Mercurial > repos > vandelj > giant_aptsummarize
comparison src/VolcanoPlotsScript.R @ 0:708f43bda2b6 draft
"planemo upload for repository https://github.com/juliechevalier/GIANT/tree/master commit cb276a594444c8f32e9819fefde3a21f121d35df"
| author | vandelj |
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| date | Fri, 26 Jun 2020 09:35:11 -0400 |
| parents | |
| children |
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| -1:000000000000 | 0:708f43bda2b6 |
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| 1 # R script to plot volcanos through Galaxy based GIANT tool | |
| 2 # written by Jimmy Vandel | |
| 3 # | |
| 4 # | |
| 5 initial.options <- commandArgs(trailingOnly = FALSE) | |
| 6 file.arg.name <- "--file=" | |
| 7 script.name <- sub(file.arg.name, "", initial.options[grep(file.arg.name, initial.options)]) | |
| 8 script.basename <- dirname(script.name) | |
| 9 source(file.path(script.basename, "utils.R")) | |
| 10 source(file.path(script.basename, "getopt.R")) | |
| 11 | |
| 12 #addComment("Welcome R!") | |
| 13 | |
| 14 # setup R error handling to go to stderr | |
| 15 options( show.error.messages=F, error = function () { cat(geterrmessage(), file=stderr() ); q( "no", 1, F ) } ) | |
| 16 | |
| 17 # we need that to not crash galaxy with an UTF8 error on German LC settings. | |
| 18 loc <- Sys.setlocale("LC_MESSAGES", "en_US.UTF-8") | |
| 19 loc <- Sys.setlocale("LC_NUMERIC", "C") | |
| 20 | |
| 21 #get starting time | |
| 22 start.time <- Sys.time() | |
| 23 | |
| 24 options(stringAsfactors = FALSE, useFancyQuotes = FALSE) | |
| 25 args <- commandArgs() | |
| 26 | |
| 27 # get options, using the spec as defined by the enclosed list. | |
| 28 # we read the options from the default: commandArgs(TRUE). | |
| 29 spec <- matrix(c( | |
| 30 "statisticsFile", "i", 1, "character", | |
| 31 "volcanoName" , "n", 1, "character", | |
| 32 "pvalColumnName" , "p", 1, "character", | |
| 33 "fdrColumnName" , "m", 1, "character", | |
| 34 "fcColumnName" , "c", 1, "character", | |
| 35 "fcKind","d", 1, "character", | |
| 36 "fdrThreshold","s", 1, "double", | |
| 37 "fcThreshold","e", 1, "double", | |
| 38 "organismID","x",1,"character", | |
| 39 "rowNameType","y",1,"character", | |
| 40 "log", "l", 1, "character", | |
| 41 "outputFile" , "o", 1, "character", | |
| 42 "format", "f", 1, "character", | |
| 43 "quiet", "q", 0, "logical"), | |
| 44 byrow=TRUE, ncol=4) | |
| 45 opt <- getopt(spec) | |
| 46 | |
| 47 # enforce the following required arguments | |
| 48 if (is.null(opt$log)) { | |
| 49 addComment("[ERROR]'log file' is required\n") | |
| 50 q( "no", 1, F ) | |
| 51 } | |
| 52 addComment("[INFO]Start of R script",T,opt$log,display=FALSE) | |
| 53 if (is.null(opt$statisticsFile)) { | |
| 54 addComment("[ERROR]'statisticsFile' is required",T,opt$log) | |
| 55 q( "no", 1, F ) | |
| 56 } | |
| 57 if (length(opt$pvalColumnName)==0 || length(opt$fdrColumnName)==0 || length(opt$fcColumnName)==0) { | |
| 58 addComment("[ERROR]no selected columns",T,opt$log) | |
| 59 q( "no", 1, F ) | |
| 60 } | |
| 61 if (length(opt$pvalColumnName)!=length(opt$fcColumnName) || length(opt$pvalColumnName)!=length(opt$fdrColumnName)) { | |
| 62 addComment("[ERROR]different number of selected columns between p.val, adj-p.val and FC ",T,opt$log) | |
| 63 q( "no", 1, F ) | |
| 64 } | |
| 65 if (is.null(opt$fcKind)) { | |
| 66 addComment("[ERROR]'fcKind' is required",T,opt$log) | |
| 67 q( "no", 1, F ) | |
| 68 } | |
| 69 if (is.null(opt$fdrThreshold)) { | |
| 70 addComment("[ERROR]'FDR threshold' is required",T,opt$log) | |
| 71 q( "no", 1, F ) | |
| 72 } | |
| 73 if (is.null(opt$fcThreshold)) { | |
| 74 addComment("[ERROR]'FC threshold' is required",T,opt$log) | |
| 75 q( "no", 1, F ) | |
| 76 } | |
| 77 if (is.null(opt$outputFile)) { | |
| 78 addComment("[ERROR]'output file' is required",T,opt$log) | |
| 79 q( "no", 1, F ) | |
| 80 } | |
| 81 if (is.null(opt$format)) { | |
| 82 addComment("[ERROR]'output format' is required",T,opt$log) | |
| 83 q( "no", 1, F ) | |
| 84 } | |
| 85 | |
| 86 #demande si le script sera bavard | |
| 87 verbose <- if (is.null(opt$quiet)) { | |
| 88 TRUE | |
| 89 }else{ | |
| 90 FALSE | |
| 91 } | |
| 92 | |
| 93 #paramètres internes | |
| 94 addComment("[INFO]Parameters checked test mode !",T,opt$log,display=FALSE) | |
| 95 | |
| 96 addComment(c("[INFO]Working directory: ",getwd()),TRUE,opt$log,display=FALSE) | |
| 97 addComment(c("[INFO]Command line: ",args),TRUE,opt$log,display=FALSE) | |
| 98 | |
| 99 #directory for plots | |
| 100 dir.create(file.path(getwd(), "plotDir")) | |
| 101 dir.create(file.path(getwd(), "plotLyDir")) | |
| 102 | |
| 103 #charge des packages silencieusement | |
| 104 suppressPackageStartupMessages({ | |
| 105 library("methods") | |
| 106 library("biomaRt") | |
| 107 library("ggplot2") | |
| 108 library("plotly") | |
| 109 library("stringr") | |
| 110 }) | |
| 111 | |
| 112 #define some usefull variable | |
| 113 nbVolcanosToPlot=length(opt$pvalColumnName) | |
| 114 | |
| 115 #load input file | |
| 116 statDataMatrix=read.csv(file=file.path(getwd(), opt$statisticsFile),header=F,sep="\t",colClasses="character") | |
| 117 #remove first colum to convert it as rownames | |
| 118 rownames(statDataMatrix)=statDataMatrix[,1] | |
| 119 statDataMatrix=statDataMatrix[,-1] | |
| 120 | |
| 121 #identify lines without adjusted p-value info (should contain the same content as rownames) and replace them with NA values | |
| 122 FDRinfo=rep(TRUE,nbVolcanosToPlot) | |
| 123 for(iVolcano in 1:nbVolcanosToPlot){ | |
| 124 #input parameter should be None when adjusted p-val are not available | |
| 125 if(opt$fdrColumnName[iVolcano]=="None"){ | |
| 126 #content of the corresponding column should also be the same as rownames | |
| 127 if(!all(statDataMatrix[,(iVolcano-1)*3+2]==rownames(statDataMatrix))){ | |
| 128 addComment(c("[ERROR]It seems that input stat matrix contains adjusted p-values for volcano",iVolcano,"whereas input parameter indicates that not."),T,opt$log) | |
| 129 q( "no", 1, F ) | |
| 130 } | |
| 131 FDRinfo[iVolcano]=FALSE | |
| 132 statDataMatrix[,(iVolcano-1)*3+2]=NA | |
| 133 } | |
| 134 } | |
| 135 | |
| 136 if(is.data.frame(statDataMatrix)){ | |
| 137 statDataMatrix=data.matrix(statDataMatrix) | |
| 138 }else{ | |
| 139 statDataMatrix=data.matrix(as.numeric(statDataMatrix)) | |
| 140 } | |
| 141 | |
| 142 #check if available column number match with volcano requested number | |
| 143 if(ncol(statDataMatrix)!=3*nbVolcanosToPlot){ | |
| 144 addComment("[ERROR]Input file column number is different from requested volcano number",T,opt$log) | |
| 145 q( "no", 1, F ) | |
| 146 } | |
| 147 | |
| 148 #build global dataFrame with data and fill with p.val and log2(FC) and FDR | |
| 149 dataFrame=data.frame(row.names = rownames(statDataMatrix)) | |
| 150 #start with p-value | |
| 151 dataFrame$p.value=statDataMatrix[,seq(1,nbVolcanosToPlot*3,3),drop=FALSE] | |
| 152 #compute FDR if needed or just get available info | |
| 153 dataFrame$adj_p.value=dataFrame$p.value | |
| 154 for(iVolcano in 1:nbVolcanosToPlot){ | |
| 155 #adjusted p-value are already computed | |
| 156 if(FDRinfo[iVolcano]){ | |
| 157 dataFrame$adj_p.value[,iVolcano]=statDataMatrix[,(iVolcano-1)*3+2,drop=FALSE] | |
| 158 }else{ | |
| 159 #adjusted p-value should be computed based on p-val using FDR | |
| 160 dataFrame$adj_p.value[,iVolcano]=p.adjust(dataFrame$p.value[,iVolcano,drop=FALSE],"fdr") | |
| 161 addComment(c("[INFO]Adjusted p-values are not available in input for volcano",iVolcano,", FDR approach will be used on available raw p-values"),T,opt$log) | |
| 162 } | |
| 163 } | |
| 164 if(opt$fcKind=="FC"){ | |
| 165 #we should transform as Log2FC | |
| 166 dataFrame$coefficients=log2(statDataMatrix[,seq(3,nbVolcanosToPlot*3,3),drop=FALSE]) | |
| 167 addComment(c("[INFO]FC are converted in log2(FC) for plotting"),T,opt$log) | |
| 168 }else{ | |
| 169 dataFrame$coefficients=statDataMatrix[,seq(3,nbVolcanosToPlot*3,3),drop=FALSE] | |
| 170 } | |
| 171 | |
| 172 addComment(c("[INFO]Input data available for",nbVolcanosToPlot,"volcano(s) with",nrow(statDataMatrix),"rows"),T,opt$log) | |
| 173 | |
| 174 | |
| 175 #plot VOLCANOs | |
| 176 volcanoPerPage=1 | |
| 177 logFCthreshold=log2(opt$fcThreshold) | |
| 178 iToPlot=1 | |
| 179 plotVector=list() | |
| 180 volcanoNameList=c() | |
| 181 for (iVolcano in 1:nbVolcanosToPlot){ | |
| 182 | |
| 183 if(nchar(opt$volcanoName[iVolcano])>0){ | |
| 184 curentVolcanoName=opt$volcanoName[iVolcano] | |
| 185 }else{ | |
| 186 curentVolcanoName=paste(iVolcano,opt$pvalColumnName[iVolcano],sep="_") | |
| 187 } | |
| 188 | |
| 189 #keep only rows without NA for p-val, adjusted p-val and coeff | |
| 190 pValToPlot=dataFrame$p.value[,iVolcano] | |
| 191 fdrToPlot=dataFrame$adj_p.value[,iVolcano] | |
| 192 coeffToPlot=dataFrame$coefficients[,iVolcano] | |
| 193 | |
| 194 rowToRemove=unique(c(which(is.na(pValToPlot)),which(is.na(fdrToPlot)),which(is.na(coeffToPlot)))) | |
| 195 if(length(rowToRemove)>0){ | |
| 196 pValToPlot=pValToPlot[-rowToRemove] | |
| 197 fdrToPlot=fdrToPlot[-rowToRemove] | |
| 198 coeffToPlot=coeffToPlot[-rowToRemove] | |
| 199 } | |
| 200 addComment(c("[INFO]For",curentVolcanoName,"volcano,",length(rowToRemove),"rows are discarded due to NA values,",length(pValToPlot),"remaining rows."),T,opt$log) | |
| 201 | |
| 202 #save volcano name | |
| 203 volcanoNameList=c(volcanoNameList,curentVolcanoName) | |
| 204 | |
| 205 #remove characters possibly troubling | |
| 206 volcanoFileName=iVolcano | |
| 207 | |
| 208 #define the log10(p-val) threshold corresponding to FDR threshold fixed by user | |
| 209 probeWithLowFDR=-log10(pValToPlot[which(fdrToPlot<=opt$fdrThreshold)]) | |
| 210 pvalThresholdFDR=NULL | |
| 211 if(length(probeWithLowFDR)>0)pvalThresholdFDR=min(probeWithLowFDR) | |
| 212 | |
| 213 #get significant points over FC and FDR thresholds | |
| 214 significativePoints=intersect(which(abs(coeffToPlot)>=logFCthreshold),which(fdrToPlot<=opt$fdrThreshold)) | |
| 215 | |
| 216 #to reduce size of html plot, we keep 20000 points maximum sampled amongst genes with pval>=33%(pval) and abs(log2(FC))<=66%(abs(log2(FC))) | |
| 217 htmlPointsToRemove=intersect(which(abs(coeffToPlot)<=quantile(abs(coeffToPlot),c(0.66))),which(pValToPlot>=quantile(abs(pValToPlot),c(0.33)))) | |
| 218 if(length(htmlPointsToRemove)>20000){ | |
| 219 htmlPointsToRemove=setdiff(htmlPointsToRemove,sample(htmlPointsToRemove,20000)) | |
| 220 }else{ | |
| 221 htmlPointsToRemove=c() | |
| 222 } | |
| 223 | |
| 224 xMinLimPlot=min(coeffToPlot)-0.2 | |
| 225 xMaxLimPlot=max(coeffToPlot)+0.2 | |
| 226 yMaxLimPlot= max(-log10(pValToPlot))+0.2 | |
| 227 | |
| 228 if(length(significativePoints)>0){ | |
| 229 dataSignifToPlot=data.frame(pval=-log10(pValToPlot[significativePoints]),FC=coeffToPlot[significativePoints],description=paste(names(coeffToPlot[significativePoints]),"\n","FC: " , round(2^coeffToPlot[significativePoints],2) , " | Adjusted p-val: ",prettyNum(fdrToPlot[significativePoints],digits=4), sep="")) | |
| 230 #to test if remains any normal points to draw | |
| 231 if(length(significativePoints)<length(pValToPlot)){ | |
| 232 dataToPlot=data.frame(pval=-log10(pValToPlot[-significativePoints]),FC=coeffToPlot[-significativePoints],description=paste("FC: " , round(2^coeffToPlot[-significativePoints],2) , " | Adjusted p-val: ",prettyNum(fdrToPlot[-significativePoints],digits=4), sep="")) | |
| 233 }else{ | |
| 234 dataToPlot=data.frame(pval=0,FC=0,description="null") | |
| 235 } | |
| 236 }else{ | |
| 237 dataToPlot=data.frame(pval=-log10(pValToPlot),FC=coeffToPlot,description=paste("FC: " , round(2^coeffToPlot,2) , " | Adjusted p-val: ",prettyNum(fdrToPlot,digits=4), sep="")) | |
| 238 } | |
| 239 | |
| 240 ##traditional plot | |
| 241 | |
| 242 p <- ggplot(data=dataToPlot, aes(x=FC, y=pval)) + geom_point() + | |
| 243 theme_bw() + ggtitle(curentVolcanoName) + ylab(label="-Log10(p-val)") + xlab(label="Log2 Fold Change") + | |
| 244 theme(panel.border=element_blank(),plot.title = element_text(hjust = 0.5),legend.position="none") | |
| 245 if(logFCthreshold!=0) p <- p + geom_vline(xintercept=-logFCthreshold, color="salmon",linetype="dotted", size=1) + geom_vline(xintercept=logFCthreshold, color="salmon",linetype="dotted", size=1) + geom_text(data.frame(text=c(paste(c("log2(1/FC=",opt$fcThreshold,")"),collapse=""),paste(c("log2(FC=",opt$fcThreshold,")"),collapse="")),x=c(-logFCthreshold,logFCthreshold),y=c(0,0)),mapping=aes(x=x, y=y, label=text), size=4, angle=90, vjust=-0.4, hjust=0, color="salmon") | |
| 246 if(!is.null(pvalThresholdFDR)) p <- p + geom_hline(yintercept=pvalThresholdFDR, color="skyblue1",linetype="dotted", size=0.5) + geom_text(data.frame(text=c(paste(c("Adjusted pval limit(",opt$fdrThreshold,")"),collapse="")),x=c(xMinLimPlot),y=c(pvalThresholdFDR)),mapping=aes(x=x, y=y, label=text), size=4, vjust=0, hjust=0, color="skyblue3") | |
| 247 if(length(significativePoints)>0)p <- p + geom_point(data=dataSignifToPlot,aes(colour=description)) | |
| 248 | |
| 249 ##interactive plot | |
| 250 | |
| 251 if(length(htmlPointsToRemove)>0){ | |
| 252 pointToRemove=union(htmlPointsToRemove,significativePoints) | |
| 253 #to test if it remains any normal points to draw | |
| 254 if(length(pointToRemove)<length(pValToPlot)){ | |
| 255 dataToPlot=data.frame(pval=-log10(pValToPlot[-pointToRemove]),FC=coeffToPlot[-pointToRemove],description=paste("FC: " , round(2^coeffToPlot[-pointToRemove],2) , " | Adjusted p-val: ", prettyNum(fdrToPlot[-pointToRemove],digits=4), sep="")) | |
| 256 }else{ | |
| 257 dataToPlot=data.frame(pval=0,FC=0,description="null") | |
| 258 } | |
| 259 } | |
| 260 | |
| 261 if((nrow(dataToPlot)+length(significativePoints))>40000)addComment(c("[WARNING]For",curentVolcanoName,"volcano, numerous points to plot(",nrow(dataToPlot)+nrow(dataSignifToPlot),"), resulting volcano could be heavy, using more stringent thresholds could be helpful."),T,opt$log) | |
| 262 | |
| 263 phtml <- plot_ly(data=dataToPlot, x=~FC, y=~pval,type="scatter", mode="markers",showlegend = FALSE, marker = list(color="gray",opacity=0.5), text=~description, hoverinfo="text") %>% | |
| 264 layout(title = curentVolcanoName[iVolcano],xaxis=list(title="Log2 Fold Change",showgrid=TRUE, zeroline=FALSE),yaxis=list(title="-Log10(p-val)", showgrid=TRUE, zeroline=FALSE)) | |
| 265 if(length(significativePoints)>0) phtml=add_markers(phtml,data=dataSignifToPlot, x=~FC, y=~pval, mode="markers" , marker=list( color=log10(abs(dataSignifToPlot$FC)*dataSignifToPlot$pval),colorscale='Rainbow'), text=~description, hoverinfo="text", inherit = FALSE) %>% hide_colorbar() | |
| 266 if(logFCthreshold!=0){ | |
| 267 phtml=add_trace(phtml,x=c(-logFCthreshold,-logFCthreshold), y=c(0,yMaxLimPlot), type="scatter", mode = "lines", line=list(color="coral",dash="dash"), hoverinfo='none', showlegend = FALSE,inherit = FALSE) | |
| 268 phtml=add_annotations(phtml,x=-logFCthreshold,y=0,xref = "x",yref = "y",text = paste(c("log2(1/FC=",opt$fcThreshold,")"),collapse=""),xanchor = 'right',showarrow = F,textangle=270,font=list(color="coral")) | |
| 269 phtml=add_trace(phtml,x=c(logFCthreshold,logFCthreshold), y=c(0, yMaxLimPlot), type="scatter", mode = "lines", line=list(color="coral",dash="dash"), hoverinfo='none', showlegend = FALSE,inherit = FALSE) | |
| 270 phtml=add_annotations(phtml,x=logFCthreshold,y=0,xref = "x",yref = "y",text = paste(c("log2(FC=",opt$fcThreshold,")"),collapse=""),xanchor = 'right',showarrow = F,textangle=270,font=list(color="coral")) | |
| 271 } | |
| 272 if(!is.null(pvalThresholdFDR)){ | |
| 273 phtml=add_trace(phtml,x=c(xMinLimPlot,xMaxLimPlot), y=c(pvalThresholdFDR,pvalThresholdFDR), type="scatter", mode = "lines", line=list(color="cornflowerblue",dash="dash"), hoverinfo='none', showlegend = FALSE,inherit = FALSE) | |
| 274 phtml=add_annotations(phtml,x=xMinLimPlot,y=pvalThresholdFDR+0.1,xref = "x",yref = "y",text = paste(c("Adjusted pval limit(",opt$fdrThreshold,")"),collapse=""),xanchor = 'left',showarrow = F,font=list(color="cornflowerblue")) | |
| 275 } | |
| 276 plotVector[[length(plotVector)+1]]=p | |
| 277 | |
| 278 #save plotly files | |
| 279 pp <- ggplotly(phtml) | |
| 280 htmlwidgets::saveWidget(as_widget(pp), paste(c(file.path(getwd(), "plotLyDir"),"/Volcanos_",volcanoFileName,".html"),collapse=""),selfcontained = F) | |
| 281 | |
| 282 | |
| 283 if(iVolcano==nbVolcanosToPlot || length(plotVector)==volcanoPerPage){ | |
| 284 #plot and close the actual plot | |
| 285 if(opt$format=="pdf"){ | |
| 286 pdf(paste(c("./plotDir/Volcanos_",volcanoFileName,".pdf"),collapse=""))}else{ | |
| 287 png(paste(c("./plotDir/Volcanos_",volcanoFileName,".png"),collapse="")) | |
| 288 } | |
| 289 multiplot(plotlist=plotVector,cols=1) | |
| 290 dev.off() | |
| 291 if(iVolcano<nbVolcanosToPlot){ | |
| 292 #prepare for a new ploting file if necessary | |
| 293 plotVector=list() | |
| 294 iToPlot=iToPlot+1 | |
| 295 } | |
| 296 } | |
| 297 } | |
| 298 remove(dataToPlot,dataSignifToPlot) | |
| 299 addComment("[INFO]Volcanos drawn",T,opt$log,T,display=FALSE) | |
| 300 | |
| 301 | |
| 302 #now add anotation infos about genes | |
| 303 | |
| 304 rowItemInfo=NULL | |
| 305 if(!is.null(opt$rowNameType) && !is.null(opt$organismID)){ | |
| 306 ##get gene information from BioMart | |
| 307 #if(!require("biomaRt")){ | |
| 308 # source("https://bioconductor.org/biocLite.R") | |
| 309 # biocLite("biomaRt") | |
| 310 #} | |
| 311 | |
| 312 ensembl_hs_mart <- useMart(biomart="ensembl", dataset=opt$organismID) | |
| 313 ensembl_df <- getBM(attributes=c(opt$rowNameType,"description"),mart=ensembl_hs_mart) | |
| 314 rowItemInfo=ensembl_df[which(ensembl_df[,1]!=""),2] | |
| 315 rowItemInfo=unlist(lapply(rowItemInfo,function(x)substr(unlist(strsplit(x," \\[Source"))[1],1,30))) | |
| 316 names(rowItemInfo)=ensembl_df[which(ensembl_df[,1]!=""),1] | |
| 317 } | |
| 318 | |
| 319 #filter out genes with higher p-values for all comparisons | |
| 320 genesToKeep=names(which(apply(dataFrame$adj_p.value,1,function(x)length(which(x<=opt$fdrThreshold))>0))) | |
| 321 #filter out genes with lower FC for all comparisons | |
| 322 genesToKeep=intersect(genesToKeep,names(which(apply(dataFrame$coefficients,1,function(x)length(which(abs(x)>=logFCthreshold))>0)))) | |
| 323 | |
| 324 if(length(genesToKeep)>0){ | |
| 325 dataFrameNew=data.frame(row.names=genesToKeep) | |
| 326 | |
| 327 dataFrameNew$adj_p.value=matrix(dataFrame$adj_p.value[genesToKeep,,drop=FALSE],ncol=ncol(dataFrame$adj_p.value)) | |
| 328 rownames(dataFrameNew$adj_p.value)=genesToKeep | |
| 329 colnames(dataFrameNew$adj_p.value)=colnames(dataFrame$p.value) | |
| 330 | |
| 331 dataFrameNew$p.value=matrix(dataFrame$p.value[genesToKeep,,drop=FALSE],ncol=ncol(dataFrame$p.value)) | |
| 332 rownames(dataFrameNew$p.value)=genesToKeep | |
| 333 colnames(dataFrameNew$p.value)=colnames(dataFrame$adj_p.value) | |
| 334 | |
| 335 dataFrameNew$coefficients=matrix(dataFrame$coefficients[genesToKeep,,drop=FALSE],ncol=ncol(dataFrame$coefficients)) | |
| 336 rownames(dataFrameNew$coefficients)=genesToKeep | |
| 337 colnames(dataFrameNew$coefficients)=colnames(dataFrame$adj_p.value) | |
| 338 | |
| 339 dataFrame=dataFrameNew | |
| 340 rm(dataFrameNew) | |
| 341 }else{ | |
| 342 addComment("[WARNING]No significative genes",T,opt$log,display=FALSE) | |
| 343 } | |
| 344 | |
| 345 addComment("[INFO]Significant genes filtering done",T,opt$log,T,display=FALSE) | |
| 346 | |
| 347 | |
| 348 #plot VennDiagramm for genes below threshold between comparisons | |
| 349 #t=apply(dataFrame$adj_p.value[,1:4],2,function(x)names(which(x<=opt$threshold))) | |
| 350 #get.venn.partitions(t) | |
| 351 #vennCounts(dataFrame$adj_p.value[,1:4]<=opt$threshold) | |
| 352 | |
| 353 #make a simple sort genes based only on the first comparison | |
| 354 #newOrder=order(dataFrame$adj_p.value[,1]) | |
| 355 #dataFrame$adj_p.value=dataFrame$adj_p.value[newOrder,] | |
| 356 | |
| 357 #alternative sorting strategy based on the mean gene rank over all comparisons | |
| 358 if(length(genesToKeep)>1){ | |
| 359 currentRank=rep(0,nrow(dataFrame$adj_p.value)) | |
| 360 for(iVolcano in 1:ncol(dataFrame$adj_p.value)){ | |
| 361 currentRank=currentRank+rank(dataFrame$adj_p.value[,iVolcano]) | |
| 362 } | |
| 363 currentRank=currentRank/ncol(dataFrame$adj_p.value) | |
| 364 newOrder=order(currentRank) | |
| 365 rownames(dataFrame)=rownames(dataFrame)[newOrder] | |
| 366 | |
| 367 dataFrame$adj_p.value=matrix(dataFrame$adj_p.value[newOrder,],ncol=ncol(dataFrame$adj_p.value)) | |
| 368 rownames(dataFrame$adj_p.value)=rownames(dataFrame$p.value)[newOrder] | |
| 369 colnames(dataFrame$adj_p.value)=colnames(dataFrame$p.value) | |
| 370 | |
| 371 dataFrame$p.value=matrix(dataFrame$p.value[newOrder,],ncol=ncol(dataFrame$p.value)) | |
| 372 rownames(dataFrame$p.value)=rownames(dataFrame$adj_p.value) | |
| 373 colnames(dataFrame$p.value)=colnames(dataFrame$adj_p.value) | |
| 374 | |
| 375 dataFrame$coefficients=matrix(dataFrame$coefficients[newOrder,],ncol=ncol(dataFrame$coefficients)) | |
| 376 rownames(dataFrame$coefficients)=rownames(dataFrame$adj_p.value) | |
| 377 colnames(dataFrame$coefficients)=colnames(dataFrame$adj_p.value) | |
| 378 } | |
| 379 | |
| 380 #formating output matrix depending on genes to keep | |
| 381 if(length(genesToKeep)==0){ | |
| 382 outputData=matrix(0,ncol=ncol(dataFrame$adj_p.value)*4+2,nrow=3) | |
| 383 outputData[1,]=c("X","X",rep(volcanoNameList,each=4)) | |
| 384 outputData[2,]=c("X","X",rep(c("p-val","Adjusted.p-val","FC","log2(FC)"),ncol(dataFrame$adj_p.value))) | |
| 385 outputData[,1]=c("Volcano","Gene","noGene") | |
| 386 outputData[,2]=c("Comparison","Info","noInfo") | |
| 387 }else{ | |
| 388 if(length(genesToKeep)==1){ | |
| 389 outputData=matrix(0,ncol=ncol(dataFrame$adj_p.value)*4+2,nrow=3) | |
| 390 outputData[1,]=c("X","X",rep(volcanoNameList,each=4)) | |
| 391 outputData[2,]=c("X","X",rep(c("p-val","Adjusted.p-val","FC","log2(FC)"),ncol(dataFrame$adj_p.value))) | |
| 392 outputData[,1]=c("Volcano","Gene",genesToKeep) | |
| 393 outputData[,2]=c("Comparison","Info","na") | |
| 394 if(!is.null(rowItemInfo))outputData[3,2]=rowItemInfo[genesToKeep] | |
| 395 outputData[3,seq(3,ncol(outputData),4)]=prettyNum(dataFrame$p.value,digits=4) | |
| 396 outputData[3,seq(4,ncol(outputData),4)]=prettyNum(dataFrame$adj_p.value,digits=4) | |
| 397 outputData[3,seq(5,ncol(outputData),4)]=prettyNum(2^dataFrame$coefficients,digits=4) | |
| 398 outputData[3,seq(6,ncol(outputData),4)]=prettyNum(dataFrame$coefficients,digits=4) | |
| 399 }else{ | |
| 400 #format matrix to be correctly read by galaxy (move headers in first column and row) | |
| 401 outputData=matrix(0,ncol=ncol(dataFrame$adj_p.value)*4+2,nrow=nrow(dataFrame$adj_p.value)+2) | |
| 402 outputData[1,]=c("X","X",rep(volcanoNameList,each=4)) | |
| 403 outputData[2,]=c("X","X",rep(c("p-val","Adjusted.p-val","FC","log2(FC)"),ncol(dataFrame$adj_p.value))) | |
| 404 outputData[,1]=c("Volcano","Gene",rownames(dataFrame$adj_p.value)) | |
| 405 outputData[,2]=c("Comparison","Info",rep("na",nrow(dataFrame$adj_p.value))) | |
| 406 if(!is.null(rowItemInfo))outputData[3:nrow(outputData),2]=rowItemInfo[rownames(dataFrame$adj_p.value)] | |
| 407 outputData[3:nrow(outputData),seq(3,ncol(outputData),4)]=prettyNum(dataFrame$p.value,digits=4) | |
| 408 outputData[3:nrow(outputData),seq(4,ncol(outputData),4)]=prettyNum(dataFrame$adj_p.value,digits=4) | |
| 409 outputData[3:nrow(outputData),seq(5,ncol(outputData),4)]=prettyNum(2^dataFrame$coefficients,digits=4) | |
| 410 outputData[3:nrow(outputData),seq(6,ncol(outputData),4)]=prettyNum(dataFrame$coefficients,digits=4) | |
| 411 } | |
| 412 } | |
| 413 addComment("[INFO]Formated output",T,opt$log,display=FALSE) | |
| 414 | |
| 415 #write output results | |
| 416 write.table(outputData,file=opt$outputFile,quote=FALSE,sep="\t",col.names = F,row.names = F) | |
| 417 | |
| 418 | |
| 419 end.time <- Sys.time() | |
| 420 addComment(c("[INFO]Total execution time for R script:",as.numeric(end.time - start.time,units="mins"),"mins"),T,opt$log,display=FALSE) | |
| 421 | |
| 422 addComment("[INFO]End of R script",T,opt$log,display=FALSE) | |
| 423 | |
| 424 printSessionInfo(opt$log) | |
| 425 | |
| 426 #sessionInfo() |
