<- params$Condition_1
Condition_1 <- params$Condition_2 Condition_2
Top GO Analysis for GC Inhibitor
library(RNASeqBulkExploratory)
library(DT)
library(ggplot2)
library(AnnotationDbi)
## Loading required package: stats4
## Loading required package: BiocGenerics
##
## Attaching package: 'BiocGenerics'
## The following objects are masked from 'package:stats':
##
## IQR, mad, sd, var, xtabs
## The following objects are masked from 'package:base':
##
## anyDuplicated, aperm, append, as.data.frame, basename, cbind,
## colnames, dirname, do.call, duplicated, eval, evalq, Filter, Find,
## get, grep, grepl, intersect, is.unsorted, lapply, Map, mapply,
## match, mget, order, paste, pmax, pmax.int, pmin, pmin.int,
## Position, rank, rbind, Reduce, rownames, sapply, setdiff, sort,
## table, tapply, union, unique, unsplit, which.max, which.min
## Loading required package: Biobase
## Welcome to Bioconductor
##
## Vignettes contain introductory material; view with
## 'browseVignettes()'. To cite Bioconductor, see
## 'citation("Biobase")', and for packages 'citation("pkgname")'.
## Loading required package: IRanges
## Loading required package: S4Vectors
##
## Attaching package: 'S4Vectors'
## The following objects are masked from 'package:base':
##
## expand.grid, I, unname
library(org.Hs.eg.db)
##
library(gridExtra)
##
## Attaching package: 'gridExtra'
## The following object is masked from 'package:Biobase':
##
## combine
## The following object is masked from 'package:BiocGenerics':
##
## combine
library(RColorBrewer)
library(viridis)
## Loading required package: viridisLite
library(topGO)
## Loading required package: graph
## Loading required package: GO.db
##
## Loading required package: SparseM
##
## Attaching package: 'SparseM'
## The following object is masked from 'package:base':
##
## backsolve
##
## groupGOTerms: GOBPTerm, GOMFTerm, GOCCTerm environments built.
##
## Attaching package: 'topGO'
## The following object is masked from 'package:IRanges':
##
## members
library(data.table)
##
## Attaching package: 'data.table'
## The following object is masked from 'package:IRanges':
##
## shift
## The following objects are masked from 'package:S4Vectors':
##
## first, second
library(tidyr)
##
## Attaching package: 'tidyr'
## The following object is masked from 'package:S4Vectors':
##
## expand
library(dplyr)
##
## Attaching package: 'dplyr'
## The following objects are masked from 'package:data.table':
##
## between, first, last
## The following object is masked from 'package:graph':
##
## union
## The following object is masked from 'package:gridExtra':
##
## combine
## The following object is masked from 'package:AnnotationDbi':
##
## select
## The following objects are masked from 'package:IRanges':
##
## collapse, desc, intersect, setdiff, slice, union
## The following objects are masked from 'package:S4Vectors':
##
## first, intersect, rename, setdiff, setequal, union
## The following object is masked from 'package:Biobase':
##
## combine
## The following objects are masked from 'package:BiocGenerics':
##
## combine, intersect, setdiff, union
## The following objects are masked from 'package:stats':
##
## filter, lag
## The following objects are masked from 'package:base':
##
## intersect, setdiff, setequal, union
library(SummarizedExperiment)
## Loading required package: MatrixGenerics
## Loading required package: matrixStats
##
## Attaching package: 'matrixStats'
## The following object is masked from 'package:dplyr':
##
## count
## The following objects are masked from 'package:Biobase':
##
## anyMissing, rowMedians
##
## Attaching package: 'MatrixGenerics'
## The following objects are masked from 'package:matrixStats':
##
## colAlls, colAnyNAs, colAnys, colAvgsPerRowSet, colCollapse,
## colCounts, colCummaxs, colCummins, colCumprods, colCumsums,
## colDiffs, colIQRDiffs, colIQRs, colLogSumExps, colMadDiffs,
## colMads, colMaxs, colMeans2, colMedians, colMins, colOrderStats,
## colProds, colQuantiles, colRanges, colRanks, colSdDiffs, colSds,
## colSums2, colTabulates, colVarDiffs, colVars, colWeightedMads,
## colWeightedMeans, colWeightedMedians, colWeightedSds,
## colWeightedVars, rowAlls, rowAnyNAs, rowAnys, rowAvgsPerColSet,
## rowCollapse, rowCounts, rowCummaxs, rowCummins, rowCumprods,
## rowCumsums, rowDiffs, rowIQRDiffs, rowIQRs, rowLogSumExps,
## rowMadDiffs, rowMads, rowMaxs, rowMeans2, rowMedians, rowMins,
## rowOrderStats, rowProds, rowQuantiles, rowRanges, rowRanks,
## rowSdDiffs, rowSds, rowSums2, rowTabulates, rowVarDiffs, rowVars,
## rowWeightedMads, rowWeightedMeans, rowWeightedMedians,
## rowWeightedSds, rowWeightedVars
## The following object is masked from 'package:Biobase':
##
## rowMedians
## Loading required package: GenomicRanges
## Loading required package: GenomeInfoDb
library(sechm)
source("../../plotGenesInTerm_v2.R")
<- params$Dataset
Dataset <- params$logFcTh
logFcTh <- params$FdrTh
FdrTh <- ifelse(is.null(params$OutputFolder), getwd(), params$OutputFolder)
OutputFolder
if (dir.exists(OutputFolder) == FALSE) {
dir.create(OutputFolder, recursive=TRUE)
}
# List with differential expression results
<- readRDS(params$DEAFile)
DEA
#SE object coming from DEA, but not containing specific contrast results
<- readRDS(params$SEFile) SE_DEA
if(! identical(rownames(SE_DEA), row.names(DEA[[Condition_1]][[Condition_2]]$Res))){
stop('Expression data in SE and results from differential espression analysis are inconsistent.')
}## Loading required package: DESeq2
rowData(SE_DEA) <- cbind(rowData(SE_DEA)[,1:6], DEA[[Condition_1]][[Condition_2]]$Res)
# Column names must be set to be compliant with the required format to be recognized by ORA
names(rowData(SE_DEA))[which(names(rowData(SE_DEA))=='log2FoldChange')] <- 'logFC'
names(rowData(SE_DEA))[which(names(rowData(SE_DEA))=='padj')] <- 'FDR'
#metadata(SE_DEA_Prel)$annotation <- 'hsa'
14850 genes in 20 samples have been testes for differential expression.
Imposing a threshold of 1 on the Log2FC and 0.01 on the FDR (as specified in parameters), 351 genes are selected: 48 up-regulated genes and 303 down-regulated genes.
plotVolcanoSE(SE=SE_DEA, FdrTh=FdrTh, logFcTh=logFcTh, FdrCeil=1e-10, logFcCeil=4)
Heatmaps for DEGs, showing scaled vst values.
<- dplyr::filter(data.frame(rowData(SE_DEA)), FDR < FdrTh & abs(logFC) > logFcTh)
DEGs
<- c('darkblue', "purple","white","lightgoldenrod1", 'goldenrod1')
ScaledCols
colData(SE_DEA)$Condition <- factor(colData(SE_DEA)$Condition, levels=c("CTL", "DMSO", "AhHyd_Ag", "AhHyd_Inh", "Andr_Ag", "Andr_Inh", "Estr_Ag", "Estr_Inh", "GC_Ag", "GC_Inh", "LivX_Ag", "LivX_Inh", "Ret_Ag", "Ret_Inh", "Thyr_Ag", "Thyr_Inh" ))
metadata(SE_DEA)$anno_colors <- list(Condition = c('DMSO' = 'grey30', 'CTL' = 'azure3',
'AhHyd_Ag'='#F8766D', 'AhHyd_Inh'='#F8766D50',
'Andr_Ag'='#fccb17', 'Andr_Inh'='#C49A0050',
"Estr_Ag"= '#53B400', "Estr_Inh"= '#53B40050',
'GC_Ag' = '#00C094', 'GC_Inh' = '#00C09450',
'LivX_Ag' = '#00B6EB', 'LivX_Inh' = '#00B6EB50',
'Ret_Ag' = '#A58AFF', 'Ret_Inh' = '#A58AFF50',
'Thyr_Ag' = '#FB61D7', 'Thyr_Inh' = '#FB61D750'
SeqRun = c("20210310" = "orange", "20210724" = "green4", "20220422" =
), "#66ACB7"))
sechm(SE_DEA, features=DEGs$GeneName, assayName="vst", gaps_at="Condition", show_rownames=FALSE,
top_annotation=c('Condition', 'SeqRun'), hmcols=ScaledCols, show_colnames=TRUE,
do.scale=TRUE, breaks=0.85, column_title = "Scaled Vst Values")
Gene ontology enrichment analysis is performed on the set of 351 genes using TopGO with Fisher statistics and weight01 algorithm.
For each specified domain of the ontology:
<- topGOGeneVectors(SE_DEA, FdrTh=FdrTh, logFcTh=logFcTh) GeneVectors
## Gene vector contains levels: 0,1
## Gene vector contains levels: 0,1
## Gene vector contains levels: 0,1
Therefore:
<- ifelse(length(grep('BP', params$TopGO))!=0, TRUE, FALSE)
BpEval <- ifelse(length(grep('MF', params$TopGO))!=0, TRUE, FALSE)
MfEval <- ifelse(length(grep('CC', params$TopGO))!=0, TRUE, FALSE) CcEval
On the basis of the analysis settings, the enrichment for Biological Process IS performed.
<- topGO::annFUN.org(whichOnto="BP", feasibleGenes=names(GeneVectors$DEGenes),
BPann mapping="org.Hs.eg.db", ID="symbol") %>% inverseList()
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=BPann, ontology='BP',
ResBPAll desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=paste0(OutputFolder), fileName='BPAll')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 11591 GO terms found. )
##
## Build GO DAG topology ..........
## ( 15045 GO terms and 33988 relations. )
##
## Annotating nodes ...............
## ( 13128 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 3282 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 18: 1 nodes to be scored (0 eliminated genes)
##
## Level 17: 3 nodes to be scored (0 eliminated genes)
##
## Level 16: 6 nodes to be scored (18 eliminated genes)
##
## Level 15: 10 nodes to be scored (50 eliminated genes)
##
## Level 14: 25 nodes to be scored (130 eliminated genes)
##
## Level 13: 56 nodes to be scored (284 eliminated genes)
##
## Level 12: 76 nodes to be scored (1007 eliminated genes)
##
## Level 11: 172 nodes to be scored (2937 eliminated genes)
##
## Level 10: 291 nodes to be scored (4590 eliminated genes)
##
## Level 9: 427 nodes to be scored (6068 eliminated genes)
##
## Level 8: 485 nodes to be scored (7723 eliminated genes)
##
## Level 7: 578 nodes to be scored (9952 eliminated genes)
##
## Level 6: 522 nodes to be scored (11318 eliminated genes)
##
## Level 5: 336 nodes to be scored (12180 eliminated genes)
##
## Level 4: 191 nodes to be scored (12645 eliminated genes)
##
## Level 3: 85 nodes to be scored (12854 eliminated genes)
##
## Level 2: 17 nodes to be scored (12955 eliminated genes)
##
## Level 1: 1 nodes to be scored (13002 eliminated genes)
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=BPann, ontology='BP',
ResBPDown desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=paste0(OutputFolder), fileName='BPDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 11591 GO terms found. )
##
## Build GO DAG topology ..........
## ( 15045 GO terms and 33988 relations. )
##
## Annotating nodes ...............
## ( 13128 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 3176 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 18: 1 nodes to be scored (0 eliminated genes)
##
## Level 17: 3 nodes to be scored (0 eliminated genes)
##
## Level 16: 6 nodes to be scored (18 eliminated genes)
##
## Level 15: 10 nodes to be scored (50 eliminated genes)
##
## Level 14: 25 nodes to be scored (130 eliminated genes)
##
## Level 13: 51 nodes to be scored (284 eliminated genes)
##
## Level 12: 72 nodes to be scored (1007 eliminated genes)
##
## Level 11: 159 nodes to be scored (2817 eliminated genes)
##
## Level 10: 273 nodes to be scored (4351 eliminated genes)
##
## Level 9: 411 nodes to be scored (5809 eliminated genes)
##
## Level 8: 465 nodes to be scored (7461 eliminated genes)
##
## Level 7: 561 nodes to be scored (9859 eliminated genes)
##
## Level 6: 515 nodes to be scored (11236 eliminated genes)
##
## Level 5: 333 nodes to be scored (12170 eliminated genes)
##
## Level 4: 188 nodes to be scored (12643 eliminated genes)
##
## Level 3: 85 nodes to be scored (12853 eliminated genes)
##
## Level 2: 17 nodes to be scored (12955 eliminated genes)
##
## Level 1: 1 nodes to be scored (13002 eliminated genes)
GOTable(ResBPDown$ResSel, maxGO=20)
<- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=BPann, ontology='BP',
ResBPUp desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='BPUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 11591 GO terms found. )
##
## Build GO DAG topology ..........
## ( 15045 GO terms and 33988 relations. )
##
## Annotating nodes ...............
## ( 13128 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 941 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 15: 1 nodes to be scored (0 eliminated genes)
##
## Level 14: 2 nodes to be scored (0 eliminated genes)
##
## Level 13: 12 nodes to be scored (49 eliminated genes)
##
## Level 12: 21 nodes to be scored (260 eliminated genes)
##
## Level 11: 40 nodes to be scored (2060 eliminated genes)
##
## Level 10: 67 nodes to be scored (3704 eliminated genes)
##
## Level 9: 76 nodes to be scored (4503 eliminated genes)
##
## Level 8: 99 nodes to be scored (5535 eliminated genes)
##
## Level 7: 142 nodes to be scored (6790 eliminated genes)
##
## Level 6: 167 nodes to be scored (8995 eliminated genes)
##
## Level 5: 155 nodes to be scored (10771 eliminated genes)
##
## Level 4: 96 nodes to be scored (12185 eliminated genes)
##
## Level 3: 49 nodes to be scored (12748 eliminated genes)
##
## Level 2: 13 nodes to be scored (12895 eliminated genes)
##
## Level 1: 1 nodes to be scored (12990 eliminated genes)
GOTable(ResBPUp$ResSel, maxGO=20)
topGOBarplotAll(TopGOResAll=ResBPAll$ResSel, TopGOResDown=ResBPDown$ResSel, TopGOResUp=ResBPUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
plotGenesInTerm_v2(ResBPAll$ResSel, ResBPAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
plotGenesInTerm_v2(ResBPDown$ResSel, ResBPDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
plotGenesInTerm_v2(ResBPUp$ResSel, ResBPUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
On the basis of the analysis settings, the enrichment for Molecular Function IS performed.
<- topGO::annFUN.org(whichOnto='MF', feasibleGenes=names(GeneVectors$DEGenes),
MFann mapping='org.Hs.eg.db', ID='symbol') %>% inverseList()
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=MFann, ontology='MF',
ResMFAll desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFAll')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4114 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4579 GO terms and 5967 relations. )
##
## Annotating nodes ...............
## ( 13476 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 465 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 5 nodes to be scored (0 eliminated genes)
##
## Level 10: 7 nodes to be scored (0 eliminated genes)
##
## Level 9: 16 nodes to be scored (109 eliminated genes)
##
## Level 8: 24 nodes to be scored (1163 eliminated genes)
##
## Level 7: 45 nodes to be scored (3066 eliminated genes)
##
## Level 6: 80 nodes to be scored (3491 eliminated genes)
##
## Level 5: 117 nodes to be scored (4721 eliminated genes)
##
## Level 4: 116 nodes to be scored (7779 eliminated genes)
##
## Level 3: 43 nodes to be scored (10553 eliminated genes)
##
## Level 2: 11 nodes to be scored (11502 eliminated genes)
##
## Level 1: 1 nodes to be scored (13345 eliminated genes)
<- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=MFann, ontology='MF',
ResMFDown desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4114 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4579 GO terms and 5967 relations. )
##
## Annotating nodes ...............
## ( 13476 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 445 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 4 nodes to be scored (0 eliminated genes)
##
## Level 10: 7 nodes to be scored (0 eliminated genes)
##
## Level 9: 15 nodes to be scored (90 eliminated genes)
##
## Level 8: 24 nodes to be scored (1163 eliminated genes)
##
## Level 7: 42 nodes to be scored (3064 eliminated genes)
##
## Level 6: 75 nodes to be scored (3491 eliminated genes)
##
## Level 5: 114 nodes to be scored (4704 eliminated genes)
##
## Level 4: 112 nodes to be scored (7693 eliminated genes)
##
## Level 3: 41 nodes to be scored (10521 eliminated genes)
##
## Level 2: 10 nodes to be scored (11443 eliminated genes)
##
## Level 1: 1 nodes to be scored (13345 eliminated genes)
GOTable(ResMFDown$ResSel, maxGO=20)
<- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=MFann, ontology='MF',
ResMFUp desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4114 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4579 GO terms and 5967 relations. )
##
## Annotating nodes ...............
## ( 13476 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 187 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 2 nodes to be scored (0 eliminated genes)
##
## Level 10: 2 nodes to be scored (0 eliminated genes)
##
## Level 9: 9 nodes to be scored (39 eliminated genes)
##
## Level 8: 10 nodes to be scored (1037 eliminated genes)
##
## Level 7: 15 nodes to be scored (2669 eliminated genes)
##
## Level 6: 25 nodes to be scored (2980 eliminated genes)
##
## Level 5: 38 nodes to be scored (4046 eliminated genes)
##
## Level 4: 49 nodes to be scored (5437 eliminated genes)
##
## Level 3: 26 nodes to be scored (8796 eliminated genes)
##
## Level 2: 10 nodes to be scored (10249 eliminated genes)
##
## Level 1: 1 nodes to be scored (13286 eliminated genes)
GOTable(ResMFUp$ResSel, maxGO=20)
topGOBarplotAll(TopGOResAll=ResMFAll$ResSel, TopGOResDown=ResMFDown$ResSel, TopGOResUp=ResMFUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
plotGenesInTerm_v2(ResMFAll$ResSel, ResMFAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
plotGenesInTerm_v2(ResMFDown$ResSel, ResMFDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
plotGenesInTerm_v2(ResMFUp$ResSel, ResMFUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
On the basis of the analysis settings, the enrichment for Cellular Component IS performed.
<- topGO::annFUN.org(whichOnto='CC', feasibleGenes=names(GeneVectors$DEGenes),
CCann mapping='org.Hs.eg.db', ID='symbol') %>% inverseList()
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=CCann, ontology='CC',
ResCCAll desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCAll')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1740 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1936 GO terms and 3272 relations. )
##
## Annotating nodes ...............
## ( 13716 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 396 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 18 nodes to be scored (0 eliminated genes)
##
## Level 10: 36 nodes to be scored (0 eliminated genes)
##
## Level 9: 48 nodes to be scored (644 eliminated genes)
##
## Level 8: 58 nodes to be scored (2133 eliminated genes)
##
## Level 7: 64 nodes to be scored (4361 eliminated genes)
##
## Level 6: 56 nodes to be scored (8245 eliminated genes)
##
## Level 5: 47 nodes to be scored (10016 eliminated genes)
##
## Level 4: 32 nodes to be scored (11954 eliminated genes)
##
## Level 3: 34 nodes to be scored (13136 eliminated genes)
##
## Level 2: 2 nodes to be scored (13552 eliminated genes)
##
## Level 1: 1 nodes to be scored (13659 eliminated genes)
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=CCann, ontology='CC',
ResCCDown desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1740 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1936 GO terms and 3272 relations. )
##
## Annotating nodes ...............
## ( 13716 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 378 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 17 nodes to be scored (0 eliminated genes)
##
## Level 10: 35 nodes to be scored (0 eliminated genes)
##
## Level 9: 48 nodes to be scored (625 eliminated genes)
##
## Level 8: 54 nodes to be scored (2132 eliminated genes)
##
## Level 7: 60 nodes to be scored (4361 eliminated genes)
##
## Level 6: 54 nodes to be scored (8219 eliminated genes)
##
## Level 5: 46 nodes to be scored (9996 eliminated genes)
##
## Level 4: 29 nodes to be scored (11950 eliminated genes)
##
## Level 3: 32 nodes to be scored (13136 eliminated genes)
##
## Level 2: 2 nodes to be scored (13551 eliminated genes)
##
## Level 1: 1 nodes to be scored (13659 eliminated genes)
GOTable(ResCCDown$ResSel, maxGO=20)
# Wrapper function for topGO analysis
<- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=CCann, ontology='CC',
ResCCUp desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1740 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1936 GO terms and 3272 relations. )
##
## Annotating nodes ...............
## ( 13716 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 159 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 2 nodes to be scored (0 eliminated genes)
##
## Level 10: 6 nodes to be scored (0 eliminated genes)
##
## Level 9: 8 nodes to be scored (93 eliminated genes)
##
## Level 8: 19 nodes to be scored (833 eliminated genes)
##
## Level 7: 24 nodes to be scored (1873 eliminated genes)
##
## Level 6: 28 nodes to be scored (6533 eliminated genes)
##
## Level 5: 27 nodes to be scored (8889 eliminated genes)
##
## Level 4: 20 nodes to be scored (11736 eliminated genes)
##
## Level 3: 22 nodes to be scored (12998 eliminated genes)
##
## Level 2: 2 nodes to be scored (13504 eliminated genes)
##
## Level 1: 1 nodes to be scored (13633 eliminated genes)
GOTable(ResCCUp$ResSel, maxGO=20)
topGOBarplotAll(TopGOResAll=ResCCAll$ResSel, TopGOResDown=ResCCDown$ResSel, TopGOResUp=ResCCUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
plotGenesInTerm_v2(ResCCAll$ResSel, ResCCAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
plotGenesInTerm_v2(ResCCDown$ResSel, ResCCDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
plotGenesInTerm_v2(ResCCUp$ResSel, ResCCUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
<- sessionInfo()
SessionInfo <- date()
Date #
save.image(paste0(OutputFolder, Dataset, 'FunctionalAnalysisWorkspace.RData'))
SessionInfo
## R version 4.2.1 (2022-06-23)
## Platform: x86_64-pc-linux-gnu (64-bit)
## Running under: Ubuntu 20.04.4 LTS
##
## Matrix products: default
## BLAS: /usr/lib/x86_64-linux-gnu/blas/libblas.so.3.9.0
## LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.9.0
##
## locale:
## [1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C
## [3] LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8
## [5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8
## [7] LC_PAPER=en_US.UTF-8 LC_NAME=C
## [9] LC_ADDRESS=C LC_TELEPHONE=C
## [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C
##
## attached base packages:
## [1] stats4 stats graphics grDevices utils datasets methods
## [8] base
##
## other attached packages:
## [1] DESeq2_1.38.3 sechm_1.6.0
## [3] SummarizedExperiment_1.28.0 GenomicRanges_1.50.2
## [5] GenomeInfoDb_1.34.9 MatrixGenerics_1.10.0
## [7] matrixStats_0.63.0 dplyr_1.1.0
## [9] tidyr_1.3.0 data.table_1.14.8
## [11] topGO_2.50.0 SparseM_1.81
## [13] GO.db_3.16.0 graph_1.76.0
## [15] viridis_0.6.2 viridisLite_0.4.1
## [17] RColorBrewer_1.1-3 gridExtra_2.3
## [19] org.Hs.eg.db_3.16.0 AnnotationDbi_1.60.0
## [21] IRanges_2.32.0 S4Vectors_0.36.1
## [23] Biobase_2.58.0 BiocGenerics_0.44.0
## [25] ggplot2_3.4.1 DT_0.27
## [27] RNASeqBulkExploratory_0.2.1
##
## loaded via a namespace (and not attached):
## [1] Rtsne_0.16 colorspace_2.1-0 rjson_0.2.21
## [4] ellipsis_0.3.2 circlize_0.4.15 XVector_0.38.0
## [7] GlobalOptions_0.1.2 clue_0.3-64 rstudioapi_0.14
## [10] farver_2.1.1 bit64_4.0.5 fansi_1.0.4
## [13] codetools_0.2-19 doParallel_1.0.17 cachem_1.0.7
## [16] geneplotter_1.76.0 knitr_1.42 jsonlite_1.8.4
## [19] annotate_1.76.0 cluster_2.1.4 png_0.1-8
## [22] compiler_4.2.1 httr_1.4.5 lazyeval_0.2.2
## [25] Matrix_1.5-3 fastmap_1.1.1 cli_3.6.1
## [28] htmltools_0.5.4 tools_4.2.1 gtable_0.3.1
## [31] glue_1.6.2 GenomeInfoDbData_1.2.9 V8_4.2.2
## [34] Rcpp_1.0.10 jquerylib_0.1.4 vctrs_0.6.2
## [37] Biostrings_2.66.0 iterators_1.0.14 crosstalk_1.2.0
## [40] xfun_0.37 stringr_1.5.0 lifecycle_1.0.3
## [43] XML_3.99-0.13 ca_0.71.1 zlibbioc_1.44.0
## [46] scales_1.2.1 TSP_1.2-2 parallel_4.2.1
## [49] ComplexHeatmap_2.14.0 yaml_2.3.7 curl_5.0.0
## [52] memoise_2.0.1 sass_0.4.5 stringi_1.7.12
## [55] RSQLite_2.3.0 highr_0.10 randomcoloR_1.1.0.1
## [58] foreach_1.5.2 seriation_1.4.1 BiocParallel_1.32.5
## [61] shape_1.4.6 rlang_1.1.1 pkgconfig_2.0.3
## [64] bitops_1.0-7 evaluate_0.20 lattice_0.20-45
## [67] purrr_1.0.1 labeling_0.4.2 htmlwidgets_1.6.1
## [70] bit_4.0.5 tidyselect_1.2.0 magrittr_2.0.3
## [73] R6_2.5.1 generics_0.1.3 DelayedArray_0.24.0
## [76] DBI_1.1.3 pillar_1.8.1 withr_2.5.0
## [79] KEGGREST_1.38.0 RCurl_1.98-1.10 tibble_3.2.1
## [82] crayon_1.5.2 utf8_1.2.3 plotly_4.10.1
## [85] rmarkdown_2.20 GetoptLong_1.0.5 locfit_1.5-9.7
## [88] grid_4.2.1 blob_1.2.3 digest_0.6.31
## [91] xtable_1.8-4 munsell_0.5.0 registry_0.5-1
## [94] bslib_0.4.2
Date
## [1] "Fri Jul 18 18:30:16 2025"