--- title: 【迁移】oligo：GEO/ArrayExpress芯片数据处理 urlname: oligo-GEO-ArrayExpress-xin-pian-shu-ju-chu-li date: 2022-07-29 19:00:11 tags: ['oligo', 'GEO', 'ArrayExpress'] --- ## 安装依赖 ```bash conda clean -y -all conda create -n geo -c conda-forge r-base=4.1.3 conda activate geo conda install -c conda-forge r-tidyverse=1.3.1 -y conda install -c conda-forge r-irkernel -y conda install -c conda-forge r-r.utils -y conda install -c conda-forge r-rcolorbrewer -y Rscript -e "IRkernel::installspec(name='geo', displayname='r-geo')" conda install -c bioconda bioconductor-geoquery -y conda install -c bioconda bioconductor-oligo -y conda install -c bioconda bioconductor-affy -y conda install -c bioconda bioconductor-pd.hugene.1.0.st.v1 -y # read.celfiles 需要啥安装啥 conda install -c bioconda bioconductor-pd.hg.u133a.2 -y conda install -c bioconda bioconductor-arrayexpress -y conda install -c bioconda bioconductor-biomart -y conda install -c bioconda bioconductor-affycoretools -y ``` ## 下载芯片数据 ### GEO ```R library(GEOquery) library(oligo) library(affy) gse="GSE43332" rawdata <- getGEOSuppFiles(gse) #下载原始数据 rawdata setwd(gse) untar("GSE43332_RAW.tar",exdir = ".") celfiles <- list.files(pattern = "*CEL.gz$") #批量查找并列出后缀为.gz的文件 data.raw <- read.celfiles(celfiles) ``` ### ArrayExpress * ~/dev/xray/xray -c ~/etc/xui2.json & * wget -e "https\_proxy=http://127.0.0.1:20809" https://www.ebi.ac.uk/arrayexpress/files/E-MEXP-1422/E-MEXP-1422.raw.1.zip -O E-MEXP-1422.raw.1.zip * unzip E-MEXP-1422.raw.1.zip ```R library(ArrayExpress) library(oligo) library(affy) # Sys.setenv("http_proxy"="http://127.0.0.1:20809") # Sys.setenv("https_proxy"="http://127.0.0.1:20809") # mexp1422=getAE("E-MTAB-6128", type = 'raw', extract = T) celfiles <- list.files(pattern = "*.CEL$") #批量查找并列出后缀为CEL的文件 data.raw <- read.celfiles(celfiles) ``` ## RMA标准化数据 ```R sampleNames(data.raw) <- sapply(strsplit(sampleNames(data.raw),"_",fixed=T), "[",1) sampleNames(data.raw) fit1 <- fitProbeLevelModel(data.raw) image(fit1,type="weights",which=1,main="weights") image(fit1,type="residuals",which=1,main="Residuals") image(fit1,type="sign.residuals",which=1,main="Residuals.sign") data.eset <- oligo::rma(data.raw) data.exprs <- exprs(data.eset) library(RColorBrewer) display.brewer.all() if(ncol(data.eset)>8){ colors <- c(brewer.pal(8,"Set2"),brewer.pal(ncol(data.eset)-8,"Set1")) }else{ colors <- brewer.pal(ncol(data.eset),"Set2") } boxplot(data.exprs,col=colors,main="afterRMA") ``` ## 探针转Symbol ### 使用IQR方法进行重复值的选择 使用 rownames(df) <- rowNn 时经常会遇到rowNn中有重复值的情况，此时需要使用合适的策略来选择需要保留的那一列。下面这个函数默认保留IQR值(四分位距)最大的那一列。通过传入不同的select_func参数值，也可以改用其他的保留选择策略。如 mean 来保留算数平均值最大的一列，也可以传入自己定义的函数。 ```R f_rm_duplicated <- function(NameL, reverse=F){ tmp <- data.frame(table(NameL)) if(reverse){ tmp <- tmp$NameL[tmp$Freq > 1] }else{ tmp <- tmp$NameL[tmp$Freq == 1] } which(NameL %in% as.character(tmp)) } f_dedup_IQR <- function(df, rowNn, select_func='IQR'){ if(typeof(select_func) == 'character'){ select_func = get(select_func) } # 拆出无重复的数据，后续不进行处理 noDup <- f_rm_duplicated(rowNn) tmp <- rowNn[noDup] noDup <- df[noDup,] rownames(noDup) <- tmp # 拆除有重复的数据 Dup <- f_rm_duplicated(rowNn, T) rowNn <- rowNn[Dup] Dup <- df[Dup,] rownames(Dup) <- NULL # 处理重复的数据 lc_tmp = by(Dup, rowNn, function(x){rownames(x)[which.max(apply(X = x, FUN = select_func, MARGIN = 1))]}) lc_probes = as.integer(lc_tmp) Dup = Dup[lc_probes,] rownames(Dup) <- rowNn[lc_probes] # 合并数据并返回 return(rbind(noDup,Dup)) } ``` ### 探针转Symbol * data.raw@annotation 可以得到注释的包信息，如pd.hg.u133a.2 * 在[bioconductor](https://www.bioconductor.org/)上搜索pd.hg.u133a.2，可以知道其Platform为HG-U133A\_2 * 谷歌搜索 HG-U133A\_2 + affymetrix，可以找到affymetrix官网对应的页面 * 谷歌搜索 affymetrix "HG-U133A\_2" site:www.ncbi.nlm.nih.gov，随便进一个GSE页面，找到对应的Platforms，如 GPL571 \[HG-U133A\_2\] Affymetrix Human Genome U133A 2.0 Array，说明HG-U133A\_2对应GPL571 ```R require(GEOquery) f_getGPL <- function(lc_GPLN, lc_local = F){ options(stringsAsFactors = F) if (!file.exists(lc_GPLN)){dir.create(lc_GPLN)} if(lc_local){ gpl=read.table(file.path(lc_GPLN, lc_GPLN), header = TRUE,fill = T,sep = "\t", comment.char = "#", stringsAsFactors = FALSE, quote = "") return(gpl) }else{ return(Table(getGEO(lc_GPLN,destdir = lc_GPLN))) } } GPL <- 'GPL6244' GPL <- f_getGPL(GPL) GPL[['Symbol']] <- GPL$gene_assignment GPL <- GPL[c('ID','Symbol')] GPL[GPL$Symbol == '---', 'Symbol'] = '---//---' tmp <- strsplit(x = GPL$Symbol, split = '//') tmp <- lapply(tmp, FUN = function(x){x[2]}) GPL[['Symbol']] = unlist(tmp) GPL$Symbol[GPL$Symbol == '---'] = GPL$ID[GPL$Symbol == '---'] rownames(GPL) <- as.character(GPL$ID) GPL <- GPL[as.character(rownames(data.exprs)),] data.exprs <- f_dedup_IQR(data.exprs, GPL$Symbol) data.exprs ``` ## GPL与Platform对应表 | gpl | bioc_package | title | | -------- | ---------------------------- | ------------------------------------------------------------------------------------------------------- | | GPL32 | mgu74a | [MG_U74A] Affymetrix Murine Genome U74A Array | | GPL33 | mgu74b | [MG_U74B] Affymetrix Murine Genome U74B Array | | GPL34 | mgu74c | [MG_U74C] Affymetrix Murine Genome U74C Array | | GPL71 | ag | [AG] Affymetrix Arabidopsis Genome Array | | GPL72 | drosgenome1 | [DrosGenome1] Affymetrix Drosophila Genome Array | | GPL74 | hcg110 | [HC_G110] Affymetrix Human Cancer Array | | GPL75 | mu11ksuba | [Mu11KsubA] Affymetrix Murine 11K SubA Array | | GPL76 | mu11ksubb | [Mu11KsubB] Affymetrix Murine 11K SubB Array | | GPL77 | mu19ksuba | [Mu19KsubA] Affymetrix Murine 19K SubA Array | | GPL78 | mu19ksubb | [Mu19KsubB] Affymetrix Murine 19K SubB Array | | GPL79 | mu19ksubc | [Mu19KsubC] Affymetrix Murine 19K SubC Array | | GPL80 | hu6800 | [Hu6800] Affymetrix Human Full Length HuGeneFL Array | | GPL81 | mgu74av2 | [MG_U74Av2] Affymetrix Murine Genome U74A Version 2 Array | | GPL82 | mgu74bv2 | [MG_U74Bv2] Affymetrix Murine Genome U74B Version 2 Array | | GPL83 | mgu74cv2 | [MG_U74Cv2] Affymetrix Murine Genome U74 Version 2 Array | | GPL85 | rgu34a | [RG_U34A] Affymetrix Rat Genome U34 Array | | GPL86 | rgu34b | [RG_U34B] Affymetrix Rat Genome U34 Array | | GPL87 | rgu34c | [RG_U34C] Affymetrix Rat Genome U34 Array | | GPL88 | rnu34 | [RN_U34] Affymetrix Rat Neurobiology U34 Array | | GPL89 | rtu34 | [RT_U34] Affymetrix Rat Toxicology U34 Array | | GPL90 | ygs98 | [YG_S98] Affymetrix Yeast Genome S98 Array | | GPL91 | hgu95av2 | [HG_U95A] Affymetrix Human Genome U95A Array | | GPL92 | hgu95b | [HG_U95B] Affymetrix Human Genome U95B Array | | GPL93 | hgu95c | [HG_U95C] Affymetrix Human Genome U95C Array | | GPL94 | hgu95d | [HG_U95D] Affymetrix Human Genome U95D Array | | GPL95 | hgu95e | [HG_U95E] Affymetrix Human Genome U95E Array | | GPL96 | hgu133a | [HG-U133A] Affymetrix Human Genome U133A Array | | GPL97 | hgu133b | [HG-U133B] Affymetrix Human Genome U133B Array | | GPL98 | hu35ksuba | [Hu35KsubA] Affymetrix Human 35K SubA Array | | GPL99 | hu35ksubb | [Hu35KsubB] Affymetrix Human 35K SubB Array | | GPL100 | hu35ksubc | [Hu35KsubC] Affymetrix Human 35K SubC Array | | GPL101 | hu35ksubd | [Hu35KsubD] Affymetrix Human 35K SubD Array | | GPL198 | ath1121501 | [ATH1-121501] Affymetrix Arabidopsis ATH1 Genome Array | | GPL199 | ecoli2 | [Ecoli_ASv2] Affymetrix E. coli Antisense Genome Array | | GPL200 | celegans | [Celegans] Affymetrix C. elegans Genome Array | | GPL201 | hgfocus | [HG-Focus] Affymetrix Human HG-Focus Target Array | | GPL339 | moe430a | [MOE430A] Affymetrix Mouse Expression 430A Array | | GPL340 | mouse4302 | [MOE430B] Affymetrix Mouse Expression 430B Array | | GPL341 | rae230a | [RAE230A] Affymetrix Rat Expression 230A Array | | GPL342 | rae230b | [RAE230B] Affymetrix Rat Expression 230B Array | | GPL570 | hgu133plus2 | [HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array | | GPL571 | hgu133a2 | [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array | | GPL886 | hgug4111a | Agilent-011871 Human 1B Microarray G4111A (Feature Number version) | | GPL887 | hgug4110b | Agilent-012097 Human 1A Microarray (V2) G4110B (Feature Number version) | | GPL1261 | mouse430a2 | [Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array | | GPL1318 | xenopuslaevis | [Xenopus_laevis] Affymetrix Xenopus laevis Genome Array | | GPL1319 | zebrafish | [Zebrafish] Affymetrix Zebrafish Genome Array | | GPL1322 | drosophila2 | [Drosophila_2] Affymetrix Drosophila Genome 2.0 Array | | GPL1352 | u133x3p | [U133_X3P] Affymetrix Human X3P Array | | GPL1355 | rat2302 | [Rat230_2] Affymetrix Rat Genome 230 2.0 Array | | GPL1708 | hgug4112a | Agilent-012391 Whole Human Genome Oligo Microarray G4112A (Feature Number version) | | GPL2112 | bovine | [Bovine] Affymetrix Bovine Genome Array | | GPL2529 | yeast2 | [Yeast_2] Affymetrix Yeast Genome 2.0 Array | | GPL2891 | h20kcod | GE Healthcare/Amersham Biosciences CodeLink™ UniSet Human 20K I Bioarray | | GPL2898 | adme16cod | GE Healthcare/Amersham Biosciences CodeLink™ ADME Rat 16-Assay Bioarray | | GPL3154 | ecoli2 | [E_coli_2] Affymetrix E. coli Genome 2.0 Array | | GPL3213 | chicken | [Chicken] Affymetrix Chicken Genome Array | | GPL3533 | porcine | [Porcine] Affymetrix Porcine Genome Array | | GPL3738 | canine2 | [Canine_2] Affymetrix Canine Genome 2.0 Array | | GPL3921 | hthgu133a | [HT_HG-U133A] Affymetrix HT Human Genome U133A Array | | GPL3979 | canine | [Canine] Affymetrix Canine Genome 1.0 Array | | GPL4032 | | [Maize] Affymetrix Maize Genome Array | | GPL4191 | h10kcod | CodeLink UniSet Human I Bioarray | | GPL5188 | huex10sttranscriptcluster | [HuEx-1_0-st] Affymetrix Human Exon 1.0 ST Array [probe set (exon) version] | | GPL5689 | hgug4100a | Agilent Human 1 cDNA Microarray (G4100A) [layout C] | | GPL6097 | illuminaHumanv1 | Illumina human-6 v1.0 expression beadchip | | GPL6102 | illuminaHumanv2 | Illumina human-6 v2.0 expression beadchip | | GPL6244 | hugene10sttranscriptcluster | [HuGene-1_0-st] Affymetrix Human Gene 1.0 ST Array [transcript (gene) version] | | GPL6246 | mogene10sttranscriptcluster | [MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] | | GPL6885 | illuminaMousev2 | Illumina MouseRef-8 v2.0 expression beadchip | | GPL6947 | illuminaHumanv3 | Illumina HumanHT-12 V3.0 expression beadchip | | GPL8300 | hgu95av2 | [HG_U95Av2] Affymetrix Human Genome U95 Version 2 Array | | GPL8321 | mouse430a2 | [Mouse430A_2] Affymetrix Mouse Genome 430A 2.0 Array | | GPL8490 | IlluminaHumanMethylation27k | Illumina HumanMethylation27 BeadChip (HumanMethylation27_270596_v.1.2) | | GPL10558 | illuminaHumanv4 | Illumina HumanHT-12 V4.0 expression beadchip | | GPL11532 | hugene11sttranscriptcluster | [HuGene-1_1-st] Affymetrix Human Gene 1.1 ST Array [transcript (gene) version] | | GPL13497 | HsAgilentDesign026652 | Agilent-026652 Whole Human Genome Microarray 4x44K v2 (Probe Name version) | | GPL13534 | IlluminaHumanMethylation450k | Illumina HumanMethylation450 BeadChip (HumanMethylation450_15017482) | | GPL13667 | hgu219 | [HG-U219] Affymetrix Human Genome U219 Array | | GPL14877 | hgu133plus2 | Affymetrix Human Genome U133 Plus 2.0 Array [Brainarray Version 13, HGU133Plus2_Hs_ENTREZG] | | GPL15380 | GGHumanMethCancerPanelv1 | Illumina Sentrix Array Matrix (SAM) - GoldenGate Methylation Cancer Panel I | | GPL15396 | hthgu133b | [HT_HG-U133B] Affymetrix HT Human Genome U133B Array [custom CDF: ENTREZ brainarray v. 14] | | GPL17556 | hugene10sttranscriptcluster | [HuGene-1_0-st] Affymetrix Human Gene 1.0 ST Array [HuGene10stv1_Hs_ENTREZG_17.0.0] | | GPL17897 | hthgu133a | [HT_HG-U133A] Affymetrix Human Genome U133A Array (custom CDF: HTHGU133A_Hs_ENTREZG.cdf version 17.0.0) | | GPL18190 | hugene11sttranscriptcluster | [HuGene-1_1-st] Affymetrix Human Gene 1.1 ST Array [CDF: Brainarray HuGene11stv1_Hs_ENTREZG_15.1.0] |