Z1 = sum(X[,1])/sqrt(n) + Z2 = sum(X[,2])/sqrt(n) + list(Z1=Z1, Z2=Z2) + } + }) + + # step3. filter DE that has 0 element + DE <- DE[which(sapply(DE, length) != 0)] + Z1 <- unlist(sapply(DE, function(x){x[1]})) + Z2 <- unlist(sapply(DE, function(x){x[2]})) + + # visualization + plot(Z1, Z2, col="darkblue", pch=16, xlab=colnames(Tc)[1], ylab=colnames(Tc)[2], ...) + textxy(Z1,Z2, names(DE), col="black", cex=1) + abline(v=0, h=0, col="gold", lty=2) + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + + r <- ceiling(max(sqrt(Z1^2 + Z2^2))) + for(i in seq(0, r, r/5)) { + theta = seq(-3.14,3.14,0.05) + lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") + } + + points(Z1, Z2, col="darkblue", pch=16, cex=2) + + ## return the results + result <- list() + result$Z1 <- Z1 + result$Z2 <- Z2 + return(result) +} diff --git a/R/rotate2d.R b/R/rotate2d.R new file mode 100644 index 0000000..c6012ca --- /dev/null +++ b/R/rotate2d.R @@ -0,0 +1,26 @@ +#' Polar Coordinates Rotation +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage rotate2d(T, degree = 0) +#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # convert statistics into z-scores +#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) +#' +#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +#' +rotate2d = function(T, degree = 0){ + theta = atan2(T[,2], T[,1]) + degree + r = sqrt(T[,1]^2 + T[,2]^2) + x2 = r*cos(theta) + y2 = r*sin(theta) + return(cbind(x2, y2)) +} diff --git a/R/rotate3d.R b/R/rotate3d.R new file mode 100644 index 0000000..10f8a79 --- /dev/null +++ b/R/rotate3d.R @@ -0,0 +1,44 @@ +#' Spherical Coordinates Rotation +#' +#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +#' @usage rotate3d(T, contrast) +#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +rotate3d = function(T, contrast = c(1,1,1)){ + + a = contrast + b= c(1,1,1) # the original direction in contrast form is c(1,1,1) + + if(sum(a == b)==3)return(T) + if(sum(a == -b)==3)return(-T) + + a = a/sqrt(sum(a^2)) + b = b/sqrt(sum(b^2)) + u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) + u = u/sqrt(sum(u^2)) + R = matrix(0,3,3) + the = acos(sum(a*b)) + R[1,1] = cos(the) + u[1]^2*(1-cos(the)) + R[2,2] = cos(the) + u[2]^2*(1-cos(the)) + R[3,3] = cos(the) + u[3]^2*(1-cos(the)) + R[1,2] = u[1]*u[2]*(1-cos(the)) - u[3]*sin(the) + R[1,3] = u[1]*u[3]*(1-cos(the)) + u[2]*sin(the) + R[2,1] = u[2]*u[1]*(1-cos(the)) + u[3]*sin(the) + R[2,3] = u[2]*u[3]*(1-cos(the)) - u[1]*sin(the) + R[3,1] = u[3]*u[1]*(1-cos(the)) - u[2]*sin(the) + R[3,2] = u[3]*u[2]*(1-cos(the)) + u[1]*sin(the) + return(t(R%*%t(T))) +} diff --git a/data/HEK.rda b/data/HEK.rda new file mode 100644 index 0000000000000000000000000000000000000000..1e91f81e7a0d334aaf608b3085d9da5c3a727411 GIT binary patch literal 31259 zcmW)HiC>c0|9xdeWlCkGrh=u$^r@t>G8cGUDzj2%avEz)v8-&FWL#1~7EMh}4VP)m zT+pnXriIKVGZoy?TyQBhOHo0=Emm0{;K%p(`~mlMpVz(jywCfb^M+@xS@!?G)ue_~ zS(m%;9Q^aN^Iyga{$`c-tlw9?v?uGiyYOh!wdsnGmoHylzAA4RGr?t9T8Wz9wufYg z@1{3QUcFl0^F90U$~7Cx)>VF(ADalbP7s?`cQ%nfI{6#ES;`oIb~^H z{EDe(qO{4BJ|hg}8$)rsWw9X;P8InuyaBAfL`P%lKb0md-JTa#>isw#+aTI1`5=eV z+1{d&2@BGgfOd}g0~^1ErZ((?yhH}U8vU0SabdeD3f6Nh%~HK23mf`jzJQqX;($hO~JqvdmPY4VV-%EB7icb0Xq?naq@5ZETj6?`H}-L>lSe#0MZ{^(f={OFVc 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zXXF`M+VnX|D-u%7=J8d=op487T9sFu8=%!{h^LnIrbLM)w?ominYr7KsC_vbo=fN4 zHi#fl&v)Mpq5GB;wNZ;hjh8u2_4+Vn%~DMiipZ&b4{2N%s~nO&hTd}%ag5;XMT3cv zOyx)+Po4~B=4p>#uOF-CjObeL-gNlG0b6Y?x4D7Q;NJVATq7yp#Olv|Cp$32tbR5M*AOU>Iap!gYL)N_6Oce$8}DY>jE#0$egd8h()CXp7<#x7 zJ;MLWMCb3nl+QXu^j>yt@l4wlD-KN*`u zzNBXx(L7_lt87iX zTsA0PW-^WwN4R1ixCJC7cY_}*%;igRvvLh|K0&q-xO2&}3JAazLJ_OIWH~>ZRdI0M z!KVIs`>{Zs2moe1^&wVwt?jx)UYg;;;NNX!tIPB0d?wZ``LQDC;{dP!cBtGvx}Bce zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, +David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} diff --git a/man/directPA.Rd b/man/directPA.Rd new file mode 100644 index 0000000..50ffbeb --- /dev/null +++ b/man/directPA.Rd @@ -0,0 +1,93 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/directPA.R +\name{directPA} +\alias{directPA} +\title{Direction Analysis for Pathways and Kinases} +\usage{ +directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +path.method="Stouffer", visualize=TRUE) +} +\arguments{ +\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are +treatments vs control statistics.} + +\item{direction}{the direction to be tested for enrichment. Either specified as a degree for +two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} + +\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to +genes or substrates belong to each pathway or kinase, respectively.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{gene.method}{the method to be used for integrating statistics across treatments for each gene +or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is OSP.} + +\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation +sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is Stouffer.} + +\item{visualize}{whether to visualize the} +} +\value{ +a list of enrichment for pathways or kinases +} +\description{ +The main function of direction Analysis. This function takes in a matrix of test statistics with +two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +the annotation list such as pathway annotation or kinase-substrate annotation, and test for +enrichment of pathways or kinases on the specified direction. +} +\examples{ +## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +# load the phosphoproteomics dataset +data(HEK) + +# load the kinase-substrate annoations +data(PhosphoSite) + +# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +# (180 degree to original direction) +kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +kst1[order(unlist(kst1[,1])),][1:20,] + +# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +# (135 degree to the original direction) +kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +kst2[order(unlist(kst2[,1])),][1:20,] + +# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +# (225 degree to the original direction) +kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +kst3[order(unlist(kst3[,1])),][1:20,] + + +## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +# load the proteomics dataset +data(PM) + +# load pathway annotations +data(Pathways) + +# display reactome pathways. Could be replaced by any other pathway databases +Pathways.reactome[1:5] + +# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +# on the original direction. +gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +gst1[order(unlist(gst1[,1])),][1:20,] + +# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +gst2[order(unlist(gst2[,1])),][1:20,] + +# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +gst3[order(unlist(gst3[,1])),][1:20,] +} + diff --git a/man/geneStats.Rd b/man/geneStats.Rd new file mode 100644 index 0000000..455e3ef --- /dev/null +++ b/man/geneStats.Rd @@ -0,0 +1,36 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/geneStats.R +\name{geneStats} +\alias{geneStats} +\title{Gene Level Statistics} +\usage{ +geneStats(T, method="OSP") +} +\arguments{ +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +comparison.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} +} +\value{ +a p-value after integration across treatments. +} +\description{ +Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +and calculates a combined statistics accross multiple treatments. +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) +} + diff --git a/man/pathwayStats.Rd b/man/pathwayStats.Rd new file mode 100644 index 0000000..fec4ea5 --- /dev/null +++ b/man/pathwayStats.Rd @@ -0,0 +1,50 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/pathwayStats.R +\name{pathwayStats} +\alias{pathwayStats} +\title{Pathway Level Statistics} +\usage{ +pathwayStats(PGs, T, minSize=5, method="Stouffer") +} +\arguments{ +\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} + +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or +phosphorylation site that belong to the same pathway or kinase.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} +} +\value{ +a doublet corresponding to the enrichment after integration across all genes or substrates +that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +or kinase. +} +\description{ +Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +and calculates a combined statistics for those that belong to the same pathway or kinase. +} +\examples{ +# load the example data +data(PM) + +# load pathway annotations +data(Pathways) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) + +# compute statistics for all reactome pathways +gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +} + diff --git a/man/rotate2d.Rd b/man/rotate2d.Rd new file mode 100644 index 0000000..940baa2 --- /dev/null +++ b/man/rotate2d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate2d.R +\name{rotate2d} +\alias{rotate2d} +\title{Polar Coordinates Rotation} +\usage{ +rotate2d(T, degree = 0) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +} +\examples{ +# load the phosphoproteomics dataset +data(HEK) + +# convert statistics into z-scores +HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) + +# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +} + diff --git a/man/rotate3d.Rd b/man/rotate3d.Rd new file mode 100644 index 0000000..8367f4c --- /dev/null +++ b/man/rotate3d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate3d.R +\name{rotate3d} +\alias{rotate3d} +\title{Spherical Coordinates Rotation} +\usage{ +rotate3d(T, contrast) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +} + diff --git a/source/directPA_1.0/DESCRIPTION b/previous version/directPA_1.0/DESCRIPTION similarity index 100% rename from source/directPA_1.0/DESCRIPTION rename to previous version/directPA_1.0/DESCRIPTION diff --git a/source/directPA_1.0/MD5 b/previous version/directPA_1.0/MD5 similarity index 100% rename from source/directPA_1.0/MD5 rename to previous version/directPA_1.0/MD5 diff --git a/source/directPA_1.0/NAMESPACE b/previous version/directPA_1.0/NAMESPACE similarity index 100% rename from source/directPA_1.0/NAMESPACE rename to previous version/directPA_1.0/NAMESPACE diff --git a/source/directPA_1.0/R/directPA.R b/previous version/directPA_1.0/R/directPA.R similarity index 100% rename from source/directPA_1.0/R/directPA.R rename to previous version/directPA_1.0/R/directPA.R diff --git a/previous version/directPA_1.0/R/geneStats.R b/previous version/directPA_1.0/R/geneStats.R new file mode 100644 index 0000000..958b67a --- /dev/null +++ b/previous version/directPA_1.0/R/geneStats.R @@ -0,0 +1,20 @@ +# gene level statistics is calculated from here +# the default method for integration gene level information is OSP + +geneStats <- function(T, method="OSP") { + pvalue <- 0 + + if (method == "Stouffer") { + pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) + } else if (method == "OSP") { + p <- pnorm(T, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(T, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(T), lower.tail = FALSE) + } + + return (pvalue) +} diff --git a/source/directPA_1.0/R/pathwayStats.R b/previous version/directPA_1.0/R/pathwayStats.R similarity index 100% rename from source/directPA_1.0/R/pathwayStats.R rename to previous version/directPA_1.0/R/pathwayStats.R diff --git a/previous version/directPA_1.0/R/rotate2Sphere.R b/previous version/directPA_1.0/R/rotate2Sphere.R new file mode 100644 index 0000000..36fa4a3 --- /dev/null +++ b/previous version/directPA_1.0/R/rotate2Sphere.R @@ -0,0 +1,12 @@ +# polar coordinates rotation + +rotate2Sphere = function(T, degree = 0){ + # rotate to the direction that you want the test to face ie pi/4 degree + # T is a matrix of test statistics where the rows correspond to genes and the columns treatments + + theta = atan2(T[,2], T[,1]) + degree + r = sqrt(T[,1]^2 + T[,2]^2) + x2 = r*cos(theta) + y2 = r*sin(theta) + return(cbind(x2, y2)) +} diff --git a/previous version/directPA_1.0/R/rotate3Sphere.R b/previous version/directPA_1.0/R/rotate3Sphere.R new file mode 100644 index 0000000..e62422b --- /dev/null +++ b/previous version/directPA_1.0/R/rotate3Sphere.R @@ -0,0 +1,30 @@ +# spherical coordinates rotation + +rotate3Sphere = function(T, direction = c(1,1,1)){ + + # rotate to the direction that you want the test to face ie c(1,-1,-1) + # T is a matrix of test statistics where the rows correspond to genes and the columns treatments + + a = direction + b= c(1,1,1) # the original direction is c(1,1,1) + + if(sum(a == b)==3)return(T) + if(sum(a == -b)==3)return(-T) + + a = a/sqrt(sum(a^2)) + b = b/sqrt(sum(b^2)) + u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) + u = u/sqrt(sum(u^2)) + R = matrix(0,3,3) + the = acos(sum(a*b)) + R[1,1] = cos(the) + u[1]^2*(1-cos(the)) + R[2,2] = cos(the) + u[2]^2*(1-cos(the)) + R[3,3] = cos(the) + u[3]^2*(1-cos(the)) + R[1,2] = u[1]*u[2]*(1-cos(the)) - u[3]*sin(the) + R[1,3] = u[1]*u[3]*(1-cos(the)) + u[2]*sin(the) + R[2,1] = u[2]*u[1]*(1-cos(the)) + u[3]*sin(the) + R[2,3] = u[2]*u[3]*(1-cos(the)) - u[1]*sin(the) + R[3,1] = u[3]*u[1]*(1-cos(the)) - u[2]*sin(the) + R[3,2] = u[3]*u[2]*(1-cos(the)) + u[1]*sin(the) + return(t(R%*%t(T))) +} diff --git a/previous version/directPA_1.0/data/PM.RData b/previous version/directPA_1.0/data/PM.RData new file mode 100644 index 0000000000000000000000000000000000000000..3d713c684503fa0c1364214c8b2b01a629271232 GIT binary patch literal 27758 zcmV(~K+nG)iwFP!000002JM{nTU1^5@CRmS5HLXuEL23LL&bH58A=8i8Bjt|Y{dXY z48X!x48#r$R1{GVmDGJGK}1rzyJ2AHe8>B_?>WcspYUG%qU*!%T6^tv=2|vPdre7u zNeYD`Oc9pl|4ZeIG-3WfLKGTBg8x6&eE%W--zV^;Y#3|mu-##{1@(=ke%v;3Ehf4p z;v)B^4dmavsr~t0KdO&uy>xF`8{!@7;*9PMMV)(Z&2KZ?g$YZ0RDY2cM-f-@6CWfW zg^tAb9fsFkpk4WZ@6o=C(D&)`(Chdkh&M$pa(1T}CKNE=Z)ANxF1|vkWlet&YS))9 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+\alias{Ins} +\title{Insulin perturbation vesus control} +\description{ +The data object contains all quantified proteins with insulin perturbation vesus control in 3T3L1 cell lines. +} diff --git a/previous version/directPA_1.0/man/MK.Rd b/previous version/directPA_1.0/man/MK.Rd new file mode 100644 index 0000000..21bf9d2 --- /dev/null +++ b/previous version/directPA_1.0/man/MK.Rd @@ -0,0 +1,7 @@ +\name{MK} +\docType{data} +\alias{MK} +\title{MK perturbation vesus control} +\description{ +The data object contains all quantified proteins with MK perturbation vesus control in 3T3L1 cell lines. +} diff --git a/previous version/directPA_1.0/man/Wmn.Rd b/previous version/directPA_1.0/man/Wmn.Rd new file mode 100644 index 0000000..153ac6f --- /dev/null +++ b/previous version/directPA_1.0/man/Wmn.Rd @@ -0,0 +1,7 @@ +\name{Wmn} +\docType{data} +\alias{Wmn} +\title{Wortmannin perturbation vesus control} +\description{ +The data object contains all quantified proteins with Wortmannin perturbation vesus control in 3T3L1 cell lines. +} diff --git a/previous version/directPA_1.0/man/directPA-package.Rd b/previous version/directPA_1.0/man/directPA-package.Rd new file mode 100644 index 0000000..f24a827 --- /dev/null +++ b/previous version/directPA_1.0/man/directPA-package.Rd @@ -0,0 +1,29 @@ +\name{directPA-package} +\alias{directPA-package} +\docType{package} +\title{ +Direction Pathway Analysis Package +} +\description{ +The directPA-package is designed for analysing pathways in experiments with multiple perturbations. The combination effects of +different treatments are tested by rotating polar coordinates in two-dimentional space when the experiment contains two perturbations +and corresponding controls, or spherical coordinates in three-dimensional space when the experiment contains three perturbations and +corresponding controls. +} +\details{ +\tabular{ll}{ +Package: \tab directPA\cr +Type: \tab Package\cr +Version: \tab 1.0\cr +Date: \tab 2013-10-21\cr +License: \tab GPL-3\cr +} + +} +\author{ +Pengyi Yang & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, +David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} diff --git a/previous version/directPA_1.0/man/directPA.Rd b/previous version/directPA_1.0/man/directPA.Rd new file mode 100644 index 0000000..1cf4153 --- /dev/null +++ b/previous version/directPA_1.0/man/directPA.Rd @@ -0,0 +1,90 @@ +\name{directPA} +\alias{directPA} +\title{ +Direction Pathway Analysis +} +\description{ +The main function of direction pathway analysis package. +} +\usage{ +directPA(Tc, direction, pathway.list, minSize = 5, gene.method = "OSP", +path.method = "Stouffer", visualize = FALSE) +} +\arguments{ + \item{Tc}{ +The matrix of test statistics where rows correspond to proteins/genes and columns correspond to perturbations compared to controls. +} + \item{direction}{ +The direction to be tested. For two-dimensional rotation (experiments with two perturbations), directions are specified +as degree. For three-dimensional rotation (experiments with three perturbations), directions are specified as contrast. +See examples below for more details. +} + \item{pathway.list}{ +A pathway database in list format. See example below for details. +} + \item{minSize}{ +The minimum size of the pathway to be included in the report. +} + \item{gene.method}{ +A p-value integration method for protein/gene level combination. That is integrating information for each protein/gene accross +all perturbations (i.e. for each row of matrix Tc, integrating accross the columns). Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is OSP. +} + \item{path.method}{ +A p-value integration method for pathway level combination. That is integrating information of protein/gene included in a pathway, given +a pathway database. Available methods are Stouffer, OSP, Fisher, and maxP. Default method is Stouffer. +} + \item{visualize}{ +A boolean value indicating whether to visualize the protein/gene level integration statistics in a scatter plot. +} +} +\value{ +Return a matrix of test statistics for all given pathway that passed the minimum size cutoff on a specified test direction. +} +\author{ +Pengyi Yang & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} + +\examples{ + +# Load the example dataset. +data(PM) + +# Load the reactome pathway. +data(reactome) + +# Display reactome pathways. Could be replaced by any other pathway databases. +reactome.list + +# Combine test statistics of each perturbation vs control into a matrix of test statistics. +Tc = cbind(Ins,Wmn,MK) + +## Direction pathway analysis in two-dimensional space. Implemented as rotating by degree. +# (1) Testing combined effect of Ins and Wmn vs controls on original direction. +gst1 <- directPA(Tc[,c(1,2)], direction=0, pathway.list=reactome.list) +# Display the top 20 pathways ranked by integrated p-value. +gst1[order(unlist(gst1[,1])),][1:20,] + +# (2) Testing combined effect of Ins and MK vs controls on the pi/2 direction. +gst2 <- directPA(Tc[,c(1,3)], direction=pi/2, pathway.list=reactome.list) +gst2[order(unlist(gst2[,1])),][1:20,] + +## Direction pathway analysis in three-dimensional space. Implemnted as rotating by contrast. +# (1) Testing combined effect of all 3 perturbations vs controls on the original direction. +gst3 <- directPA(Tc, direction=c(1,1,1), pathway.list=reactome.list) +gst3[order(unlist(gst3[,1])),][1:20,] + +# (2) Testing combined effect of all 3 perturbations vs controls on direction c(1,-1, 0). +# This rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree +gst4 <- directPA(Tc, direction=c(1,-1,0), pathway.list=reactome.list) +gst4[order(unlist(gst4[,1])),][1:20,] + +# (3) Testing combined effect of all 3 perturbations vs controls on direction c(1,-1, 1). +# This rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree +gst5 <- directPA(Tc, direction=c(1,-1,1), pathway.list=reactome.list) +gst5[order(unlist(gst5[,1])),][1:20,] + +} diff --git a/previous version/directPA_1.0/man/geneStats.Rd b/previous version/directPA_1.0/man/geneStats.Rd new file mode 100644 index 0000000..cd70c0a --- /dev/null +++ b/previous version/directPA_1.0/man/geneStats.Rd @@ -0,0 +1,64 @@ +\name{geneStats} +\alias{geneStats} +\title{ +Gene Level Statistics +} +\description{ +Calculating protein/gene level statistics accross multiple perturbations. +} +\usage{ +geneStats(T, method = "OSP") +} +%- maybe also 'usage' for other objects documented here. +\arguments{ + \item{T}{ +A matrix of test statistics (normally after rotation), where rows correspond to proteins/genes and columns correspond to perturbations. +} + \item{method}{ +The p-value integration method for combining accross multiple perturbations. +Available methods are Stouffer, OSP, Fisher, and maxP. The default method is OSP. +} +} +\value{ +Return the integration p-value for a protein/gene. +} +\author{ +Pengyi Yang & Ellis Patrick +} + + +\examples{ + +# Load the example data. +data(PM) + + +## (1) For three perturbatins vesus controls, use three dimentional rotation. +Tc <- cbind(Ins, Wmn, MK) + +# Rotate the test statistics matrix to the direction of interest. +Tc.rotated <- rotate3Sphere(Tc, direction=c(1,-1,0)) + +# Integrate protein/gene statistics accross perturmations. +gene.pvalues <- apply(Tc.rotated, 1, geneStats) + +# Visualize the rotation statistics. +library(rgl) +HC <- rainbow(length(gene.pvalues)*1.2) +plot3d(Tc, col=HC[rank(gene.pvalues)], size=5) +abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) +abclines3d(x=0, a=c(1,-1,0), col="pink", lwd=5) + + +## (2) For two perturbations vesus controls, use two dimentional rotation. +Tc <- cbind(Ins, Wmn) +Tc.rotated <- rotate2Sphere(Tc, degree=pi/4) +gene.pvalues <- apply(Tc.rotated, 1, geneStats) + +# Visualize the rotation statistics. +HC = rainbow(length(gene.pvalues)*1.2) +plot(Tc, col=HC[rank(gene.pvalues)], pch=16) +abline(v = 0,h = 0, lty=2, col="gold") +} + + diff --git a/previous version/directPA_1.0/man/pathwayStats.Rd b/previous version/directPA_1.0/man/pathwayStats.Rd new file mode 100644 index 0000000..2b6a8eb --- /dev/null +++ b/previous version/directPA_1.0/man/pathwayStats.Rd @@ -0,0 +1,56 @@ +\name{pathwayStats} +\alias{pathwayStats} +\title{ +Pathway Level Statistics +} +\description{ +Calculating pathway level statistics by combining protein/gene belong to this pathway. +} +\usage{ +pathwayStats(PGs, T, minSize, method = "Stouffer") +} +%- maybe also 'usage' for other objects documented here. +\arguments{ + \item{PGs}{ +An array of names indicating proteins/genes belong to a given pathway. +} + \item{T}{ +An array of test statistics of all proteins/genes. This should be the zsore transformed output of geneStats. +} + \item{minSize}{ +The minimum size of a pathway to be reported. +} + \item{method}{ +A p-value integration method for combining proteins/genes in a given pathway. +} +} +\value{ + \item{pvalue }{Integrated pvalue of the given pathway} + \item{size }{size of the given pathway} +} +\author{ +Pengyi Yang & Ellis Patrick +} + +\examples{ + +# Load the example data. +data(PM) +Tc = cbind(Ins, Wmn, MK) + +# Load reactome pathway. +data(reactome) + +# Rotate the test statistics matrix to the direction of interest. +Tc.rotated <- rotate3Sphere(Tc, direction=c(1, -1, 0)) + +# Integrate protein/gene statistics accross perturmations. +gene.pvalues <- apply(Tc.rotated, 1, geneStats) + +# Transform the pvlaue computed from geneStats to zsocres. +gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + +# Compute statistics for all reactome pathway +gst <- t(sapply(reactome.list, pathwayStats, gene.zscores, minSize=5)) + +} diff --git a/previous version/directPA_1.0/man/reactome.list.Rd b/previous version/directPA_1.0/man/reactome.list.Rd new file mode 100644 index 0000000..3cfe4e7 --- /dev/null +++ b/previous version/directPA_1.0/man/reactome.list.Rd @@ -0,0 +1,10 @@ +\name{reactome.list} +\docType{data} +\alias{reactome.list} +\title{Reactome Database} +\description{ +Reactome database in list format. Each list element contains an array of gene names corresponding to the genes contains in a pathway. The name of the list element is the pathway name. +} +\references{ +Joshi-Tope, G., et al. Reactome: a knowledgebase of biological pathways. Nucleic Acids Research 33.suppl 1 (2005): D428-D432. +} diff --git a/previous version/directPA_1.0/man/rotate2Sphere.Rd b/previous version/directPA_1.0/man/rotate2Sphere.Rd new file mode 100644 index 0000000..11b7dee --- /dev/null +++ b/previous version/directPA_1.0/man/rotate2Sphere.Rd @@ -0,0 +1,39 @@ +\name{rotate2Sphere} +\alias{rotate2Sphere} +\title{ +Polar Coordinates Rotation +} +\description{ +Rotate to the direction of interest in polar coordinates by degree (i.e pi/4). +} +\usage{ +rotate2Sphere(T, degree = 0) +} +\arguments{ + \item{T}{ +A matrix of test statistics. The rows correspond to protein/genes and the columns correspond to perturbations. The number of colmuns should be 2. +} + \item{degree}{ +The degree to be rotated. +} +} +\value{ +A transformed matrix with respect to the direction of interest. +} +\author{ +Pengyi Yang & Ellis Patrick +} + +\examples{ +# Load the example dataset. +data(PM) + +# Combine two perturbations into a single matrix. +# The matrix contains test statistics of each protein w.r.t each perturbation. +Tc <- cbind(Ins, Wmn) + +# Rotate the matrix by pi/2 in polar coordinates. +Tc.rotated <- rotate2Sphere(Tc, degree = pi/2) +} + + diff --git a/previous version/directPA_1.0/man/rotate3Sphere.Rd b/previous version/directPA_1.0/man/rotate3Sphere.Rd new file mode 100644 index 0000000..93b28dd --- /dev/null +++ b/previous version/directPA_1.0/man/rotate3Sphere.Rd @@ -0,0 +1,37 @@ +\name{rotate3Sphere} +\alias{rotate3Sphere} +\title{ +Spherical Coordinates Rotation +} +\description{ +Rotate to the direction of interest in spherical coordinates by contrast (i.e 1, -1, -1). +} +\usage{ +rotate3Sphere(T, direction = c(1, 1, 1)) +} +\arguments{ + \item{T}{ +A matrix of test statistics. The rows correspond to protein/genes and the columns correspond to perturbations. The number of columns should be 3. +} + \item{direction}{ +The angle to be rotated in contrast format (see example below). +} +} +\value{ +A transformed matrix with respect to the direction of interest. +} +\author{ +Pengyi Yang & Ellis Patrick +} + +\examples{ +# Load the example data. +data(PM) + +# Combine three perturbations into a single matrix. +# The matrix contains test statistics of each protein w.r.t each perturbations. +Tc <- cbind(Ins, Wmn, MK) + +# Rotate the matrix by contrast 1, -1, -1 in spherical coordinates. +Tc.rotated <- rotate3Sphere(Tc, direction = c(1, -1, -1)) +} diff --git a/previous version/directPA_1.1/DESCRIPTION b/previous version/directPA_1.1/DESCRIPTION new file mode 100644 index 0000000..698fb4f --- /dev/null +++ b/previous version/directPA_1.1/DESCRIPTION @@ -0,0 +1,10 @@ +Package: directPA +Type: Package +Title: Direction Pathway Analysis +Version: 1.1 +Date: 2015-03-13 +Author: Pengyi Yang & Ellis Patrick +Maintainer: Pengyi Yang +Description: A package for pathway or kinase analysis in experiments with multiple treatment designs. +License: GPL-3 +Depends: R (>= 2.10.0), rgl, calibrate diff --git a/previous version/directPA_1.1/NAMESPACE b/previous version/directPA_1.1/NAMESPACE new file mode 100644 index 0000000..7fe39f5 --- /dev/null +++ b/previous version/directPA_1.1/NAMESPACE @@ -0,0 +1,11 @@ +# Generated by roxygen2 (4.1.0): do not edit by hand + +export(directEnrichPlot2d) +export(directExplorer2d) +export(directPA) +export(geneStats) +export(pathwayStats) +export(rotate2d) +export(rotate3d) +import(rgl) +import(calibrate) diff --git a/previous version/directPA_1.1/R/directEnrichPlot2d.R b/previous version/directPA_1.1/R/directEnrichPlot2d.R new file mode 100644 index 0000000..44b660a --- /dev/null +++ b/previous version/directPA_1.1/R/directEnrichPlot2d.R @@ -0,0 +1,67 @@ +#' Direction Enrichment Plot for Pathways and Kinases +#' +#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the +#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment +#' of pathways or kinases in direction specific manner. +#' +#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments +#' vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or +#' substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param ... parameters for controling the plot. +#' @return a list of coordinates for pathways or kinases +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' directEnrichPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3, xlim=c(-8, 3), ylim=c(-6, 3)) +#' +directEnrichPlot2d <- function(Tc, annotation, minSize=5, ...) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. filter the groups that are smaller than the minimun cutoff + DE = lapply(annotation, function(x){ + if(sum(rownames(Tc.zscores) %in% x) >= minSize) { + X <- Tc.zscores[rownames(Tc.zscores)%in%x,] + n = nrow(X) + Z1 = sum(X[,1])/sqrt(n) + Z2 = sum(X[,2])/sqrt(n) + list(Z1=Z1, Z2=Z2) + } + }) + + # step3. filter DE that has 0 element + DE <- DE[which(sapply(DE, length) != 0)] + Z1 <- unlist(sapply(DE, function(x){x[1]})) + Z2 <- unlist(sapply(DE, function(x){x[2]})) + + # visualization + plot(Z1, Z2, col="darkblue", pch=16, ...) + textxy(Z1,Z2, names(DE), col="black", cex=1) + abline(v=0, h=0, col="gold", lty=2) + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + + r <- ceiling(max(sqrt(Z1^2 + Z2^2))) + for(i in seq(0, r, r/5)) { + theta = seq(-3.14,3.14,0.05) + lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") + } + + points(Z1, Z2, col="darkblue", pch=16, cex=2) + + ## return the results + result <- list() + result$Z1 <- Z1 + result$Z2 <- Z2 + return(result) +} diff --git a/previous version/directPA_1.1/R/directExplorer2d.R b/previous version/directPA_1.1/R/directExplorer2d.R new file mode 100644 index 0000000..41135d9 --- /dev/null +++ b/previous version/directPA_1.1/R/directExplorer2d.R @@ -0,0 +1,92 @@ +#' Batch Direction Analysis in 2-dimentional space +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) +#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns +#' are treatments vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param top the number of entries to be highlighted in the plot. +#' @param ... parameters for controlling the plot. +#' @return The the list of enrichment analysis in tables. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # test enrichment on 8 directions in polar coordinate system. +#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) +#' +#' # the direction are denoted as follow for the two treatments vs control: +#' # ++: up-regulated in both treatments +#' # +*: up-regulated in the first treatment and unchanged in the second treatment +#' # +-: up-regulated in the first treatment and down-regulated in the second treatment +#' # *-: unchanged in the first treatment and down-regulated in the second treatment +#' # --: down-regulated in both treatments +#' # -*: down-regulated in the first treatment and unchanged in the second treatment +#' # -+: down-regulated in the first treatment and up-regulated in the second treatment +#' # *+: unchanged in the first treatment and up-regulated in the second treatment +#' +#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both +#' # treatments (i.e. "--") and displa the top-10 entries +#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] +#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] +#' +directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { + + directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') + gene.tab <- matrix(NA, nrow(Tc), 8) + colnames(gene.tab) <- directionCode + rownames(gene.tab) <- rownames(Tc) + + path.tab <- NULL + if (!is.null(annotation)) { + path.tab <- matrix(NA, length(annotation), 9) + colnames(path.tab) <- c("size", directionCode) + rownames(path.tab) <- names(annotation) + } + + for(i in 0:7){ + # gene or phosphorylation site level + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + gene.tab[,(i+1)] <- gene.pvalues + + # pathway or kinase level + if (!is.null(annotation)) { + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + if (i == 0) { + path.tab[,1] <- unlist(gst[,"size"]) + } + path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) + } + } + + # highlight candidates + plot(Tc, col="gray", pch=16, ...) + abline(h=0, v=0, col="gold", lty=2) + abline(a=0, b=1, lty = 2, col="darkgreen") + color <- rainbow(8) + for (i in 1:8) { + ids <- names(sort(gene.tab[,i])[1:top]) + points(Tc[ids,], col=color[i], pch=16) + textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) + } + + results <- list() + results$gene.tab <- gene.tab + results$path.tab <- path.tab + return(results) +} + diff --git a/previous version/directPA_1.1/R/directPA.R b/previous version/directPA_1.1/R/directPA.R new file mode 100644 index 0000000..b365e57 --- /dev/null +++ b/previous version/directPA_1.1/R/directPA.R @@ -0,0 +1,128 @@ +#' Direction Analysis for Pathways and Kinases +#' +#' The main function of direction Analysis. This function takes in a matrix of test statistics with +#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for +#' enrichment of pathways or kinases on the specified direction. +#' +#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE) +#' +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are +#' treatments vs control statistics. +#' @param direction the direction to be tested for enrichment. Either specified as a degree for +#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the +#' @return a list of enrichment for pathways or kinases +#' @export +#' @examples +#' +#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +#' # (180 degree to original direction) +#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +#' kst1[order(unlist(kst1[,1])),][1:20,] +#' +#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +#' # (135 degree to the original direction) +#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +#' kst2[order(unlist(kst2[,1])),][1:20,] +#' +#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +#' # (225 degree to the original direction) +#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +#' kst3[order(unlist(kst3[,1])),][1:20,] +#' +#' +#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +#' # load the proteomics dataset +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # display reactome pathways. Could be replaced by any other pathway databases +#' Pathways.reactome[1:5] +#' +#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +#' # on the original direction. +#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +#' gst1[order(unlist(gst1[,1])),][1:20,] +#' +#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +#' gst2[order(unlist(gst2[,1])),][1:20,] +#' +#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +#' gst3[order(unlist(gst3[,1])),][1:20,] +#' +directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE){ + ## spherical coordinates for three-dimensional rotation + if (length(direction) == 3) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate3d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot3d(Tc, col=HC[rank(gene.pvalues)], size=5) + abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) + abclines3d(x=0, a=direction, col="pink", lwd=5) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + return(gst) + } + + ## polar coordinates for two-dimensional rotation + if (length(direction) == 1) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate2d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot(Tc, col=HC[rank(gene.pvalues)], pch=16) + abline(v = 0,h = 0, lty=2, col="gold") + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + return(gst) + } +} diff --git a/previous version/directPA_1.1/R/geneStats.R b/previous version/directPA_1.1/R/geneStats.R new file mode 100644 index 0000000..2d296dc --- /dev/null +++ b/previous version/directPA_1.1/R/geneStats.R @@ -0,0 +1,43 @@ +#' Gene Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +#' and calculates a combined statistics accross multiple treatments. +#' +#' @usage geneStats(T, method="OSP") +#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +#' comparison. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. +#' @return a p-value after integration across treatments. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +geneStats <- function(T, method="OSP") { + pvalue <- 0 + + if (method == "Stouffer") { + pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) + } else if (method == "OSP") { + p <- pnorm(T, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(T, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(T), lower.tail = FALSE) + } + + return (pvalue) +} diff --git a/previous version/directPA_1.1/R/pathwayStats.R b/previous version/directPA_1.1/R/pathwayStats.R new file mode 100644 index 0000000..0b6f177 --- /dev/null +++ b/previous version/directPA_1.1/R/pathwayStats.R @@ -0,0 +1,63 @@ +#' Pathway Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +#' and calculates a combined statistics for those that belong to the same pathway or kinase. +#' +#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") +#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. +#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or +#' phosphorylation site that belong to the same pathway or kinase. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. +#' @return a doublet corresponding to the enrichment after integration across all genes or substrates +#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +#' or kinase. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +#' # compute statistics for all reactome pathways +#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +#' +pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ + pvalue <- 0 + Z <- T[names(T) %in% PGs] + + if (length(Z) >= minSize) { + if (method == "Stouffer") { + pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) + } else if (method == "OSP") { + p <- pnorm(Z, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(Z, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(Z), lower.tail = FALSE) + } + } else { + pvalue <- NA + } + + result <- list() + result$pvalue <- pvalue + result$size <- length(Z) + return (result) +} diff --git a/previous version/directPA_1.1/R/rotate2d.R b/previous version/directPA_1.1/R/rotate2d.R new file mode 100644 index 0000000..c6012ca --- /dev/null +++ b/previous version/directPA_1.1/R/rotate2d.R @@ -0,0 +1,26 @@ +#' Polar Coordinates Rotation +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage rotate2d(T, degree = 0) +#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # convert statistics into z-scores +#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) +#' +#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +#' +rotate2d = function(T, degree = 0){ + theta = atan2(T[,2], T[,1]) + degree + r = sqrt(T[,1]^2 + T[,2]^2) + x2 = r*cos(theta) + y2 = r*sin(theta) + return(cbind(x2, y2)) +} diff --git a/previous version/directPA_1.1/R/rotate3d.R b/previous version/directPA_1.1/R/rotate3d.R new file mode 100644 index 0000000..10f8a79 --- /dev/null +++ b/previous version/directPA_1.1/R/rotate3d.R @@ -0,0 +1,44 @@ +#' Spherical Coordinates Rotation +#' +#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +#' @usage rotate3d(T, contrast) +#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +rotate3d = function(T, contrast = c(1,1,1)){ + + a = contrast + b= c(1,1,1) # the original direction in contrast form is c(1,1,1) + + if(sum(a == b)==3)return(T) + if(sum(a == -b)==3)return(-T) + + a = a/sqrt(sum(a^2)) + b = b/sqrt(sum(b^2)) + u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) + u = u/sqrt(sum(u^2)) + R = matrix(0,3,3) + the = acos(sum(a*b)) + R[1,1] = cos(the) + u[1]^2*(1-cos(the)) + R[2,2] = cos(the) + u[2]^2*(1-cos(the)) + R[3,3] = cos(the) + u[3]^2*(1-cos(the)) + 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zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, +David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} diff --git a/previous version/directPA_1.1/man/directPA.Rd b/previous version/directPA_1.1/man/directPA.Rd new file mode 100644 index 0000000..6fba82e --- /dev/null +++ b/previous version/directPA_1.1/man/directPA.Rd @@ -0,0 +1,92 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/directPA.R +\name{directPA} +\alias{directPA} +\title{Direction Analysis for Pathways and Kinases} +\usage{ +directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE) +} +\arguments{ +\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are +treatments vs control statistics.} + +\item{direction}{the direction to be tested for enrichment. Either specified as a degree for +two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} + +\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to +genes or substrates belong to each pathway or kinase, respectively.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{gene.method}{the method to be used for integrating statistics across treatments for each gene +or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is OSP.} + +\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation +sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is Stouffer.} + +\item{visualize}{whether to visualize the} +} +\value{ +a list of enrichment for pathways or kinases +} +\description{ +The main function of direction Analysis. This function takes in a matrix of test statistics with +two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +the annotation list such as pathway annotation or kinase-substrate annotation, and test for +enrichment of pathways or kinases on the specified direction. +} +\examples{ +## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +# load the phosphoproteomics dataset +data(HEK) + +# load the kinase-substrate annoations +data(PhosphoSite) + +# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +# (180 degree to original direction) +kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +kst1[order(unlist(kst1[,1])),][1:20,] + +# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +# (135 degree to the original direction) +kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +kst2[order(unlist(kst2[,1])),][1:20,] + +# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +# (225 degree to the original direction) +kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +kst3[order(unlist(kst3[,1])),][1:20,] + + +## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +# load the proteomics dataset +data(PM) + +# load pathway annotations +data(Pathways) + +# display reactome pathways. Could be replaced by any other pathway databases +Pathways.reactome[1:5] + +# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +# on the original direction. +gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +gst1[order(unlist(gst1[,1])),][1:20,] + +# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +gst2[order(unlist(gst2[,1])),][1:20,] + +# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +gst3[order(unlist(gst3[,1])),][1:20,] +} + diff --git a/previous version/directPA_1.1/man/geneStats.Rd b/previous version/directPA_1.1/man/geneStats.Rd new file mode 100644 index 0000000..455e3ef --- /dev/null +++ b/previous version/directPA_1.1/man/geneStats.Rd @@ -0,0 +1,36 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/geneStats.R +\name{geneStats} +\alias{geneStats} +\title{Gene Level Statistics} +\usage{ +geneStats(T, method="OSP") +} +\arguments{ +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +comparison.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} +} +\value{ +a p-value after integration across treatments. +} +\description{ +Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +and calculates a combined statistics accross multiple treatments. +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) +} + diff --git a/previous version/directPA_1.1/man/pathwayStats.Rd b/previous version/directPA_1.1/man/pathwayStats.Rd new file mode 100644 index 0000000..fec4ea5 --- /dev/null +++ b/previous version/directPA_1.1/man/pathwayStats.Rd @@ -0,0 +1,50 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/pathwayStats.R +\name{pathwayStats} +\alias{pathwayStats} +\title{Pathway Level Statistics} +\usage{ +pathwayStats(PGs, T, minSize=5, method="Stouffer") +} +\arguments{ +\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} + +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or +phosphorylation site that belong to the same pathway or kinase.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} +} +\value{ +a doublet corresponding to the enrichment after integration across all genes or substrates +that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +or kinase. +} +\description{ +Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +and calculates a combined statistics for those that belong to the same pathway or kinase. +} +\examples{ +# load the example data +data(PM) + +# load pathway annotations +data(Pathways) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) + +# compute statistics for all reactome pathways +gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +} + diff --git a/previous version/directPA_1.1/man/rotate2d.Rd b/previous version/directPA_1.1/man/rotate2d.Rd new file mode 100644 index 0000000..940baa2 --- /dev/null +++ b/previous version/directPA_1.1/man/rotate2d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate2d.R +\name{rotate2d} +\alias{rotate2d} +\title{Polar Coordinates Rotation} +\usage{ +rotate2d(T, degree = 0) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +} +\examples{ +# load the phosphoproteomics dataset +data(HEK) + +# convert statistics into z-scores +HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) + +# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +} + diff --git a/previous version/directPA_1.1/man/rotate3d.Rd b/previous version/directPA_1.1/man/rotate3d.Rd new file mode 100644 index 0000000..8367f4c --- /dev/null +++ b/previous version/directPA_1.1/man/rotate3d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate3d.R +\name{rotate3d} +\alias{rotate3d} +\title{Spherical Coordinates Rotation} +\usage{ +rotate3d(T, contrast) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +} + diff --git a/source/directPA_1.2/.Rbuildignore b/source/directPA_1.2/.Rbuildignore new file mode 100644 index 0000000..91114bf --- /dev/null +++ b/source/directPA_1.2/.Rbuildignore @@ -0,0 +1,2 @@ +^.*\.Rproj$ +^\.Rproj\.user$ diff --git a/source/directPA_1.2/.gitignore b/source/directPA_1.2/.gitignore new file mode 100644 index 0000000..807ea25 --- /dev/null +++ b/source/directPA_1.2/.gitignore @@ -0,0 +1,3 @@ +.Rproj.user +.Rhistory +.RData diff --git a/source/directPA_1.2/DESCRIPTION b/source/directPA_1.2/DESCRIPTION new file mode 100644 index 0000000..c66409c --- /dev/null +++ b/source/directPA_1.2/DESCRIPTION @@ -0,0 +1,14 @@ +Package: directPA +Type: Package +Title: Direction Analysis for Pathways and Kinases +Version: 1.2 +Date: 2015-06-18 +Author: Pengyi Yang & Ellis Patrick +Maintainer: Pengyi Yang +Description: Direction analysis is a set of tools designed to integrate multiple experimental treatments for analyzing pathways and kinases from microarray, RNA-seq, proteomics, or phosphoproteomics data. +License: GPL-3 +Depends: R (>= 2.10.0), rgl, calibrate +Packaged: 2015-06-18 13:08:32 UTC; Pengyi +NeedsCompilation: no +Repository: CRAN +Date/Publication: 2015-06-18 18:00:22 diff --git a/source/directPA_1.2/MD5 b/source/directPA_1.2/MD5 new file mode 100644 index 0000000..3c8c935 --- /dev/null +++ b/source/directPA_1.2/MD5 @@ -0,0 +1,30 @@ +e05721202d78707adb25b5dadb9a905c *DESCRIPTION +32f7448fafd8f272e98f8f3053cc3f51 *NAMESPACE +61911534dedf36247467a549fbf57ec1 *R/directEnrichPlot2d.R +76cad031899e402b3ab2c8ee3bad588f *R/directExplorer2d.R +c193001c14dbc241fd64be4f11202254 *R/directPA.R +016bae5eb003dc005c4dd37ccc50f58e *R/geneStats.R +c94800bbd93527de53b09fc6bbce19a3 *R/pathwayStats.R +0fb8763a7e2602a1004522f451bbea85 *R/rotate2d.R +989f12754c30a855e238dd4a160a9806 *R/rotate3d.R +83648a7a500619337f5f19913ad14f71 *data/HEK.rda +079cc541e6cfd73ded8ab5b20a8687e6 *data/PM.rda +d8b015f9de0c68ca738fa61b8604add3 *data/Pathways.rda +a4829f119c5d601a1663ff68b6f033c9 *data/PhosphoELM.rda +48bf3aaf03017c2519d62984ea9587a2 *data/PhosphoSite.rda +a8abd01dded8fe74481dfc5d6ee6284a *man/HEK.Rd +c02262b58e37b1fefd695441707b8809 *man/PM.Rd +19d4e0276d10cd41f6d577ed5245ea12 *man/Pathways.KEGG.Rd +ffd89f58f9af47d0ff556011a5c3521c *man/Pathways.reactome.Rd +50f5b7321b62385a4af7e5c1ee1ee901 *man/PhosphoELM.human.Rd +8e9a9b800cbdb9e5e9764143c54ef6fb *man/PhosphoELM.mouse.Rd +9224f661163899dbf13b3b3a8775a167 *man/PhosphoSite.human.Rd +02df716ef54fb50fdecd8648c6883235 *man/PhosphoSite.mouse.Rd +3d1ba4de7361b2947991bf8b6f1de90b *man/directEnrichPlot2d.Rd +29b9f292ce686fa8368ab99a2a527a74 *man/directExplorer2d.Rd +8809fdbeedb03dd0c8672be14ef11383 *man/directPA-package.Rd +02069d7456d6905cc361baab683de7f8 *man/directPA.Rd +fcc77f5e75cdbce83340ec66f2777f47 *man/geneStats.Rd +aefc1a7e9de81ead1d8f8d1911002812 *man/pathwayStats.Rd +3e13753f1c19b7fb239f048db2376b20 *man/rotate2d.Rd +574ba8d8e806479f12b078b52b6de344 *man/rotate3d.Rd diff --git a/source/directPA_1.2/NAMESPACE b/source/directPA_1.2/NAMESPACE new file mode 100644 index 0000000..4cdaa62 --- /dev/null +++ b/source/directPA_1.2/NAMESPACE @@ -0,0 +1,11 @@ +# Generated by roxygen2 (4.1.0): do not edit by hand + +export(perturbPlot2d) +export(directExplorer2d) +export(directPA) +export(geneStats) +export(pathwayStats) +export(rotate2d) +export(rotate3d) +import(rgl) +import(calibrate) diff --git a/source/directPA_1.2/R/directExplorer2d.R b/source/directPA_1.2/R/directExplorer2d.R new file mode 100644 index 0000000..0d8e08b --- /dev/null +++ b/source/directPA_1.2/R/directExplorer2d.R @@ -0,0 +1,93 @@ +#' Batch Direction Analysis in 2-dimentional space +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", +#' path.method="Stouffer", top=10, ...) +#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns +#' are treatments vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param top the number of entries to be highlighted in the plot. +#' @param ... parameters for controlling the plot. +#' @return The the list of enrichment analysis in tables. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # test enrichment on 8 directions in polar coordinate system. +#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) +#' +#' # the direction are denoted as follow for the two treatments vs control: +#' # ++: up-regulated in both treatments +#' # +*: up-regulated in the first treatment and unchanged in the second treatment +#' # +-: up-regulated in the first treatment and down-regulated in the second treatment +#' # *-: unchanged in the first treatment and down-regulated in the second treatment +#' # --: down-regulated in both treatments +#' # -*: down-regulated in the first treatment and unchanged in the second treatment +#' # -+: down-regulated in the first treatment and up-regulated in the second treatment +#' # *+: unchanged in the first treatment and up-regulated in the second treatment +#' +#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both +#' # treatments (i.e. "--") and displa the top-10 entries +#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] +#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] +#' +directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { + + directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') + gene.tab <- matrix(NA, nrow(Tc), 8) + colnames(gene.tab) <- directionCode + rownames(gene.tab) <- rownames(Tc) + + path.tab <- NULL + if (!is.null(annotation)) { + path.tab <- matrix(NA, length(annotation), 9) + colnames(path.tab) <- c("size", directionCode) + rownames(path.tab) <- names(annotation) + } + + for(i in 0:7){ + # gene or phosphorylation site level + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + gene.tab[,(i+1)] <- gene.pvalues + + # pathway or kinase level + if (!is.null(annotation)) { + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + if (i == 0) { + path.tab[,1] <- unlist(gst[,"size"]) + } + path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) + } + } + + # highlight candidates + plot(Tc, col="gray", pch=16, ...) + abline(h=0, v=0, col="gold", lty=2) + abline(a=0, b=1, lty = 2, col="darkgreen") + color <- rainbow(8) + for (i in 1:8) { + ids <- names(sort(gene.tab[,i])[1:top]) + points(Tc[ids,], col=color[i], pch=16) + textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) + } + + results <- list() + results$gene.tab <- gene.tab + results$path.tab <- path.tab + return(results) +} + diff --git a/source/directPA_1.2/R/directPA.R b/source/directPA_1.2/R/directPA.R new file mode 100644 index 0000000..0e2b4e4 --- /dev/null +++ b/source/directPA_1.2/R/directPA.R @@ -0,0 +1,139 @@ +#' Direction Analysis for Pathways and Kinases +#' +#' The main function of direction Analysis. This function takes in a matrix of test statistics with +#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for +#' enrichment of pathways or kinases on the specified direction. +#' +#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +#' path.method="Stouffer", visualize=TRUE) +#' +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are +#' treatments vs control statistics. +#' @param direction the direction to be tested for enrichment. Either specified as a degree for +#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to +#' genes or substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene +#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation +#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the +#' @return a list of enrichment for pathways or kinases +#' @export +#' @examples +#' +#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +#' # (180 degree to original direction) +#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +#' kst1[order(unlist(kst1[,1])),][1:20,] +#' +#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +#' # (135 degree to the original direction) +#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +#' kst2[order(unlist(kst2[,1])),][1:20,] +#' +#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +#' # (225 degree to the original direction) +#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +#' kst3[order(unlist(kst3[,1])),][1:20,] +#' +#' +#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +#' # load the proteomics dataset +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # display reactome pathways. Could be replaced by any other pathway databases +#' Pathways.reactome[1:5] +#' +#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +#' # on the original direction. +#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +#' gst1[order(unlist(gst1[,1])),][1:20,] +#' +#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +#' gst2[order(unlist(gst2[,1])),][1:20,] +#' +#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +#' gst3[order(unlist(gst3[,1])),][1:20,] +#' +directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE, ...){ + ## spherical coordinates for three-dimensional rotation + if (length(direction) == 3) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate3d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot3d(Tc, col=HC[rank(gene.pvalues)], size=5, ...) + abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) + abclines3d(x=0, a=direction, col="pink", lwd=5) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } + + ## polar coordinates for two-dimensional rotation + if (length(direction) == 1) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. rotate z-scores + Tc.rotated <- rotate2d(Tc.zscores, direction) + + # step 3. integrate statistics across treatments + gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) + + if (visualize == TRUE) { + HC = rainbow(length(gene.pvalues)*1.2) + plot(Tc, col=HC[rank(gene.pvalues)], pch=16, ...) + abline(v = 0,h = 0, lty=2, col="gold") + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + } + + # step 4. integrate statistics for pathways or kinases + gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) + gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) + + result <- list() + result$gene.pvalues <- gene.pvalues + result$gst <- gst + return(result) + } +} diff --git a/source/directPA_1.2/R/geneStats.R b/source/directPA_1.2/R/geneStats.R new file mode 100644 index 0000000..2d296dc --- /dev/null +++ b/source/directPA_1.2/R/geneStats.R @@ -0,0 +1,43 @@ +#' Gene Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +#' and calculates a combined statistics accross multiple treatments. +#' +#' @usage geneStats(T, method="OSP") +#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +#' comparison. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. +#' @return a p-value after integration across treatments. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +geneStats <- function(T, method="OSP") { + pvalue <- 0 + + if (method == "Stouffer") { + pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) + } else if (method == "OSP") { + p <- pnorm(T, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(T, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(T), lower.tail = FALSE) + } + + return (pvalue) +} diff --git a/source/directPA_1.2/R/pathwayStats.R b/source/directPA_1.2/R/pathwayStats.R new file mode 100644 index 0000000..107581f --- /dev/null +++ b/source/directPA_1.2/R/pathwayStats.R @@ -0,0 +1,63 @@ +#' Pathway Level Statistics +#' +#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +#' and calculates a combined statistics for those that belong to the same pathway or kinase. +#' +#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") +#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. +#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or +#' phosphorylation site that belong to the same pathway or kinase. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param method the p-value integration method for combining accross multiple treatments. Available methods +#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. +#' @return a doublet corresponding to the enrichment after integration across all genes or substrates +#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +#' or kinase. +#' @export +#' +#' @examples +#' # load the example data +#' data(PM) +#' +#' # load pathway annotations +#' data(Pathways) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +#' # combine rotated statistics across treatments +#' gene.pvalues <- apply(PM.rotated, 1, geneStats) +#' +#' # compute statistics for all reactome pathways +#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +#' +pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ + pvalue <- 0 + Z <- T[names(T) %in% PGs] + + if (length(Z) >= minSize) { + if (method == "Stouffer") { + pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) + } else if (method == "OSP") { + p <- pnorm(Z, lower.tail = TRUE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) + } else if (method == "Fisher") { + p <- pnorm(Z, lower.tail = FALSE) + pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) + } else if (method == "maxP") { + pvalue <- pnorm(max(Z), lower.tail = FALSE) + } + } else { + pvalue <- NA + } + + result <- list() + result$pvalue <- pvalue + result$size <- length(Z) + return (result) +} diff --git a/source/directPA_1.2/R/perturbPlot2d.R b/source/directPA_1.2/R/perturbPlot2d.R new file mode 100644 index 0000000..b46dcfb --- /dev/null +++ b/source/directPA_1.2/R/perturbPlot2d.R @@ -0,0 +1,67 @@ +#' Perturbation Plot +#' +#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the +#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment +#' of pathways or kinases in direction specific manner. +#' +#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) +#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments +#' vs control statistics. +#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or +#' substrates belong to each pathway or kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param ... parameters for controling the plot. +#' @return a list of coordinates for pathways or kinases +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' perturbPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3) +#' +perturbPlot2d <- function(Tc, annotation, minSize=5, ...) { + + # step 1. convert statistics into z-scores + Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) + + # step 2. filter the groups that are smaller than the minimun cutoff + DE = lapply(annotation, function(x){ + if(sum(rownames(Tc.zscores) %in% x) >= minSize) { + X <- Tc.zscores[rownames(Tc.zscores)%in%x,] + n = nrow(X) + Z1 = sum(X[,1])/sqrt(n) + Z2 = sum(X[,2])/sqrt(n) + list(Z1=Z1, Z2=Z2) + } + }) + + # step3. filter DE that has 0 element + DE <- DE[which(sapply(DE, length) != 0)] + Z1 <- unlist(sapply(DE, function(x){x[1]})) + Z2 <- unlist(sapply(DE, function(x){x[2]})) + + # visualization + plot(Z1, Z2, col="darkblue", pch=16, xlab=colnames(Tc)[1], ylab=colnames(Tc)[2], ...) + textxy(Z1,Z2, names(DE), col="black", cex=1) + abline(v=0, h=0, col="gold", lty=2) + abline(a=0, b=1, col="darkgreen", lty=2) + abline(a=0, b=-1, col="darkgreen", lty=2) + + r <- ceiling(max(sqrt(Z1^2 + Z2^2))) + for(i in seq(0, r, r/5)) { + theta = seq(-3.14,3.14,0.05) + lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") + } + + points(Z1, Z2, col="darkblue", pch=16, cex=2) + + ## return the results + result <- list() + result$Z1 <- Z1 + result$Z2 <- Z2 + return(result) +} diff --git a/source/directPA_1.2/R/rotate2d.R b/source/directPA_1.2/R/rotate2d.R new file mode 100644 index 0000000..c6012ca --- /dev/null +++ b/source/directPA_1.2/R/rotate2d.R @@ -0,0 +1,26 @@ +#' Polar Coordinates Rotation +#' +#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +#' @usage rotate2d(T, degree = 0) +#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # convert statistics into z-scores +#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) +#' +#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +#' +rotate2d = function(T, degree = 0){ + theta = atan2(T[,2], T[,1]) + degree + r = sqrt(T[,1]^2 + T[,2]^2) + x2 = r*cos(theta) + y2 = r*sin(theta) + return(cbind(x2, y2)) +} diff --git a/source/directPA_1.2/R/rotate3d.R b/source/directPA_1.2/R/rotate3d.R new file mode 100644 index 0000000..10f8a79 --- /dev/null +++ b/source/directPA_1.2/R/rotate3d.R @@ -0,0 +1,44 @@ +#' Spherical Coordinates Rotation +#' +#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +#' @usage rotate3d(T, contrast) +#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +#' sites and the columns are treatments vs control statistics. +#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). +#' @return A rotated matrix with respect to the direction of interest. +#' @export +#' @examples +#' # load the example data +#' data(PM) +#' +#' # convert statistics into z-scores +#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) +#' +#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +#' +rotate3d = function(T, contrast = c(1,1,1)){ + + a = contrast + b= c(1,1,1) # the original direction in contrast form is c(1,1,1) + + if(sum(a == b)==3)return(T) + if(sum(a == -b)==3)return(-T) + + a = a/sqrt(sum(a^2)) + b = b/sqrt(sum(b^2)) + u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) + u = u/sqrt(sum(u^2)) + R = matrix(0,3,3) + the = acos(sum(a*b)) + R[1,1] = cos(the) + u[1]^2*(1-cos(the)) + R[2,2] = cos(the) + u[2]^2*(1-cos(the)) + R[3,3] = cos(the) + u[3]^2*(1-cos(the)) + 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zL80D?idv`n{h{jM%DBslXiOX7kg=a*-#ZID{S`)(-bM}+M0!(Ou~M2 zR~=WKI4MDJQ9XLeQ60yM*7&TsFcBuz&nA+#MBfD*|H&^y zr>Ftr|HnqFz%IZd7p-kJVq7m6hs{K8Dg+=^R>izt$LLxn1T zh@^;~SvrT$N^04HCUD=*W0t=v+z}}usOBWS^IJqi63kKZXE=Szv5 zXXF`M+VnX|D-u%7=J8d=op487T9sFu8=%!{h^LnIrbLM)w?ominYr7KsC_vbo=fN4 zHi#fl&v)Mpq5GB;wNZ;hjh8u2_4+Vn%~DMiipZ&b4{2N%s~nO&hTd}%ag5;XMT3cv zOyx)+Po4~B=4p>#uOF-CjObeL-gNlG0b6Y?x4D7Q;NJVATq7yp#Olv|Cp$32tbR5M*AOU>Iap!gYL)N_6Oce$8}DY>jE#0$egd8h()CXp7<#x7 zJ;MLWMCb3nl+QXu^j>yt@l4wlD-KN*`u zzNBXx(L7_lt87iX zTsA0PW-^WwN4R1ixCJC7cY_}*%;igRvvLh|K0&q-xO2&}3JAazLJ_OIWH~>ZRdI0M z!KVIs`>{Zs2moe1^&wVwt?jx)UYg;;;NNX!tIPB0d?wZ``LQDC;{dP!cBtGvx}Bce zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick +} +\references{ +Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, +David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. +} diff --git a/source/directPA_1.2/man/directPA.Rd b/source/directPA_1.2/man/directPA.Rd new file mode 100644 index 0000000..50ffbeb --- /dev/null +++ b/source/directPA_1.2/man/directPA.Rd @@ -0,0 +1,93 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/directPA.R +\name{directPA} +\alias{directPA} +\title{Direction Analysis for Pathways and Kinases} +\usage{ +directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", +path.method="Stouffer", visualize=TRUE) +} +\arguments{ +\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are +treatments vs control statistics.} + +\item{direction}{the direction to be tested for enrichment. Either specified as a degree for +two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} + +\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to +genes or substrates belong to each pathway or kinase, respectively.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{gene.method}{the method to be used for integrating statistics across treatments for each gene +or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is OSP.} + +\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation +sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +Default method is Stouffer.} + +\item{visualize}{whether to visualize the} +} +\value{ +a list of enrichment for pathways or kinases +} +\description{ +The main function of direction Analysis. This function takes in a matrix of test statistics with +two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and +the annotation list such as pathway annotation or kinase-substrate annotation, and test for +enrichment of pathways or kinases on the specified direction. +} +\examples{ +## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) +# load the phosphoproteomics dataset +data(HEK) + +# load the kinase-substrate annoations +data(PhosphoSite) + +# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +# (180 degree to original direction) +kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +kst1[order(unlist(kst1[,1])),][1:20,] + +# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +# (135 degree to the original direction) +kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +kst2[order(unlist(kst2[,1])),][1:20,] + +# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +# (225 degree to the original direction) +kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +kst3[order(unlist(kst3[,1])),][1:20,] + + +## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) +# load the proteomics dataset +data(PM) + +# load pathway annotations +data(Pathways) + +# display reactome pathways. Could be replaced by any other pathway databases +Pathways.reactome[1:5] + +# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast +# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) +# on the original direction. +gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +gst1[order(unlist(gst1[,1])),][1:20,] + +# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. +gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +gst2[order(unlist(gst2[,1])),][1:20,] + +# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) +# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. +gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +gst3[order(unlist(gst3[,1])),][1:20,] +} + diff --git a/source/directPA_1.2/man/geneStats.Rd b/source/directPA_1.2/man/geneStats.Rd new file mode 100644 index 0000000..455e3ef --- /dev/null +++ b/source/directPA_1.2/man/geneStats.Rd @@ -0,0 +1,36 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/geneStats.R +\name{geneStats} +\alias{geneStats} +\title{Gene Level Statistics} +\usage{ +geneStats(T, method="OSP") +} +\arguments{ +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control +comparison.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} +} +\value{ +a p-value after integration across treatments. +} +\description{ +Takes a vector of statistics with each element corresponds to a treatment vs control comparison, +and calculates a combined statistics accross multiple treatments. +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) +} + diff --git a/source/directPA_1.2/man/pathwayStats.Rd b/source/directPA_1.2/man/pathwayStats.Rd new file mode 100644 index 0000000..fec4ea5 --- /dev/null +++ b/source/directPA_1.2/man/pathwayStats.Rd @@ -0,0 +1,50 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/pathwayStats.R +\name{pathwayStats} +\alias{pathwayStats} +\title{Pathway Level Statistics} +\usage{ +pathwayStats(PGs, T, minSize=5, method="Stouffer") +} +\arguments{ +\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} + +\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or +phosphorylation site that belong to the same pathway or kinase.} + +\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups +that are smaller than the minSize will be removed from the analysis.} + +\item{method}{the p-value integration method for combining accross multiple treatments. Available methods +are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} +} +\value{ +a doublet corresponding to the enrichment after integration across all genes or substrates +that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway +or kinase. +} +\description{ +Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, +and calculates a combined statistics for those that belong to the same pathway or kinase. +} +\examples{ +# load the example data +data(PM) + +# load pathway annotations +data(Pathways) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) + +# combine rotated statistics across treatments +gene.pvalues <- apply(PM.rotated, 1, geneStats) + +# compute statistics for all reactome pathways +gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) +gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) +} + diff --git a/source/directPA_1.2/man/rotate2d.Rd b/source/directPA_1.2/man/rotate2d.Rd new file mode 100644 index 0000000..940baa2 --- /dev/null +++ b/source/directPA_1.2/man/rotate2d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate2d.R +\name{rotate2d} +\alias{rotate2d} +\title{Polar Coordinates Rotation} +\usage{ +rotate2d(T, degree = 0) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). +} +\examples{ +# load the phosphoproteomics dataset +data(HEK) + +# convert statistics into z-scores +HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) + +# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). +HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) +} + diff --git a/source/directPA_1.2/man/rotate3d.Rd b/source/directPA_1.2/man/rotate3d.Rd new file mode 100644 index 0000000..8367f4c --- /dev/null +++ b/source/directPA_1.2/man/rotate3d.Rd @@ -0,0 +1,31 @@ +% Generated by roxygen2 (4.1.0): do not edit by hand +% Please edit documentation in R/rotate3d.R +\name{rotate3d} +\alias{rotate3d} +\title{Spherical Coordinates Rotation} +\usage{ +rotate3d(T, contrast) +} +\arguments{ +\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation +sites and the columns are treatments vs control statistics.} + +\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} +} +\value{ +A rotated matrix with respect to the direction of interest. +} +\description{ +Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). +} +\examples{ +# load the example data +data(PM) + +# convert statistics into z-scores +PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) + +# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). +PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) +} + From d477dc1bd9a5127f220144a21b8cdcca62c00a39 Mon Sep 17 00:00:00 2001 From: PengyiYang Date: Tue, 17 Nov 2015 16:04:15 +1100 Subject: [PATCH 05/12] remove --- source/directPA_1.0/R/geneStats.R | 20 --- source/directPA_1.0/R/rotate2Sphere.R | 12 -- source/directPA_1.0/R/rotate3Sphere.R | 30 ---- source/directPA_1.0/data/PM.RData | Bin 27758 -> 0 bytes source/directPA_1.0/data/reactome.RData | Bin 63436 -> 0 bytes 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delete mode 100644 source/directPA_1.2/man/Pathways.reactome.Rd delete mode 100644 source/directPA_1.2/man/PhosphoELM.human.Rd delete mode 100644 source/directPA_1.2/man/PhosphoELM.mouse.Rd delete mode 100644 source/directPA_1.2/man/PhosphoSite.human.Rd delete mode 100644 source/directPA_1.2/man/PhosphoSite.mouse.Rd delete mode 100644 source/directPA_1.2/man/directEnrichPlot2d.Rd delete mode 100644 source/directPA_1.2/man/directExplorer2d.Rd delete mode 100644 source/directPA_1.2/man/directPA-package.Rd delete mode 100644 source/directPA_1.2/man/directPA.Rd delete mode 100644 source/directPA_1.2/man/geneStats.Rd delete mode 100644 source/directPA_1.2/man/pathwayStats.Rd delete mode 100644 source/directPA_1.2/man/rotate2d.Rd delete mode 100644 source/directPA_1.2/man/rotate3d.Rd diff --git a/source/directPA_1.0/R/geneStats.R b/source/directPA_1.0/R/geneStats.R deleted file mode 100644 index 958b67a..0000000 --- a/source/directPA_1.0/R/geneStats.R +++ /dev/null @@ -1,20 +0,0 @@ -# gene level statistics is calculated from here -# the default method for integration gene level information is OSP - -geneStats <- function(T, method="OSP") { - pvalue <- 0 - - if (method == "Stouffer") { - pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) - } else if (method == "OSP") { - p <- pnorm(T, lower.tail = TRUE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) - } else if (method == "Fisher") { - p <- pnorm(T, lower.tail = FALSE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) - } else if (method == "maxP") { - pvalue <- pnorm(max(T), lower.tail = FALSE) - } - - return (pvalue) -} diff --git a/source/directPA_1.0/R/rotate2Sphere.R b/source/directPA_1.0/R/rotate2Sphere.R deleted file mode 100644 index 36fa4a3..0000000 --- a/source/directPA_1.0/R/rotate2Sphere.R +++ /dev/null @@ -1,12 +0,0 @@ -# polar coordinates rotation - -rotate2Sphere = function(T, degree = 0){ - # rotate to the direction that you want the test to face ie pi/4 degree - # T is a matrix of test statistics where the rows correspond to genes and the columns treatments - - theta = atan2(T[,2], T[,1]) + degree - r = sqrt(T[,1]^2 + T[,2]^2) - x2 = r*cos(theta) - y2 = r*sin(theta) - return(cbind(x2, y2)) -} diff --git a/source/directPA_1.0/R/rotate3Sphere.R b/source/directPA_1.0/R/rotate3Sphere.R deleted file mode 100644 index e62422b..0000000 --- a/source/directPA_1.0/R/rotate3Sphere.R +++ /dev/null @@ -1,30 +0,0 @@ -# spherical coordinates rotation - -rotate3Sphere = function(T, direction = c(1,1,1)){ - - # rotate to the direction that you want the test to face ie c(1,-1,-1) - # T is a matrix of test statistics where the rows correspond to genes and the columns treatments - - a = direction - b= c(1,1,1) # the original direction is c(1,1,1) - - if(sum(a == b)==3)return(T) - if(sum(a == -b)==3)return(-T) - - a = a/sqrt(sum(a^2)) - b = b/sqrt(sum(b^2)) - u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) - u = 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control} -\description{ -The data object contains all quantified proteins with insulin perturbation vesus control in 3T3L1 cell lines. -} diff --git a/source/directPA_1.0/man/MK.Rd b/source/directPA_1.0/man/MK.Rd deleted file mode 100644 index 21bf9d2..0000000 --- a/source/directPA_1.0/man/MK.Rd +++ /dev/null @@ -1,7 +0,0 @@ -\name{MK} -\docType{data} -\alias{MK} -\title{MK perturbation vesus control} -\description{ -The data object contains all quantified proteins with MK perturbation vesus control in 3T3L1 cell lines. -} diff --git a/source/directPA_1.0/man/Wmn.Rd b/source/directPA_1.0/man/Wmn.Rd deleted file mode 100644 index 153ac6f..0000000 --- a/source/directPA_1.0/man/Wmn.Rd +++ /dev/null @@ -1,7 +0,0 @@ -\name{Wmn} -\docType{data} -\alias{Wmn} -\title{Wortmannin perturbation vesus control} -\description{ -The data object contains all quantified proteins with Wortmannin perturbation vesus control in 3T3L1 cell lines. -} diff --git a/source/directPA_1.0/man/directPA-package.Rd b/source/directPA_1.0/man/directPA-package.Rd deleted file mode 100644 index f24a827..0000000 --- a/source/directPA_1.0/man/directPA-package.Rd +++ /dev/null @@ -1,29 +0,0 @@ -\name{directPA-package} -\alias{directPA-package} -\docType{package} -\title{ -Direction Pathway Analysis Package -} -\description{ -The directPA-package is designed for analysing pathways in experiments with multiple perturbations. The combination effects of -different treatments are tested by rotating polar coordinates in two-dimentional space when the experiment contains two perturbations -and corresponding controls, or spherical coordinates in three-dimensional space when the experiment contains three perturbations and -corresponding controls. -} -\details{ -\tabular{ll}{ -Package: \tab directPA\cr -Type: \tab Package\cr -Version: \tab 1.0\cr -Date: \tab 2013-10-21\cr -License: \tab GPL-3\cr -} - -} -\author{ -Pengyi Yang & Ellis Patrick -} -\references{ -Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, -David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. -} diff --git a/source/directPA_1.0/man/directPA.Rd b/source/directPA_1.0/man/directPA.Rd deleted file mode 100644 index 1cf4153..0000000 --- a/source/directPA_1.0/man/directPA.Rd +++ /dev/null @@ -1,90 +0,0 @@ -\name{directPA} -\alias{directPA} -\title{ -Direction Pathway Analysis -} -\description{ -The main function of direction pathway analysis package. -} -\usage{ -directPA(Tc, direction, pathway.list, minSize = 5, gene.method = "OSP", -path.method = "Stouffer", visualize = FALSE) -} -\arguments{ - \item{Tc}{ -The matrix of test statistics where rows correspond to proteins/genes and columns correspond to perturbations compared to controls. -} - \item{direction}{ -The direction to be tested. For two-dimensional rotation (experiments with two perturbations), directions are specified -as degree. For three-dimensional rotation (experiments with three perturbations), directions are specified as contrast. -See examples below for more details. -} - \item{pathway.list}{ -A pathway database in list format. See example below for details. -} - \item{minSize}{ -The minimum size of the pathway to be included in the report. -} - \item{gene.method}{ -A p-value integration method for protein/gene level combination. That is integrating information for each protein/gene accross -all perturbations (i.e. for each row of matrix Tc, integrating accross the columns). Available methods are Stouffer, OSP, Fisher, and maxP. -Default method is OSP. -} - \item{path.method}{ -A p-value integration method for pathway level combination. That is integrating information of protein/gene included in a pathway, given -a pathway database. Available methods are Stouffer, OSP, Fisher, and maxP. Default method is Stouffer. -} - \item{visualize}{ -A boolean value indicating whether to visualize the protein/gene level integration statistics in a scatter plot. -} -} -\value{ -Return a matrix of test statistics for all given pathway that passed the minimum size cutoff on a specified test direction. -} -\author{ -Pengyi Yang & Ellis Patrick -} -\references{ -Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. -} - -\examples{ - -# Load the example dataset. -data(PM) - -# Load the reactome pathway. -data(reactome) - -# Display reactome pathways. Could be replaced by any other pathway databases. -reactome.list - -# Combine test statistics of each perturbation vs control into a matrix of test statistics. -Tc = cbind(Ins,Wmn,MK) - -## Direction pathway analysis in two-dimensional space. Implemented as rotating by degree. -# (1) Testing combined effect of Ins and Wmn vs controls on original direction. -gst1 <- directPA(Tc[,c(1,2)], direction=0, pathway.list=reactome.list) -# Display the top 20 pathways ranked by integrated p-value. -gst1[order(unlist(gst1[,1])),][1:20,] - -# (2) Testing combined effect of Ins and MK vs controls on the pi/2 direction. -gst2 <- directPA(Tc[,c(1,3)], direction=pi/2, pathway.list=reactome.list) -gst2[order(unlist(gst2[,1])),][1:20,] - -## Direction pathway analysis in three-dimensional space. Implemnted as rotating by contrast. -# (1) Testing combined effect of all 3 perturbations vs controls on the original direction. -gst3 <- directPA(Tc, direction=c(1,1,1), pathway.list=reactome.list) -gst3[order(unlist(gst3[,1])),][1:20,] - -# (2) Testing combined effect of all 3 perturbations vs controls on direction c(1,-1, 0). -# This rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree -gst4 <- directPA(Tc, direction=c(1,-1,0), pathway.list=reactome.list) -gst4[order(unlist(gst4[,1])),][1:20,] - -# (3) Testing combined effect of all 3 perturbations vs controls on direction c(1,-1, 1). -# This rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree -gst5 <- directPA(Tc, direction=c(1,-1,1), pathway.list=reactome.list) -gst5[order(unlist(gst5[,1])),][1:20,] - -} diff --git a/source/directPA_1.0/man/geneStats.Rd b/source/directPA_1.0/man/geneStats.Rd deleted file mode 100644 index cd70c0a..0000000 --- a/source/directPA_1.0/man/geneStats.Rd +++ /dev/null @@ -1,64 +0,0 @@ -\name{geneStats} -\alias{geneStats} -\title{ -Gene Level Statistics -} -\description{ -Calculating protein/gene level statistics accross multiple perturbations. -} -\usage{ -geneStats(T, method = "OSP") -} -%- maybe also 'usage' for other objects documented here. -\arguments{ - \item{T}{ -A matrix of test statistics (normally after rotation), where rows correspond to proteins/genes and columns correspond to perturbations. -} - \item{method}{ -The p-value integration method for combining accross multiple perturbations. -Available methods are Stouffer, OSP, Fisher, and maxP. The default method is OSP. -} -} -\value{ -Return the integration p-value for a protein/gene. -} -\author{ -Pengyi Yang & Ellis Patrick -} - - -\examples{ - -# Load the example data. -data(PM) - - -## (1) For three perturbatins vesus controls, use three dimentional rotation. -Tc <- cbind(Ins, Wmn, MK) - -# Rotate the test statistics matrix to the direction of interest. -Tc.rotated <- rotate3Sphere(Tc, direction=c(1,-1,0)) - -# Integrate protein/gene statistics accross perturmations. -gene.pvalues <- apply(Tc.rotated, 1, geneStats) - -# Visualize the rotation statistics. -library(rgl) -HC <- rainbow(length(gene.pvalues)*1.2) -plot3d(Tc, col=HC[rank(gene.pvalues)], size=5) -abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) -abclines3d(x=0, a=c(1,-1,0), col="pink", lwd=5) - - -## (2) For two perturbations vesus controls, use two dimentional rotation. -Tc <- cbind(Ins, Wmn) -Tc.rotated <- rotate2Sphere(Tc, degree=pi/4) -gene.pvalues <- apply(Tc.rotated, 1, geneStats) - -# Visualize the rotation statistics. -HC = rainbow(length(gene.pvalues)*1.2) -plot(Tc, col=HC[rank(gene.pvalues)], pch=16) -abline(v = 0,h = 0, lty=2, col="gold") -} - - diff --git a/source/directPA_1.0/man/pathwayStats.Rd b/source/directPA_1.0/man/pathwayStats.Rd deleted file mode 100644 index 2b6a8eb..0000000 --- a/source/directPA_1.0/man/pathwayStats.Rd +++ /dev/null @@ -1,56 +0,0 @@ -\name{pathwayStats} -\alias{pathwayStats} -\title{ -Pathway Level Statistics -} -\description{ -Calculating pathway level statistics by combining protein/gene belong to this pathway. -} -\usage{ -pathwayStats(PGs, T, minSize, method = "Stouffer") -} -%- maybe also 'usage' for other objects documented here. -\arguments{ - \item{PGs}{ -An array of names indicating proteins/genes belong to a given pathway. -} - \item{T}{ -An array of test statistics of all proteins/genes. This should be the zsore transformed output of geneStats. -} - \item{minSize}{ -The minimum size of a pathway to be reported. -} - \item{method}{ -A p-value integration method for combining proteins/genes in a given pathway. -} -} -\value{ - \item{pvalue }{Integrated pvalue of the given pathway} - \item{size }{size of the given pathway} -} -\author{ -Pengyi Yang & Ellis Patrick -} - -\examples{ - -# Load the example data. -data(PM) -Tc = cbind(Ins, Wmn, MK) - -# Load reactome pathway. -data(reactome) - -# Rotate the test statistics matrix to the direction of interest. -Tc.rotated <- rotate3Sphere(Tc, direction=c(1, -1, 0)) - -# Integrate protein/gene statistics accross perturmations. -gene.pvalues <- apply(Tc.rotated, 1, geneStats) - -# Transform the pvlaue computed from geneStats to zsocres. -gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - -# Compute statistics for all reactome pathway -gst <- t(sapply(reactome.list, pathwayStats, gene.zscores, minSize=5)) - -} diff --git a/source/directPA_1.0/man/reactome.list.Rd b/source/directPA_1.0/man/reactome.list.Rd deleted file mode 100644 index 3cfe4e7..0000000 --- a/source/directPA_1.0/man/reactome.list.Rd +++ /dev/null @@ -1,10 +0,0 @@ -\name{reactome.list} -\docType{data} -\alias{reactome.list} -\title{Reactome Database} -\description{ -Reactome database in list format. Each list element contains an array of gene names corresponding to the genes contains in a pathway. The name of the list element is the pathway name. -} -\references{ -Joshi-Tope, G., et al. Reactome: a knowledgebase of biological pathways. Nucleic Acids Research 33.suppl 1 (2005): D428-D432. -} diff --git a/source/directPA_1.0/man/rotate2Sphere.Rd b/source/directPA_1.0/man/rotate2Sphere.Rd deleted file mode 100644 index 11b7dee..0000000 --- a/source/directPA_1.0/man/rotate2Sphere.Rd +++ /dev/null @@ -1,39 +0,0 @@ -\name{rotate2Sphere} -\alias{rotate2Sphere} -\title{ -Polar Coordinates Rotation -} -\description{ -Rotate to the direction of interest in polar coordinates by degree (i.e pi/4). -} -\usage{ -rotate2Sphere(T, degree = 0) -} -\arguments{ - \item{T}{ -A matrix of test statistics. The rows correspond to protein/genes and the columns correspond to perturbations. The number of colmuns should be 2. -} - \item{degree}{ -The degree to be rotated. -} -} -\value{ -A transformed matrix with respect to the direction of interest. -} -\author{ -Pengyi Yang & Ellis Patrick -} - -\examples{ -# Load the example dataset. -data(PM) - -# Combine two perturbations into a single matrix. -# The matrix contains test statistics of each protein w.r.t each perturbation. -Tc <- cbind(Ins, Wmn) - -# Rotate the matrix by pi/2 in polar coordinates. -Tc.rotated <- rotate2Sphere(Tc, degree = pi/2) -} - - diff --git a/source/directPA_1.0/man/rotate3Sphere.Rd b/source/directPA_1.0/man/rotate3Sphere.Rd deleted file mode 100644 index 93b28dd..0000000 --- a/source/directPA_1.0/man/rotate3Sphere.Rd +++ /dev/null @@ -1,37 +0,0 @@ -\name{rotate3Sphere} -\alias{rotate3Sphere} -\title{ -Spherical Coordinates Rotation -} -\description{ -Rotate to the direction of interest in spherical coordinates by contrast (i.e 1, -1, -1). -} -\usage{ -rotate3Sphere(T, direction = c(1, 1, 1)) -} -\arguments{ - \item{T}{ -A matrix of test statistics. The rows correspond to protein/genes and the columns correspond to perturbations. The number of columns should be 3. -} - \item{direction}{ -The angle to be rotated in contrast format (see example below). -} -} -\value{ -A transformed matrix with respect to the direction of interest. -} -\author{ -Pengyi Yang & Ellis Patrick -} - -\examples{ -# Load the example data. -data(PM) - -# Combine three perturbations into a single matrix. -# The matrix contains test statistics of each protein w.r.t each perturbations. -Tc <- cbind(Ins, Wmn, MK) - -# Rotate the matrix by contrast 1, -1, -1 in spherical coordinates. -Tc.rotated <- rotate3Sphere(Tc, direction = c(1, -1, -1)) -} diff --git a/source/directPA_1.1/DESCRIPTION b/source/directPA_1.1/DESCRIPTION deleted file mode 100644 index 698fb4f..0000000 --- a/source/directPA_1.1/DESCRIPTION +++ /dev/null @@ -1,10 +0,0 @@ -Package: directPA -Type: Package -Title: Direction Pathway Analysis -Version: 1.1 -Date: 2015-03-13 -Author: Pengyi Yang & Ellis Patrick -Maintainer: Pengyi Yang -Description: A package for pathway or kinase analysis in experiments with multiple treatment designs. -License: GPL-3 -Depends: R (>= 2.10.0), rgl, calibrate diff --git a/source/directPA_1.1/R/directEnrichPlot2d.R b/source/directPA_1.1/R/directEnrichPlot2d.R deleted file mode 100644 index 44b660a..0000000 --- a/source/directPA_1.1/R/directEnrichPlot2d.R +++ /dev/null @@ -1,67 +0,0 @@ -#' Direction Enrichment Plot for Pathways and Kinases -#' -#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the -#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment -#' of pathways or kinases in direction specific manner. -#' -#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) -#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments -#' vs control statistics. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or -#' substrates belong to each pathway or kinase, respectively. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param ... parameters for controling the plot. -#' @return a list of coordinates for pathways or kinases -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' directEnrichPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3, xlim=c(-8, 3), ylim=c(-6, 3)) -#' -directEnrichPlot2d <- function(Tc, annotation, minSize=5, ...) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. filter the groups that are smaller than the minimun cutoff - DE = lapply(annotation, function(x){ - if(sum(rownames(Tc.zscores) %in% x) >= minSize) { - X <- Tc.zscores[rownames(Tc.zscores)%in%x,] - n = nrow(X) - Z1 = sum(X[,1])/sqrt(n) - Z2 = sum(X[,2])/sqrt(n) - list(Z1=Z1, Z2=Z2) - } - }) - - # step3. filter DE that has 0 element - DE <- DE[which(sapply(DE, length) != 0)] - Z1 <- unlist(sapply(DE, function(x){x[1]})) - Z2 <- unlist(sapply(DE, function(x){x[2]})) - - # visualization - plot(Z1, Z2, col="darkblue", pch=16, ...) - textxy(Z1,Z2, names(DE), col="black", cex=1) - abline(v=0, h=0, col="gold", lty=2) - abline(a=0, b=1, col="darkgreen", lty=2) - abline(a=0, b=-1, col="darkgreen", lty=2) - - r <- ceiling(max(sqrt(Z1^2 + Z2^2))) - for(i in seq(0, r, r/5)) { - theta = seq(-3.14,3.14,0.05) - lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") - } - - points(Z1, Z2, col="darkblue", pch=16, cex=2) - - ## return the results - result <- list() - result$Z1 <- Z1 - result$Z2 <- Z2 - return(result) -} diff --git a/source/directPA_1.1/R/directExplorer2d.R b/source/directPA_1.1/R/directExplorer2d.R deleted file mode 100644 index 41135d9..0000000 --- a/source/directPA_1.1/R/directExplorer2d.R +++ /dev/null @@ -1,92 +0,0 @@ -#' Batch Direction Analysis in 2-dimentional space -#' -#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) -#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns -#' are treatments vs control statistics. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to -#' genes or substrates belong to each pathway or kinase, respectively. -#' @param gene.method the method to be used for integrating statistics across treatments for each gene -#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is OSP. -#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation -#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is Stouffer. -#' @param top the number of entries to be highlighted in the plot. -#' @param ... parameters for controlling the plot. -#' @return The the list of enrichment analysis in tables. -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' # test enrichment on 8 directions in polar coordinate system. -#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) -#' -#' # the direction are denoted as follow for the two treatments vs control: -#' # ++: up-regulated in both treatments -#' # +*: up-regulated in the first treatment and unchanged in the second treatment -#' # +-: up-regulated in the first treatment and down-regulated in the second treatment -#' # *-: unchanged in the first treatment and down-regulated in the second treatment -#' # --: down-regulated in both treatments -#' # -*: down-regulated in the first treatment and unchanged in the second treatment -#' # -+: down-regulated in the first treatment and up-regulated in the second treatment -#' # *+: unchanged in the first treatment and up-regulated in the second treatment -#' -#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both -#' # treatments (i.e. "--") and displa the top-10 entries -#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] -#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] -#' -directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { - - directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') - gene.tab <- matrix(NA, nrow(Tc), 8) - colnames(gene.tab) <- directionCode - rownames(gene.tab) <- rownames(Tc) - - path.tab <- NULL - if (!is.null(annotation)) { - path.tab <- matrix(NA, length(annotation), 9) - colnames(path.tab) <- c("size", directionCode) - rownames(path.tab) <- names(annotation) - } - - for(i in 0:7){ - # gene or phosphorylation site level - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - gene.tab[,(i+1)] <- gene.pvalues - - # pathway or kinase level - if (!is.null(annotation)) { - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - if (i == 0) { - path.tab[,1] <- unlist(gst[,"size"]) - } - path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) - } - } - - # highlight candidates - plot(Tc, col="gray", pch=16, ...) - abline(h=0, v=0, col="gold", lty=2) - abline(a=0, b=1, lty = 2, col="darkgreen") - color <- rainbow(8) - for (i in 1:8) { - ids <- names(sort(gene.tab[,i])[1:top]) - points(Tc[ids,], col=color[i], pch=16) - textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) - } - - results <- list() - results$gene.tab <- gene.tab - results$path.tab <- path.tab - return(results) -} - diff --git a/source/directPA_1.1/R/directPA.R b/source/directPA_1.1/R/directPA.R deleted file mode 100644 index b365e57..0000000 --- a/source/directPA_1.1/R/directPA.R +++ /dev/null @@ -1,128 +0,0 @@ -#' Direction Analysis for Pathways and Kinases -#' -#' The main function of direction Analysis. This function takes in a matrix of test statistics with -#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and -#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for -#' enrichment of pathways or kinases on the specified direction. -#' -#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE) -#' -#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are -#' treatments vs control statistics. -#' @param direction the direction to be tested for enrichment. Either specified as a degree for -#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to -#' genes or substrates belong to each pathway or kinase, respectively. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param gene.method the method to be used for integrating statistics across treatments for each gene -#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is OSP. -#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation -#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is Stouffer. -#' @param visualize whether to visualize the -#' @return a list of enrichment for pathways or kinases -#' @export -#' @examples -#' -#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree -#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" -#' # (180 degree to original direction) -#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) -#' kst1[order(unlist(kst1[,1])),][1:20,] -#' -#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" -#' # (135 degree to the original direction) -#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) -#' kst2[order(unlist(kst2[,1])),][1:20,] -#' -#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" -#' # (225 degree to the original direction) -#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) -#' kst3[order(unlist(kst3[,1])),][1:20,] -#' -#' -#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) -#' # load the proteomics dataset -#' data(PM) -#' -#' # load pathway annotations -#' data(Pathways) -#' -#' # display reactome pathways. Could be replaced by any other pathway databases -#' Pathways.reactome[1:5] -#' -#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast -#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) -#' # on the original direction. -#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) -#' gst1[order(unlist(gst1[,1])),][1:20,] -#' -#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) -#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. -#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) -#' gst2[order(unlist(gst2[,1])),][1:20,] -#' -#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) -#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. -#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) -#' gst3[order(unlist(gst3[,1])),][1:20,] -#' -directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE){ - ## spherical coordinates for three-dimensional rotation - if (length(direction) == 3) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. rotate z-scores - Tc.rotated <- rotate3d(Tc.zscores, direction) - - # step 3. integrate statistics across treatments - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - - if (visualize == TRUE) { - HC = rainbow(length(gene.pvalues)*1.2) - plot3d(Tc, col=HC[rank(gene.pvalues)], size=5) - abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) - abclines3d(x=0, a=direction, col="pink", lwd=5) - } - - # step 4. integrate statistics for pathways or kinases - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - return(gst) - } - - ## polar coordinates for two-dimensional rotation - if (length(direction) == 1) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. rotate z-scores - Tc.rotated <- rotate2d(Tc.zscores, direction) - - # step 3. integrate statistics across treatments - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - - if (visualize == TRUE) { - HC = rainbow(length(gene.pvalues)*1.2) - plot(Tc, col=HC[rank(gene.pvalues)], pch=16) - abline(v = 0,h = 0, lty=2, col="gold") - } - - # step 4. integrate statistics for pathways or kinases - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - return(gst) - } -} diff --git a/source/directPA_1.1/R/geneStats.R b/source/directPA_1.1/R/geneStats.R deleted file mode 100644 index 2d296dc..0000000 --- a/source/directPA_1.1/R/geneStats.R +++ /dev/null @@ -1,43 +0,0 @@ -#' Gene Level Statistics -#' -#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, -#' and calculates a combined statistics accross multiple treatments. -#' -#' @usage geneStats(T, method="OSP") -#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control -#' comparison. -#' @param method the p-value integration method for combining accross multiple treatments. Available methods -#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. -#' @return a p-value after integration across treatments. -#' @export -#' -#' @examples -#' # load the example data -#' data(PM) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -#' # combine rotated statistics across treatments -#' gene.pvalues <- apply(PM.rotated, 1, geneStats) -#' -geneStats <- function(T, method="OSP") { - pvalue <- 0 - - if (method == "Stouffer") { - pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) - } else if (method == "OSP") { - p <- pnorm(T, lower.tail = TRUE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) - } else if (method == "Fisher") { - p <- pnorm(T, lower.tail = FALSE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) - } else if (method == "maxP") { - pvalue <- pnorm(max(T), lower.tail = FALSE) - } - - return (pvalue) -} diff --git a/source/directPA_1.1/R/pathwayStats.R b/source/directPA_1.1/R/pathwayStats.R deleted file mode 100644 index 0b6f177..0000000 --- a/source/directPA_1.1/R/pathwayStats.R +++ /dev/null @@ -1,63 +0,0 @@ -#' Pathway Level Statistics -#' -#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, -#' and calculates a combined statistics for those that belong to the same pathway or kinase. -#' -#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") -#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. -#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or -#' phosphorylation site that belong to the same pathway or kinase. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param method the p-value integration method for combining accross multiple treatments. Available methods -#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. -#' @return a doublet corresponding to the enrichment after integration across all genes or substrates -#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway -#' or kinase. -#' @export -#' -#' @examples -#' # load the example data -#' data(PM) -#' -#' # load pathway annotations -#' data(Pathways) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -#' # combine rotated statistics across treatments -#' gene.pvalues <- apply(PM.rotated, 1, geneStats) -#' -#' # compute statistics for all reactome pathways -#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) -#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) -#' -pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ - pvalue <- 0 - Z <- T[names(T) %in% PGs] - - if (length(Z) >= minSize) { - if (method == "Stouffer") { - pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) - } else if (method == "OSP") { - p <- pnorm(Z, lower.tail = TRUE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) - } else if (method == "Fisher") { - p <- pnorm(Z, lower.tail = FALSE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) - } else if (method == "maxP") { - pvalue <- pnorm(max(Z), lower.tail = FALSE) - } - } else { - pvalue <- NA - } - - result <- list() - result$pvalue <- pvalue - result$size <- length(Z) - return (result) -} diff --git a/source/directPA_1.1/R/rotate2d.R b/source/directPA_1.1/R/rotate2d.R deleted file mode 100644 index c6012ca..0000000 --- a/source/directPA_1.1/R/rotate2d.R +++ /dev/null @@ -1,26 +0,0 @@ -#' Polar Coordinates Rotation -#' -#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -#' @usage rotate2d(T, degree = 0) -#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation -#' sites and the columns are treatments vs control statistics. -#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. -#' @return A rotated matrix with respect to the direction of interest. -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # convert statistics into z-scores -#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) -#' -#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). -#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) -#' -rotate2d = function(T, degree = 0){ - theta = atan2(T[,2], T[,1]) + degree - r = sqrt(T[,1]^2 + T[,2]^2) - x2 = r*cos(theta) - y2 = r*sin(theta) - return(cbind(x2, y2)) -} diff --git a/source/directPA_1.1/R/rotate3d.R b/source/directPA_1.1/R/rotate3d.R deleted file mode 100644 index 10f8a79..0000000 --- a/source/directPA_1.1/R/rotate3d.R +++ /dev/null @@ -1,44 +0,0 @@ -#' Spherical Coordinates Rotation -#' -#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). -#' @usage rotate3d(T, contrast) -#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation -#' sites and the columns are treatments vs control statistics. -#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). -#' @return A rotated matrix with respect to the direction of interest. -#' @export -#' @examples -#' # load the example data -#' data(PM) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -rotate3d = function(T, contrast = c(1,1,1)){ - - a = contrast - b= c(1,1,1) # the original direction in contrast form is c(1,1,1) - - if(sum(a == b)==3)return(T) - if(sum(a == -b)==3)return(-T) - - a = a/sqrt(sum(a^2)) - b = b/sqrt(sum(b^2)) - u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) - u = u/sqrt(sum(u^2)) - R = matrix(0,3,3) - the = acos(sum(a*b)) - R[1,1] = cos(the) + u[1]^2*(1-cos(the)) - R[2,2] = cos(the) + u[2]^2*(1-cos(the)) - R[3,3] = cos(the) + u[3]^2*(1-cos(the)) - 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zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick -} -\references{ -Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, -David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. -} diff --git a/source/directPA_1.1/man/directPA.Rd b/source/directPA_1.1/man/directPA.Rd deleted file mode 100644 index 6fba82e..0000000 --- a/source/directPA_1.1/man/directPA.Rd +++ /dev/null @@ -1,92 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/directPA.R -\name{directPA} -\alias{directPA} -\title{Direction Analysis for Pathways and Kinases} -\usage{ -directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE) -} -\arguments{ -\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are -treatments vs control statistics.} - -\item{direction}{the direction to be tested for enrichment. Either specified as a degree for -two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} - -\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to -genes or substrates belong to each pathway or kinase, respectively.} - -\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups -that are smaller than the minSize will be removed from the analysis.} - -\item{gene.method}{the method to be used for integrating statistics across treatments for each gene -or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -Default method is OSP.} - -\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation -sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -Default method is Stouffer.} - -\item{visualize}{whether to visualize the} -} -\value{ -a list of enrichment for pathways or kinases -} -\description{ -The main function of direction Analysis. This function takes in a matrix of test statistics with -two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and -the annotation list such as pathway annotation or kinase-substrate annotation, and test for -enrichment of pathways or kinases on the specified direction. -} -\examples{ -## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) -# load the phosphoproteomics dataset -data(HEK) - -# load the kinase-substrate annoations -data(PhosphoSite) - -# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree -# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" -# (180 degree to original direction) -kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) -kst1[order(unlist(kst1[,1])),][1:20,] - -# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" -# (135 degree to the original direction) -kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) -kst2[order(unlist(kst2[,1])),][1:20,] - -# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" -# (225 degree to the original direction) -kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) -kst3[order(unlist(kst3[,1])),][1:20,] - - -## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) -# load the proteomics dataset -data(PM) - -# load pathway annotations -data(Pathways) - -# display reactome pathways. Could be replaced by any other pathway databases -Pathways.reactome[1:5] - -# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast -# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) -# on the original direction. -gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) -gst1[order(unlist(gst1[,1])),][1:20,] - -# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) -# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. -gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) -gst2[order(unlist(gst2[,1])),][1:20,] - -# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) -# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. -gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) -gst3[order(unlist(gst3[,1])),][1:20,] -} - diff --git a/source/directPA_1.1/man/geneStats.Rd b/source/directPA_1.1/man/geneStats.Rd deleted file mode 100644 index 455e3ef..0000000 --- a/source/directPA_1.1/man/geneStats.Rd +++ /dev/null @@ -1,36 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/geneStats.R -\name{geneStats} -\alias{geneStats} -\title{Gene Level Statistics} -\usage{ -geneStats(T, method="OSP") -} -\arguments{ -\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control -comparison.} - -\item{method}{the p-value integration method for combining accross multiple treatments. Available methods -are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} -} -\value{ -a p-value after integration across treatments. -} -\description{ -Takes a vector of statistics with each element corresponds to a treatment vs control comparison, -and calculates a combined statistics accross multiple treatments. -} -\examples{ -# load the example data -data(PM) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) - -# combine rotated statistics across treatments -gene.pvalues <- apply(PM.rotated, 1, geneStats) -} - diff --git a/source/directPA_1.1/man/pathwayStats.Rd b/source/directPA_1.1/man/pathwayStats.Rd deleted file mode 100644 index fec4ea5..0000000 --- a/source/directPA_1.1/man/pathwayStats.Rd +++ /dev/null @@ -1,50 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/pathwayStats.R -\name{pathwayStats} -\alias{pathwayStats} -\title{Pathway Level Statistics} -\usage{ -pathwayStats(PGs, T, minSize=5, method="Stouffer") -} -\arguments{ -\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} - -\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or -phosphorylation site that belong to the same pathway or kinase.} - -\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups -that are smaller than the minSize will be removed from the analysis.} - -\item{method}{the p-value integration method for combining accross multiple treatments. Available methods -are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} -} -\value{ -a doublet corresponding to the enrichment after integration across all genes or substrates -that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway -or kinase. -} -\description{ -Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, -and calculates a combined statistics for those that belong to the same pathway or kinase. -} -\examples{ -# load the example data -data(PM) - -# load pathway annotations -data(Pathways) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) - -# combine rotated statistics across treatments -gene.pvalues <- apply(PM.rotated, 1, geneStats) - -# compute statistics for all reactome pathways -gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) -gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) -} - diff --git a/source/directPA_1.1/man/rotate2d.Rd b/source/directPA_1.1/man/rotate2d.Rd deleted file mode 100644 index 940baa2..0000000 --- a/source/directPA_1.1/man/rotate2d.Rd +++ /dev/null @@ -1,31 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/rotate2d.R -\name{rotate2d} -\alias{rotate2d} -\title{Polar Coordinates Rotation} -\usage{ -rotate2d(T, degree = 0) -} -\arguments{ -\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation -sites and the columns are treatments vs control statistics.} - -\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} -} -\value{ -A rotated matrix with respect to the direction of interest. -} -\description{ -Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -} -\examples{ -# load the phosphoproteomics dataset -data(HEK) - -# convert statistics into z-scores -HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) - -# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). -HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) -} - diff --git a/source/directPA_1.1/man/rotate3d.Rd b/source/directPA_1.1/man/rotate3d.Rd deleted file mode 100644 index 8367f4c..0000000 --- a/source/directPA_1.1/man/rotate3d.Rd +++ /dev/null @@ -1,31 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/rotate3d.R -\name{rotate3d} -\alias{rotate3d} -\title{Spherical Coordinates Rotation} -\usage{ -rotate3d(T, contrast) -} -\arguments{ -\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation -sites and the columns are treatments vs control statistics.} - -\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} -} -\value{ -A rotated matrix with respect to the direction of interest. -} -\description{ -Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). -} -\examples{ -# load the example data -data(PM) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -} - diff --git a/source/directPA_1.2/.Rbuildignore b/source/directPA_1.2/.Rbuildignore deleted file mode 100644 index 91114bf..0000000 --- a/source/directPA_1.2/.Rbuildignore +++ /dev/null @@ -1,2 +0,0 @@ -^.*\.Rproj$ -^\.Rproj\.user$ diff --git a/source/directPA_1.2/.gitignore b/source/directPA_1.2/.gitignore deleted file mode 100644 index 807ea25..0000000 --- a/source/directPA_1.2/.gitignore +++ /dev/null @@ -1,3 +0,0 @@ -.Rproj.user -.Rhistory -.RData diff --git a/source/directPA_1.2/DESCRIPTION b/source/directPA_1.2/DESCRIPTION deleted file mode 100644 index c66409c..0000000 --- a/source/directPA_1.2/DESCRIPTION +++ /dev/null @@ -1,14 +0,0 @@ -Package: directPA -Type: Package -Title: Direction Analysis for Pathways and Kinases -Version: 1.2 -Date: 2015-06-18 -Author: Pengyi Yang & Ellis Patrick -Maintainer: Pengyi Yang -Description: Direction analysis is a set of tools designed to integrate multiple experimental treatments for analyzing pathways and kinases from microarray, RNA-seq, proteomics, or phosphoproteomics data. -License: GPL-3 -Depends: R (>= 2.10.0), rgl, calibrate -Packaged: 2015-06-18 13:08:32 UTC; Pengyi -NeedsCompilation: no -Repository: CRAN -Date/Publication: 2015-06-18 18:00:22 diff --git a/source/directPA_1.2/MD5 b/source/directPA_1.2/MD5 deleted file mode 100644 index 3c8c935..0000000 --- a/source/directPA_1.2/MD5 +++ /dev/null @@ -1,30 +0,0 @@ -e05721202d78707adb25b5dadb9a905c *DESCRIPTION -32f7448fafd8f272e98f8f3053cc3f51 *NAMESPACE -61911534dedf36247467a549fbf57ec1 *R/directEnrichPlot2d.R -76cad031899e402b3ab2c8ee3bad588f *R/directExplorer2d.R -c193001c14dbc241fd64be4f11202254 *R/directPA.R -016bae5eb003dc005c4dd37ccc50f58e *R/geneStats.R -c94800bbd93527de53b09fc6bbce19a3 *R/pathwayStats.R -0fb8763a7e2602a1004522f451bbea85 *R/rotate2d.R -989f12754c30a855e238dd4a160a9806 *R/rotate3d.R -83648a7a500619337f5f19913ad14f71 *data/HEK.rda -079cc541e6cfd73ded8ab5b20a8687e6 *data/PM.rda -d8b015f9de0c68ca738fa61b8604add3 *data/Pathways.rda -a4829f119c5d601a1663ff68b6f033c9 *data/PhosphoELM.rda -48bf3aaf03017c2519d62984ea9587a2 *data/PhosphoSite.rda -a8abd01dded8fe74481dfc5d6ee6284a *man/HEK.Rd -c02262b58e37b1fefd695441707b8809 *man/PM.Rd -19d4e0276d10cd41f6d577ed5245ea12 *man/Pathways.KEGG.Rd -ffd89f58f9af47d0ff556011a5c3521c *man/Pathways.reactome.Rd -50f5b7321b62385a4af7e5c1ee1ee901 *man/PhosphoELM.human.Rd -8e9a9b800cbdb9e5e9764143c54ef6fb *man/PhosphoELM.mouse.Rd -9224f661163899dbf13b3b3a8775a167 *man/PhosphoSite.human.Rd -02df716ef54fb50fdecd8648c6883235 *man/PhosphoSite.mouse.Rd -3d1ba4de7361b2947991bf8b6f1de90b *man/directEnrichPlot2d.Rd -29b9f292ce686fa8368ab99a2a527a74 *man/directExplorer2d.Rd -8809fdbeedb03dd0c8672be14ef11383 *man/directPA-package.Rd -02069d7456d6905cc361baab683de7f8 *man/directPA.Rd -fcc77f5e75cdbce83340ec66f2777f47 *man/geneStats.Rd -aefc1a7e9de81ead1d8f8d1911002812 *man/pathwayStats.Rd -3e13753f1c19b7fb239f048db2376b20 *man/rotate2d.Rd -574ba8d8e806479f12b078b52b6de344 *man/rotate3d.Rd diff --git a/source/directPA_1.2/NAMESPACE b/source/directPA_1.2/NAMESPACE deleted file mode 100644 index 4cdaa62..0000000 --- a/source/directPA_1.2/NAMESPACE +++ /dev/null @@ -1,11 +0,0 @@ -# Generated by roxygen2 (4.1.0): do not edit by hand - -export(perturbPlot2d) -export(directExplorer2d) -export(directPA) -export(geneStats) -export(pathwayStats) -export(rotate2d) -export(rotate3d) -import(rgl) -import(calibrate) diff --git a/source/directPA_1.2/R/directExplorer2d.R b/source/directPA_1.2/R/directExplorer2d.R deleted file mode 100644 index 0d8e08b..0000000 --- a/source/directPA_1.2/R/directExplorer2d.R +++ /dev/null @@ -1,93 +0,0 @@ -#' Batch Direction Analysis in 2-dimentional space -#' -#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -#' @usage directExplorer2d(Tc, annotation=NULL, gene.method="OSP", -#' path.method="Stouffer", top=10, ...) -#' @param Tc a numeric matrix with 2 columns. The rows are genes or phosphorylation sites and the columns -#' are treatments vs control statistics. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to -#' genes or substrates belong to each pathway or kinase, respectively. -#' @param gene.method the method to be used for integrating statistics across treatments for each gene -#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is OSP. -#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation -#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is Stouffer. -#' @param top the number of entries to be highlighted in the plot. -#' @param ... parameters for controlling the plot. -#' @return The the list of enrichment analysis in tables. -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' # test enrichment on 8 directions in polar coordinate system. -#' bda <- directExplorer2d(Tc=HEK, annotation=PhosphoSite.mouse) -#' -#' # the direction are denoted as follow for the two treatments vs control: -#' # ++: up-regulated in both treatments -#' # +*: up-regulated in the first treatment and unchanged in the second treatment -#' # +-: up-regulated in the first treatment and down-regulated in the second treatment -#' # *-: unchanged in the first treatment and down-regulated in the second treatment -#' # --: down-regulated in both treatments -#' # -*: down-regulated in the first treatment and unchanged in the second treatment -#' # -+: down-regulated in the first treatment and up-regulated in the second treatment -#' # *+: unchanged in the first treatment and up-regulated in the second treatment -#' -#' # sort the most enriched phosphorylation sites and kinases on down-regulaiton from both -#' # treatments (i.e. "--") and displa the top-10 entries -#' bda$gene.tab[order(bda$gene.tab[,"--"]),][1:10,] -#' bda$path.tab[order(bda$path.tab[,"--"]),][1:10,] -#' -directExplorer2d <- function(Tc, annotation=NULL, gene.method="OSP", path.method="Stouffer", top=10, ...) { - - directionCode <- c('++','+*','+-','*-','--','-*','-+','*+') - gene.tab <- matrix(NA, nrow(Tc), 8) - colnames(gene.tab) <- directionCode - rownames(gene.tab) <- rownames(Tc) - - path.tab <- NULL - if (!is.null(annotation)) { - path.tab <- matrix(NA, length(annotation), 9) - colnames(path.tab) <- c("size", directionCode) - rownames(path.tab) <- names(annotation) - } - - for(i in 0:7){ - # gene or phosphorylation site level - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - Tc.rotated <- rotate2d(Tc.zscores, pi/4*i) - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - gene.tab[,(i+1)] <- gene.pvalues - - # pathway or kinase level - if (!is.null(annotation)) { - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - if (i == 0) { - path.tab[,1] <- unlist(gst[,"size"]) - } - path.tab[,(i+2)] <- unlist(gst[,"pvalue"]) - } - } - - # highlight candidates - plot(Tc, col="gray", pch=16, ...) - abline(h=0, v=0, col="gold", lty=2) - abline(a=0, b=1, lty = 2, col="darkgreen") - color <- rainbow(8) - for (i in 1:8) { - ids <- names(sort(gene.tab[,i])[1:top]) - points(Tc[ids,], col=color[i], pch=16) - textxy(Tc[ids,1], Tc[ids,2], ids, col = color[i]) - } - - results <- list() - results$gene.tab <- gene.tab - results$path.tab <- path.tab - return(results) -} - diff --git a/source/directPA_1.2/R/directPA.R b/source/directPA_1.2/R/directPA.R deleted file mode 100644 index 0e2b4e4..0000000 --- a/source/directPA_1.2/R/directPA.R +++ /dev/null @@ -1,139 +0,0 @@ -#' Direction Analysis for Pathways and Kinases -#' -#' The main function of direction Analysis. This function takes in a matrix of test statistics with -#' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and -#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for -#' enrichment of pathways or kinases on the specified direction. -#' -#' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", -#' path.method="Stouffer", visualize=TRUE) -#' -#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are -#' treatments vs control statistics. -#' @param direction the direction to be tested for enrichment. Either specified as a degree for -#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to -#' genes or substrates belong to each pathway or kinase, respectively. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param gene.method the method to be used for integrating statistics across treatments for each gene -#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is OSP. -#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation -#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is Stouffer. -#' @param visualize whether to visualize the -#' @return a list of enrichment for pathways or kinases -#' @export -#' @examples -#' -#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree -#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" -#' # (180 degree to original direction) -#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) -#' kst1[order(unlist(kst1[,1])),][1:20,] -#' -#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" -#' # (135 degree to the original direction) -#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) -#' kst2[order(unlist(kst2[,1])),][1:20,] -#' -#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" -#' # (225 degree to the original direction) -#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) -#' kst3[order(unlist(kst3[,1])),][1:20,] -#' -#' -#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) -#' # load the proteomics dataset -#' data(PM) -#' -#' # load pathway annotations -#' data(Pathways) -#' -#' # display reactome pathways. Could be replaced by any other pathway databases -#' Pathways.reactome[1:5] -#' -#' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast -#' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) -#' # on the original direction. -#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) -#' gst1[order(unlist(gst1[,1])),][1:20,] -#' -#' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) -#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. -#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) -#' gst2[order(unlist(gst2[,1])),][1:20,] -#' -#' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) -#' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. -#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) -#' gst3[order(unlist(gst3[,1])),][1:20,] -#' -directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE, ...){ - ## spherical coordinates for three-dimensional rotation - if (length(direction) == 3) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. rotate z-scores - Tc.rotated <- rotate3d(Tc.zscores, direction) - - # step 3. integrate statistics across treatments - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - - if (visualize == TRUE) { - HC = rainbow(length(gene.pvalues)*1.2) - plot3d(Tc, col=HC[rank(gene.pvalues)], size=5, ...) - abclines3d(x=0, y=0, z=0, a=diag(3), col="black", lwd=3) - abclines3d(x=0, a=direction, col="pink", lwd=5) - } - - # step 4. integrate statistics for pathways or kinases - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - - result <- list() - result$gene.pvalues <- gene.pvalues - result$gst <- gst - return(result) - } - - ## polar coordinates for two-dimensional rotation - if (length(direction) == 1) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. rotate z-scores - Tc.rotated <- rotate2d(Tc.zscores, direction) - - # step 3. integrate statistics across treatments - gene.pvalues <- apply(Tc.rotated, 1, geneStats, gene.method) - - if (visualize == TRUE) { - HC = rainbow(length(gene.pvalues)*1.2) - plot(Tc, col=HC[rank(gene.pvalues)], pch=16, ...) - abline(v = 0,h = 0, lty=2, col="gold") - abline(a=0, b=1, col="darkgreen", lty=2) - abline(a=0, b=-1, col="darkgreen", lty=2) - } - - # step 4. integrate statistics for pathways or kinases - gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) - gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) - - result <- list() - result$gene.pvalues <- gene.pvalues - result$gst <- gst - return(result) - } -} diff --git a/source/directPA_1.2/R/geneStats.R b/source/directPA_1.2/R/geneStats.R deleted file mode 100644 index 2d296dc..0000000 --- a/source/directPA_1.2/R/geneStats.R +++ /dev/null @@ -1,43 +0,0 @@ -#' Gene Level Statistics -#' -#' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, -#' and calculates a combined statistics accross multiple treatments. -#' -#' @usage geneStats(T, method="OSP") -#' @param T a vector of statistics (z-scores converted) with each element correspond to a treatment vs control -#' comparison. -#' @param method the p-value integration method for combining accross multiple treatments. Available methods -#' are Stouffer, OSP, Fisher, and maxP. The default method is OSP. -#' @return a p-value after integration across treatments. -#' @export -#' -#' @examples -#' # load the example data -#' data(PM) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -#' # combine rotated statistics across treatments -#' gene.pvalues <- apply(PM.rotated, 1, geneStats) -#' -geneStats <- function(T, method="OSP") { - pvalue <- 0 - - if (method == "Stouffer") { - pvalue <- pnorm(sum(T), 0, sqrt(length(T)), lower.tail =FALSE) - } else if (method == "OSP") { - p <- pnorm(T, lower.tail = TRUE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) - } else if (method == "Fisher") { - p <- pnorm(T, lower.tail = FALSE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) - } else if (method == "maxP") { - pvalue <- pnorm(max(T), lower.tail = FALSE) - } - - return (pvalue) -} diff --git a/source/directPA_1.2/R/pathwayStats.R b/source/directPA_1.2/R/pathwayStats.R deleted file mode 100644 index 107581f..0000000 --- a/source/directPA_1.2/R/pathwayStats.R +++ /dev/null @@ -1,63 +0,0 @@ -#' Pathway Level Statistics -#' -#' Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, -#' and calculates a combined statistics for those that belong to the same pathway or kinase. -#' -#' @usage pathwayStats(PGs, T, minSize=5, method="Stouffer") -#' @param PGs an array of names indicating genes or substrates that belong to a given pathway or kinase. -#' @param T a vector of statistics (z-scores converted) with each element correspond to a gene or -#' phosphorylation site that belong to the same pathway or kinase. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param method the p-value integration method for combining accross multiple treatments. Available methods -#' are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer. -#' @return a doublet corresponding to the enrichment after integration across all genes or substrates -#' that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway -#' or kinase. -#' @export -#' -#' @examples -#' # load the example data -#' data(PM) -#' -#' # load pathway annotations -#' data(Pathways) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -#' # combine rotated statistics across treatments -#' gene.pvalues <- apply(PM.rotated, 1, geneStats) -#' -#' # compute statistics for all reactome pathways -#' gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) -#' gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) -#' -pathwayStats = function(PGs, T, minSize=5, method="Stouffer"){ - pvalue <- 0 - Z <- T[names(T) %in% PGs] - - if (length(Z) >= minSize) { - if (method == "Stouffer") { - pvalue <- pnorm(sum(Z), 0, sqrt(length(Z)), lower.tail = FALSE) - } else if (method == "OSP") { - p <- pnorm(Z, lower.tail = TRUE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = TRUE) - } else if (method == "Fisher") { - p <- pnorm(Z, lower.tail = FALSE) - pvalue <- pchisq(-2*sum(log(p)), 2*length(p), lower.tail = FALSE) - } else if (method == "maxP") { - pvalue <- pnorm(max(Z), lower.tail = FALSE) - } - } else { - pvalue <- NA - } - - result <- list() - result$pvalue <- pvalue - result$size <- length(Z) - return (result) -} diff --git a/source/directPA_1.2/R/perturbPlot2d.R b/source/directPA_1.2/R/perturbPlot2d.R deleted file mode 100644 index b46dcfb..0000000 --- a/source/directPA_1.2/R/perturbPlot2d.R +++ /dev/null @@ -1,67 +0,0 @@ -#' Perturbation Plot -#' -#' This function takes in a matrix of test statistics with two columns (2-dimensional space) and the -#' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment -#' of pathways or kinases in direction specific manner. -#' -#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) -#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments -#' vs control statistics. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or -#' substrates belong to each pathway or kinase, respectively. -#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups -#' that are smaller than the minSize will be removed from the analysis. -#' @param ... parameters for controling the plot. -#' @return a list of coordinates for pathways or kinases -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' perturbPlot2d(Tc=HEK, annotation=PhosphoSite.mouse, cex=3) -#' -perturbPlot2d <- function(Tc, annotation, minSize=5, ...) { - - # step 1. convert statistics into z-scores - Tc.zscores <- apply(Tc, 2, function(x){qnorm(rank(x)/(nrow(Tc)+1))}) - - # step 2. filter the groups that are smaller than the minimun cutoff - DE = lapply(annotation, function(x){ - if(sum(rownames(Tc.zscores) %in% x) >= minSize) { - X <- Tc.zscores[rownames(Tc.zscores)%in%x,] - n = nrow(X) - Z1 = sum(X[,1])/sqrt(n) - Z2 = sum(X[,2])/sqrt(n) - list(Z1=Z1, Z2=Z2) - } - }) - - # step3. filter DE that has 0 element - DE <- DE[which(sapply(DE, length) != 0)] - Z1 <- unlist(sapply(DE, function(x){x[1]})) - Z2 <- unlist(sapply(DE, function(x){x[2]})) - - # visualization - plot(Z1, Z2, col="darkblue", pch=16, xlab=colnames(Tc)[1], ylab=colnames(Tc)[2], ...) - textxy(Z1,Z2, names(DE), col="black", cex=1) - abline(v=0, h=0, col="gold", lty=2) - abline(a=0, b=1, col="darkgreen", lty=2) - abline(a=0, b=-1, col="darkgreen", lty=2) - - r <- ceiling(max(sqrt(Z1^2 + Z2^2))) - for(i in seq(0, r, r/5)) { - theta = seq(-3.14,3.14,0.05) - lines(i*cos(theta),i*sin(theta),col = 'gray', type="l") - } - - points(Z1, Z2, col="darkblue", pch=16, cex=2) - - ## return the results - result <- list() - result$Z1 <- Z1 - result$Z2 <- Z2 - return(result) -} diff --git a/source/directPA_1.2/R/rotate2d.R b/source/directPA_1.2/R/rotate2d.R deleted file mode 100644 index c6012ca..0000000 --- a/source/directPA_1.2/R/rotate2d.R +++ /dev/null @@ -1,26 +0,0 @@ -#' Polar Coordinates Rotation -#' -#' Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -#' @usage rotate2d(T, degree = 0) -#' @param T a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation -#' sites and the columns are treatments vs control statistics. -#' @param degree the direction to be tested for enrichment. Specified as degree from to the original direction. -#' @return A rotated matrix with respect to the direction of interest. -#' @export -#' @examples -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # convert statistics into z-scores -#' HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) -#' -#' # Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). -#' HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) -#' -rotate2d = function(T, degree = 0){ - theta = atan2(T[,2], T[,1]) + degree - r = sqrt(T[,1]^2 + T[,2]^2) - x2 = r*cos(theta) - y2 = r*sin(theta) - return(cbind(x2, y2)) -} diff --git a/source/directPA_1.2/R/rotate3d.R b/source/directPA_1.2/R/rotate3d.R deleted file mode 100644 index 10f8a79..0000000 --- a/source/directPA_1.2/R/rotate3d.R +++ /dev/null @@ -1,44 +0,0 @@ -#' Spherical Coordinates Rotation -#' -#' Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). -#' @usage rotate3d(T, contrast) -#' @param T a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation -#' sites and the columns are treatments vs control statistics. -#' @param contrast the direction to be tested for enrichment. Specified as contrast in a triplet (see example). -#' @return A rotated matrix with respect to the direction of interest. -#' @export -#' @examples -#' # load the example data -#' data(PM) -#' -#' # convert statistics into z-scores -#' PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) -#' -#' # Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -#' PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -#' -rotate3d = function(T, contrast = c(1,1,1)){ - - a = contrast - b= c(1,1,1) # the original direction in contrast form is c(1,1,1) - - if(sum(a == b)==3)return(T) - if(sum(a == -b)==3)return(-T) - - a = a/sqrt(sum(a^2)) - b = b/sqrt(sum(b^2)) - u = c((a[2]*b[3] - a[3]*b[2]) , (a[3]*b[1] - a[1]*b[3]) ,(a[1]*b[2] - a[2]*b[1]) ) - u = u/sqrt(sum(u^2)) - R = matrix(0,3,3) - the = acos(sum(a*b)) - R[1,1] = cos(the) + u[1]^2*(1-cos(the)) - R[2,2] = cos(the) + u[2]^2*(1-cos(the)) - R[3,3] = cos(the) + u[3]^2*(1-cos(the)) - 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zL80D?idv`n{h{jM%DBslXiOX7kg=a*-#ZID{S`)(-bM}+M0!(Ou~M2 zR~=WKI4MDJQ9XLeQ60yM*7&TsFcBuz&nA+#MBfD*|H&^y zr>Ftr|HnqFz%IZd7p-kJVq7m6hs{K8Dg+=^R>izt$LLxn1T zh@^;~SvrT$N^04HCUD=*W0t=v+z}}usOBWS^IJqi63kKZXE=Szv5 zXXF`M+VnX|D-u%7=J8d=op487T9sFu8=%!{h^LnIrbLM)w?ominYr7KsC_vbo=fN4 zHi#fl&v)Mpq5GB;wNZ;hjh8u2_4+Vn%~DMiipZ&b4{2N%s~nO&hTd}%ag5;XMT3cv zOyx)+Po4~B=4p>#uOF-CjObeL-gNlG0b6Y?x4D7Q;NJVATq7yp#Olv|Cp$32tbR5M*AOU>Iap!gYL)N_6Oce$8}DY>jE#0$egd8h()CXp7<#x7 zJ;MLWMCb3nl+QXu^j>yt@l4wlD-KN*`u zzNBXx(L7_lt87iX zTsA0PW-^WwN4R1ixCJC7cY_}*%;igRvvLh|K0&q-xO2&}3JAazLJ_OIWH~>ZRdI0M z!KVIs`>{Zs2moe1^&wVwt?jx)UYg;;;NNX!tIPB0d?wZ``LQDC;{dP!cBtGvx}Bce zrgOU$diYLs_7;&AXTd8YL(B*O5Vxr6#C2#o;zGnLP}-vGR|DaC{1M_K^kl1Kr+S^h zRqjDB{F-0%0}mIJK18nW(H3Oe47`uaAthUp6SN|`uI+( & Ellis Patrick -} -\references{ -Pengyi Yang, Ellis Patrick, Shi-Xiong Tan, Daniel J. Fazakerley, James Burchfield, Christopher Gribben, Matthew J. Prior, -David E. James, Yee Hwa Yang, Direction pathway analysis of large-scale proteomics data reveals novel features of the insulin action pathway, submitted. -} diff --git a/source/directPA_1.2/man/directPA.Rd b/source/directPA_1.2/man/directPA.Rd deleted file mode 100644 index 50ffbeb..0000000 --- a/source/directPA_1.2/man/directPA.Rd +++ /dev/null @@ -1,93 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/directPA.R -\name{directPA} -\alias{directPA} -\title{Direction Analysis for Pathways and Kinases} -\usage{ -directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", -path.method="Stouffer", visualize=TRUE) -} -\arguments{ -\item{Tc}{a numeric matrix. The columns are genes or phosphorylation sites and the columns are -treatments vs control statistics.} - -\item{direction}{the direction to be tested for enrichment. Either specified as a degree for -two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis.} - -\item{annotation}{a list with names correspond to pathways or kinases and elements correspond to -genes or substrates belong to each pathway or kinase, respectively.} - -\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups -that are smaller than the minSize will be removed from the analysis.} - -\item{gene.method}{the method to be used for integrating statistics across treatments for each gene -or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. -Default method is OSP.} - -\item{path.method}{the method to be used for integrating statistics of all genes or phosphorylation -sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -Default method is Stouffer.} - -\item{visualize}{whether to visualize the} -} -\value{ -a list of enrichment for pathways or kinases -} -\description{ -The main function of direction Analysis. This function takes in a matrix of test statistics with -two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and -the annotation list such as pathway annotation or kinase-substrate annotation, and test for -enrichment of pathways or kinases on the specified direction. -} -\examples{ -## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) -# load the phosphoproteomics dataset -data(HEK) - -# load the kinase-substrate annoations -data(PhosphoSite) - -# direction pathway analysis in 2-dimensional space. Implemented as rotating by degree -# (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" -# (180 degree to original direction) -kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) -kst1[order(unlist(kst1[,1])),][1:20,] - -# (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" -# (135 degree to the original direction) -kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) -kst2[order(unlist(kst2[,1])),][1:20,] - -# (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" -# (225 degree to the original direction) -kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) -kst3[order(unlist(kst3[,1])),][1:20,] - - -## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) -# load the proteomics dataset -data(PM) - -# load pathway annotations -data(Pathways) - -# display reactome pathways. Could be replaced by any other pathway databases -Pathways.reactome[1:5] - -# direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast -# (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) -# on the original direction. -gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) -gst1[order(unlist(gst1[,1])),][1:20,] - -# (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) -# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. -gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) -gst2[order(unlist(gst2[,1])),][1:20,] - -# (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) -# this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. -gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) -gst3[order(unlist(gst3[,1])),][1:20,] -} - diff --git a/source/directPA_1.2/man/geneStats.Rd b/source/directPA_1.2/man/geneStats.Rd deleted file mode 100644 index 455e3ef..0000000 --- a/source/directPA_1.2/man/geneStats.Rd +++ /dev/null @@ -1,36 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/geneStats.R -\name{geneStats} -\alias{geneStats} -\title{Gene Level Statistics} -\usage{ -geneStats(T, method="OSP") -} -\arguments{ -\item{T}{a vector of statistics (z-scores converted) with each element correspond to a treatment vs control -comparison.} - -\item{method}{the p-value integration method for combining accross multiple treatments. Available methods -are Stouffer, OSP, Fisher, and maxP. The default method is OSP.} -} -\value{ -a p-value after integration across treatments. -} -\description{ -Takes a vector of statistics with each element corresponds to a treatment vs control comparison, -and calculates a combined statistics accross multiple treatments. -} -\examples{ -# load the example data -data(PM) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) - -# combine rotated statistics across treatments -gene.pvalues <- apply(PM.rotated, 1, geneStats) -} - diff --git a/source/directPA_1.2/man/pathwayStats.Rd b/source/directPA_1.2/man/pathwayStats.Rd deleted file mode 100644 index fec4ea5..0000000 --- a/source/directPA_1.2/man/pathwayStats.Rd +++ /dev/null @@ -1,50 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/pathwayStats.R -\name{pathwayStats} -\alias{pathwayStats} -\title{Pathway Level Statistics} -\usage{ -pathwayStats(PGs, T, minSize=5, method="Stouffer") -} -\arguments{ -\item{PGs}{an array of names indicating genes or substrates that belong to a given pathway or kinase.} - -\item{T}{a vector of statistics (z-scores converted) with each element correspond to a gene or -phosphorylation site that belong to the same pathway or kinase.} - -\item{minSize}{the size of annotation groups to be considered for calculating enrichment. Groups -that are smaller than the minSize will be removed from the analysis.} - -\item{method}{the p-value integration method for combining accross multiple treatments. Available methods -are Stouffer, OSP, Fisher, and maxP. The default method is Stouffer.} -} -\value{ -a doublet corresponding to the enrichment after integration across all genes or substrates -that belong to the same pathway or kinase, and the size of the mapped genes or substrates to that pathway -or kinase. -} -\description{ -Takes a vector of statistics with each element corresponds to a gene or phosphorylation site, -and calculates a combined statistics for those that belong to the same pathway or kinase. -} -\examples{ -# load the example data -data(PM) - -# load pathway annotations -data(Pathways) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) - -# combine rotated statistics across treatments -gene.pvalues <- apply(PM.rotated, 1, geneStats) - -# compute statistics for all reactome pathways -gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) -gst <- t(sapply(Pathways.reactome, pathwayStats, gene.zscores)) -} - diff --git a/source/directPA_1.2/man/rotate2d.Rd b/source/directPA_1.2/man/rotate2d.Rd deleted file mode 100644 index 940baa2..0000000 --- a/source/directPA_1.2/man/rotate2d.Rd +++ /dev/null @@ -1,31 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/rotate2d.R -\name{rotate2d} -\alias{rotate2d} -\title{Polar Coordinates Rotation} -\usage{ -rotate2d(T, degree = 0) -} -\arguments{ -\item{T}{a numeric matrix (normally z-score converted) with 2 columns. The rows are genes or phosphorylation -sites and the columns are treatments vs control statistics.} - -\item{degree}{the direction to be tested for enrichment. Specified as degree from to the original direction.} -} -\value{ -A rotated matrix with respect to the direction of interest. -} -\description{ -Rotate to the direction of interest in polar coordinates by degree (e.g. pi/4). -} -\examples{ -# load the phosphoproteomics dataset -data(HEK) - -# convert statistics into z-scores -HEK.zscores <- apply(HEK, 2, function(x){qnorm(rank(x)/(nrow(HEK)+1))}) - -# Rotate the matrix by 1/2 pi (i.e. down-regulation, dow-regulation). -HEK.rotated <- rotate2d(HEK.zscores, degree = pi/2) -} - diff --git a/source/directPA_1.2/man/rotate3d.Rd b/source/directPA_1.2/man/rotate3d.Rd deleted file mode 100644 index 8367f4c..0000000 --- a/source/directPA_1.2/man/rotate3d.Rd +++ /dev/null @@ -1,31 +0,0 @@ -% Generated by roxygen2 (4.1.0): do not edit by hand -% Please edit documentation in R/rotate3d.R -\name{rotate3d} -\alias{rotate3d} -\title{Spherical Coordinates Rotation} -\usage{ -rotate3d(T, contrast) -} -\arguments{ -\item{T}{a numeric matrix (normally z-score converted) with 3 columns. The rows are genes or phosphorylation -sites and the columns are treatments vs control statistics.} - -\item{contrast}{the direction to be tested for enrichment. Specified as contrast in a triplet (see example).} -} -\value{ -A rotated matrix with respect to the direction of interest. -} -\description{ -Rotate to the direction of interest in spherical coordinates by contrasts (e.g. 1, -1, -1). -} -\examples{ -# load the example data -data(PM) - -# convert statistics into z-scores -PM.zscores <- apply(PM, 2, function(x){qnorm(rank(x)/(nrow(PM)+1))}) - -# Rotate the matrix by contrast 1, -1, -1 (i.e. up-regulation, down-regulation, dow-regulation). -PM.rotated <- rotate3d(PM.zscores, contrast = c(1, -1, -1)) -} - From cbeb26e1a2e86fd4f1f57e7e8d799aade508fb30 Mon Sep 17 00:00:00 2001 From: PengyiYang Date: Tue, 17 Nov 2015 16:04:24 +1100 Subject: [PATCH 06/12] remove --- source/directPA_1.1/NAMESPACE | 11 ----------- 1 file changed, 11 deletions(-) delete mode 100644 source/directPA_1.1/NAMESPACE diff --git a/source/directPA_1.1/NAMESPACE b/source/directPA_1.1/NAMESPACE deleted file mode 100644 index 7fe39f5..0000000 --- a/source/directPA_1.1/NAMESPACE +++ /dev/null @@ -1,11 +0,0 @@ -# Generated by roxygen2 (4.1.0): do not edit by hand - -export(directEnrichPlot2d) -export(directExplorer2d) -export(directPA) -export(geneStats) -export(pathwayStats) -export(rotate2d) -export(rotate3d) -import(rgl) -import(calibrate) From 5c3eba31a0a050288d278add4469800713b65213 Mon Sep 17 00:00:00 2001 From: PengyiYang Date: Tue, 22 Mar 2016 15:46:41 +1100 Subject: [PATCH 07/12] update package 1.3; 22 March 2016 --- DESCRIPTION | 19 +- LICENSE | 675 ------------------ NAMESPACE | 5 +- R/directPA.R | 73 +- R/geneStats.R | 2 +- R/kinasePA.R | 59 ++ R/perturbPlot2d.R | 2 +- README.md | 2 +- directPA_1.0.tar.gz | Bin 98092 -> 0 bytes directPA_1.0.zip | Bin 119002 -> 0 bytes man/directExplorer2d.Rd | 15 +- man/directPA.Rd | 82 +-- man/geneStats.Rd | 11 +- man/kinasePA.Rd | 69 ++ man/pathwayStats.Rd | 13 +- ...directEnrichPlot2d.Rd => perturbPlot2d.Rd} | 21 +- man/rotate2d.Rd | 3 +- man/rotate3d.Rd | 3 +- 18 files changed, 239 insertions(+), 815 deletions(-) delete mode 100644 LICENSE create mode 100644 R/kinasePA.R delete mode 100644 directPA_1.0.tar.gz delete mode 100644 directPA_1.0.zip create mode 100644 man/kinasePA.Rd rename man/{directEnrichPlot2d.Rd => perturbPlot2d.Rd} (64%) diff --git a/DESCRIPTION b/DESCRIPTION index c66409c..2424bd3 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -1,14 +1,21 @@ Package: directPA Type: Package Title: Direction Analysis for Pathways and Kinases -Version: 1.2 -Date: 2015-06-18 +Version: 1.3 +Date: 2016-03-22 Author: Pengyi Yang & Ellis Patrick Maintainer: Pengyi Yang -Description: Direction analysis is a set of tools designed to integrate multiple experimental treatments for analyzing pathways and kinases from microarray, RNA-seq, proteomics, or phosphoproteomics data. +Description: Direction analysis is a set of tools designed to identify + combinatorial effects of multiple treatments and/or perturbations on pathways + and kinases profiled by microarray, RNA-seq, proteomics, or phosphoproteomics + data. License: GPL-3 -Depends: R (>= 2.10.0), rgl, calibrate -Packaged: 2015-06-18 13:08:32 UTC; Pengyi +Depends: + R (>= 2.10.0), + rgl, + calibrate +Packaged: 2016-03-22 13:08:32 UTC; Pengyi NeedsCompilation: no Repository: CRAN -Date/Publication: 2015-06-18 18:00:22 +Date/Publication: 2016-03-22 18:00:22 +RoxygenNote: 5.0.1 diff --git a/LICENSE b/LICENSE deleted file mode 100644 index 6b156fe..0000000 --- a/LICENSE +++ /dev/null @@ -1,675 +0,0 @@ -GNU GENERAL PUBLIC LICENSE - Version 3, 29 June 2007 - - Copyright (C) 2007 Free Software Foundation, Inc. - Everyone is permitted to copy and distribute verbatim copies - of this license document, but changing it is not allowed. - - Preamble - - The GNU General Public License is a free, copyleft license for -software and other kinds of works. - - The licenses for most software and other practical works are designed -to take away your freedom to share and change the works. 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Of course, your program's commands -might be different; for a GUI interface, you would use an "about box". - - You should also get your employer (if you work as a programmer) or school, -if any, to sign a "copyright disclaimer" for the program, if necessary. -For more information on this, and how to apply and follow the GNU GPL, see -. - - The GNU General Public License does not permit incorporating your program -into proprietary programs. If your program is a subroutine library, you -may consider it more useful to permit linking proprietary applications with -the library. If this is what you want to do, use the GNU Lesser General -Public License instead of this License. But first, please read -. - diff --git a/NAMESPACE b/NAMESPACE index 4cdaa62..75bbc6c 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -1,10 +1,11 @@ -# Generated by roxygen2 (4.1.0): do not edit by hand +# Generated by roxygen2: do not edit by hand -export(perturbPlot2d) export(directExplorer2d) export(directPA) export(geneStats) +export(kinasePA) export(pathwayStats) +export(perturbPlot2d) export(rotate2d) export(rotate3d) import(rgl) diff --git a/R/directPA.R b/R/directPA.R index 0e2b4e4..512de3c 100644 --- a/R/directPA.R +++ b/R/directPA.R @@ -1,57 +1,32 @@ -#' Direction Analysis for Pathways and Kinases +#' Direction Analysis for Pathways #' #' The main function of direction Analysis. This function takes in a matrix of test statistics with #' two (2-dimensional space) or three (3-dimensional space) columns, the direction of interests, and -#' the annotation list such as pathway annotation or kinase-substrate annotation, and test for -#' enrichment of pathways or kinases on the specified direction. +#' the annotation list such as pathway annotation, and test for enrichment of pathways on the specified +#' direction. #' #' @usage directPA(Tc, direction, annotation, minSize=5, gene.method="OSP", -#' path.method="Stouffer", visualize=TRUE) +#' path.method="Stouffer", visualize=TRUE, ...) #' -#' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are -#' treatments vs control statistics. +#' @param Tc a numeric matrix. The columns are genes and the columns are treatments vs control statistics. #' @param direction the direction to be tested for enrichment. Either specified as a degree for #' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. -#' @param annotation a list with names correspond to pathways or kinases and elements correspond to -#' genes or substrates belong to each pathway or kinase, respectively. +#' @param annotation a list with names correspond to pathways and elements correspond to genes belong to +#' each pathway, respectively. #' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups #' that are smaller than the minSize will be removed from the analysis. -#' @param gene.method the method to be used for integrating statistics across treatments for each gene -#' or phosphorylation site. Available methods are Stouffer, OSP, Fisher, and maxP. +#' @param gene.method the method to be used for integrating statistics across treatments for each gene. +#' Available methods are Stouffer, OSP, Fisher, and maxP. #' Default method is OSP. -#' @param path.method the method to be used for integrating statistics of all genes or phosphorylation -#' sites that belongs to a pathway or kinase. Available methods are Stouffer, OSP, Fisher, and maxP. -#' Default method is Stouffer. -#' @param visualize whether to visualize the -#' @return a list of enrichment for pathways or kinases +#' @param path.method the method to be used for integrating statistics of all genes that belongs to a +#' pathway. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the plot. +#' @param ... other visualization parameters to pass on. +#' @return a list that contains directional p-values for each gene and directional enrichment for each pathway. #' @export #' @examples #' -#' ## I. for kinase anlaysis on a phosphoproteomics dataset (Humphrey et al. Cell Metab., 2013) -#' # load the phosphoproteomics dataset -#' data(HEK) -#' -#' # load the kinase-substrate annoations -#' data(PhosphoSite) -#' -#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree -#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" -#' # (180 degree to original direction) -#' kst1 <- directPA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) -#' kst1[order(unlist(kst1[,1])),][1:20,] -#' -#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" -#' # (135 degree to the original direction) -#' kst2 <- directPA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) -#' kst2[order(unlist(kst2[,1])),][1:20,] -#' -#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" -#' # (225 degree to the original direction) -#' kst3 <- directPA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) -#' kst3[order(unlist(kst3[,1])),][1:20,] -#' -#' -#' ## II. for pathway analysis on a proteomics dataset (Yang et al. Bioinformatics, 2014) #' # load the proteomics dataset #' data(PM) #' @@ -64,18 +39,20 @@ #' # direction pathway analysis in 3-dimensional space. Implemnted as rotating by contrast #' # (1) test combined effect of all 3 treatments (stimulation and inhibitions) vs control (basal) #' # on the original direction. -#' gst1 <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) -#' gst1[order(unlist(gst1[,1])),][1:20,] +#' dPA <- directPA(Tc=PM, direction=c(1,1,1), annotation=Pathways.reactome) +#' dPA$gst[order(unlist(dPA$gst[,1])),][1:20,] +#' # rank substrates on the direciton of interest +#' sort(dPA$gene.pvalues)[1:20] #' #' # (2) test combined effect of all 3 treatments vs controls on direction c(1,-1, 0) #' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 45 degree. -#' gst2 <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) -#' gst2[order(unlist(gst2[,1])),][1:20,] +#' dPA <- directPA(Tc=PM, direction=c(1,-1,0), annotation=Pathways.reactome) +#' dPA$gst[order(unlist(dPA$gst[,1])),][1:20,] #' #' # (3) test combined effect of all 3 perturbations vs controls on direction c(1,-1, 1) #' # this rotates Ins by 0 degree, Wmn by 90 degree, and MK by 0 degree. -#' gst3 <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) -#' gst3[order(unlist(gst3[,1])),][1:20,] +#' dPA <- directPA(Tc=PM, direction=c(1,-1,1), annotation=Pathways.reactome) +#' dPA$gst[order(unlist(dPA$gst[,1])),][1:20,] #' directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", path.method="Stouffer", visualize=TRUE, ...){ ## spherical coordinates for three-dimensional rotation @@ -97,7 +74,7 @@ directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", pa abclines3d(x=0, a=direction, col="pink", lwd=5) } - # step 4. integrate statistics for pathways or kinases + # step 4. integrate statistics for pathways gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) @@ -127,7 +104,7 @@ directPA <- function(Tc, direction, annotation, minSize=5, gene.method="OSP", pa abline(a=0, b=-1, col="darkgreen", lty=2) } - # step 4. integrate statistics for pathways or kinases + # step 4. integrate statistics for pathways gene.zscores <- qnorm(gene.pvalues, lower.tail = FALSE) gst <- t(sapply(annotation, pathwayStats, gene.zscores, minSize=5, path.method)) diff --git a/R/geneStats.R b/R/geneStats.R index 2d296dc..c821f85 100644 --- a/R/geneStats.R +++ b/R/geneStats.R @@ -1,4 +1,4 @@ -#' Gene Level Statistics +#' Molecule Level Statistics #' #' Takes a vector of statistics with each element corresponds to a treatment vs control comparison, #' and calculates a combined statistics accross multiple treatments. diff --git a/R/kinasePA.R b/R/kinasePA.R new file mode 100644 index 0000000..3042609 --- /dev/null +++ b/R/kinasePA.R @@ -0,0 +1,59 @@ +#' Direction Analysis for Kinases +#' +#' This is a wrapper for runing directPA for kinase perturbation analysis (kinasePA) +#' +#' @usage kinasePA(Tc, direction, annotation, minSize=5, substrate.method="OSP", +#' kinase.method="Stouffer", visualize=TRUE, ...) +#' +#' @param Tc a numeric matrix. The columns are phosphorylation sites and the columns are treatments vs +#' control statistics. +#' @param direction the direction to be tested for enrichment. Either specified as a degree for +#' two-dimensional analysis or as contrast (in a triplet) for three-dimensional analysis. +#' @param annotation a list with names correspond to kinases and elements correspond to substrates belong +#' to each kinase, respectively. +#' @param minSize the size of annotation groups to be considered for calculating enrichment. Groups +#' that are smaller than the minSize will be removed from the analysis. +#' @param substrate.method the method to be used for integrating statistics across treatments for each +#' substrate (phosphorylation site). Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is OSP. +#' @param kinase.method the method to be used for integrating statistics of all phosphorylation +#' sites that belongs to a kinase. Available methods are Stouffer, OSP, Fisher, and maxP. +#' Default method is Stouffer. +#' @param visualize whether to visualize the plot. +#' @param ... other visualization parameters to pass on. +#' @return a list that contains directional p-values for each substrate and directional enrichment for +#' each kinase. +#' @export +#' @examples +#' +#' # load the phosphoproteomics dataset +#' data(HEK) +#' +#' # load the kinase-substrate annoations +#' data(PhosphoSite) +#' +#' # direction pathway analysis in 2-dimensional space. Implemented as rotating by degree +#' # (1) test combined effect of Torin1 and Rapamycin vs insul both on "down-regulation" +#' # (180 degree to original direction) +#' kPA <- kinasePA(Tc=HEK, direction=pi, annotation=PhosphoSite.mouse) +#' kPA$kinase[order(unlist(kPA$kinase[,1])),][1:20,] +#' # rank substrates on the direciton of interest +#' sort(kPA$substrate.pvalues)[1:20] +#' +#' # (2) test combined effect of Torin1 and Rapamycin vs insul on "no change and down-regulation" +#' # (135 degree to the original direction) +#' kPA <- kinasePA(Tc=HEK, direction=pi*3/4, annotation=PhosphoSite.mouse) +#' kPA$kinase[order(unlist(kPA$kinase[,1])),][1:20,] +#' +#' # (3) test combined effect of Torin1 and Rapamycin vs insul on "down-regulation and no change" +#' # (225 degree to the original direction) +#' kPA <- kinasePA(Tc=HEK, direction=pi*5/4, annotation=PhosphoSite.mouse) +#' kPA$kinase[order(unlist(kPA$kinase[,1])),][1:20,] +#' +kinasePA <- function(Tc, direction, annotation, minSize=5, substrate.method="OSP", kinase.method="Stouffer", visualize=TRUE, ...){ + dPA <- directPA(Tc, direction, annotation, minSize, gene.method=substrate.method, path.method=kinase.method, visualize, ...) + kPA <- list() + kPA$substrate.pvalues <- dPA$gene.pvalues + kPA$kinase <- dPA$gst + return(kPA) +} diff --git a/R/perturbPlot2d.R b/R/perturbPlot2d.R index b46dcfb..7e20d7b 100644 --- a/R/perturbPlot2d.R +++ b/R/perturbPlot2d.R @@ -4,7 +4,7 @@ #' annotation list such as pathway annotation or kinase-substrate annotation, and visualize the enrichment #' of pathways or kinases in direction specific manner. #' -#' @usage directEnrichPlot2d(Tc, annotation, minSize=5, ...) +#' @usage perturbPlot2d(Tc, annotation, minSize=5, ...) #' @param Tc a numeric matrix. The columns are genes or phosphorylation sites and the columns are treatments #' vs control statistics. #' @param annotation a list with names correspond to pathways or kinases and elements correspond to genes or diff --git a/README.md b/README.md index 551fb49..2f136ba 100644 --- a/README.md +++ b/README.md @@ -2,7 +2,7 @@ ======== #### Description -`directPA' is a package designed for analyzing (1) gene/protein expression data using microarray, RNA-seq, or proteomics data, and (2) kinase enrichment using phosphoproteomics data. It intergrates multiple experimental treatments and test the effect of each combination of treatments using a directional approach. +`directPA' is a package designed to identify combinatorial effects of multiple treatments and/or perturbations on pathways and kinases profiled by microarray, RNA-seq, proteomics, or phosphoproteomics data. #### Download and install The relsease version can be downloaded from CRAN [link](http://cran.r-project.org/web/packages/directPA/); diff --git a/directPA_1.0.tar.gz b/directPA_1.0.tar.gz deleted file mode 100644 index 245ec98f7245b2a72f7c84782a8aa76a66ccce0d..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 98092 zcmV(#K;*w4iwFQ{7-muc1MK}%m~G9oF9@$%waT__+qP}nwr#Jnty#9UR@t0o+qV1N z`+s`(>F)12UtgTF`=a-6K4WB#h{%Y@86zWeT!pEXvzdvjq7cK6|8-#e2OMl{|JE{b zurvPqGZQQ8zqkJlKbV-AS(sS=`LX@OXZlBs{Rbi2|3aVs18y#^M$UwUKb)-`ZOy#? zM{;J)F8>)0{pZ5;e~-Vskerx`qL7H#|9l(&|KiWY!St{Av#_(XuyXtpe->tDwjYFy 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