TopGO - CTL04
Date: July 18, 2025
Condition_1 <- params$Condition_1
Condition_2 <- params$Condition_2Top GO Analysis for Ret AgvsInh
1. Environment Set Up
library(RNASeqBulkExploratory)
library(DT)
library(ggplot2)
library(AnnotationDbi)
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library(org.Hs.eg.db)
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library(RColorBrewer)
library(viridis)
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library(topGO)
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library(SummarizedExperiment)
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## Loading required package: GenomicRanges
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library(sechm)
source("../../plotGenesInTerm_v2.R")Dataset <- params$Dataset
logFcTh <- params$logFcTh
FdrTh <- params$FdrTh
OutputFolder <- ifelse(is.null(params$OutputFolder), getwd(), params$OutputFolder)
if (dir.exists(OutputFolder) == FALSE) {
dir.create(OutputFolder, recursive=TRUE)
}2. Data Upload
- Summarized Experiment object containing expression data used for DEA
and gene and sample metadata
- DEA object, containing results of the differential expression
2.1 Load Data from DEA
# List with differential expression results
DEA <- readRDS(params$DEAFile)
#SE object coming from DEA, but not containing specific contrast results
SE_DEA <- readRDS(params$SEFile)2.2 Add DEA results to SE
if(! identical(rownames(SE_DEA), row.names(DEA[[Condition_1]][[Condition_2]]$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'14305 genes in 21 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), 3541 genes are selected: 2171 up-regulated genes and 1370 down-regulated genes.
4. RESULTS VISUALIZATION
4.1 Volcano plot
The results of the differential expression analysis are visualized by Volcano plot. An interactive version is included in the html (only genes with FDR < threshold), while a static version is saved.
plotVolcanoSE(SE=SE_DEA, FdrTh=FdrTh, logFcTh=logFcTh, FdrCeil=1e-10, logFcCeil=4)4.3 Heatmap for significant genes
Heatmaps for DEGs, showing scaled vst values.
DEGs <- dplyr::filter(data.frame(rowData(SE_DEA)), FDR < FdrTh & abs(logFC) > logFcTh)
ScaledCols <- c('darkblue', "purple","white","lightgoldenrod1", 'goldenrod1')
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)$anno_colors <- list(Condition = c('DMSO' = 'grey30', 'CTL' = 'azure3',
'AhHyd_Ag'='#F8766D', 'AhHyd_Inh'='#F8766D50',
'Andr_Ag'='#fccb17', 'Andr_Inh'='#C49A0050',
"Estr_Ag"= '#53B400', "Estr_Inh"= '#53B40050',
'GC_Ag' = '#00C094', 'GC_Inh' = '#00C09450',
'LivX_Ag' = '#00B6EB', 'LivX_Inh' = '#00B6EB50',
'Ret_Ag' = '#A58AFF', 'Ret_Inh' = '#A58AFF50',
'Thyr_Ag' = '#FB61D7', 'Thyr_Inh' = '#FB61D750'
))
sechm(SE_DEA, features=DEGs$GeneName, assayName="vst", gaps_at="Condition", show_rownames=FALSE,
top_annotation=c('Condition'), hmcols=ScaledCols, show_colnames=TRUE,
do.scale=TRUE, breaks=0.85)## `use_raster` is automatically set to TRUE for a matrix with more than
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5. TOPGO for Gene Ontology Enrichment analysis
Gene ontology enrichment analysis is performed on the set of 3541 genes using TopGO with Fisher statistics and weight01 algorithm.
For each specified domain of the ontology:
- Enrichment analysis on all DEGs or splitted in down- and up-regulated
5.1 Selection of modulated genes and generation of gene vectors
I generate vectors for the gene universe, all modulated genes, up-regulated genes and down-regulated genes in the format required by TopGo.
GeneVectors <- topGOGeneVectors(SE_DEA, FdrTh=FdrTh, logFcTh=logFcTh)## Gene vector contains levels: 0,1
## Gene vector contains levels: 0,1
## Gene vector contains levels: 0,1Therefore:
- universe genes: 14305 genes
- modulated genes: 3541 genes
- down-regulated genes: 1370 genes of interest
- up-regulated genes: 2171 genes of interest
Then I set parameters according to the gene ontology domains to be evaluated. By default, Biological Process and Molecular Function domains are interrogated.
BpEval <- ifelse(length(grep('BP', params$TopGO))!=0, TRUE, FALSE)
MfEval <- ifelse(length(grep('MF', params$TopGO))!=0, TRUE, FALSE)
CcEval <- ifelse(length(grep('CC', params$TopGO))!=0, TRUE, FALSE)5.2 TopGO analysis: Biological Process
On the basis of the analysis settings, the enrichment for Biological Process IS performed.
Biological Process Analysis for ALL modulated genes: 3541 genes
BPann <- topGO::annFUN.org(whichOnto="BP", feasibleGenes=names(GeneVectors$DEGenes),
mapping="org.Hs.eg.db", ID="symbol") %>% inverseList()
# Wrapper function for topGO analysis
ResBPAll <- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=BPann, ontology='BP',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$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 .....
## ( 11416 GO terms found. )
##
## Build GO DAG topology ..........
## ( 14877 GO terms and 33562 relations. )
##
## Annotating nodes ...............
## ( 12665 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
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## the algorithm is scoring 4814 nontrivial nodes
## parameters:
## test statistic: fisher
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## Level 18: 1 nodes to be scored (0 eliminated genes)
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## Level 17: 4 nodes to be scored (0 eliminated genes)
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## Level 16: 12 nodes to be scored (18 eliminated genes)
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## Level 15: 25 nodes to be scored (73 eliminated genes)
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## Level 14: 56 nodes to be scored (235 eliminated genes)
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## Level 13: 91 nodes to be scored (533 eliminated genes)
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## Level 12: 156 nodes to be scored (1480 eliminated genes)
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## Level 11: 335 nodes to be scored (3347 eliminated genes)
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## Level 10: 535 nodes to be scored (5163 eliminated genes)
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## Level 9: 683 nodes to be scored (6805 eliminated genes)
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## Level 8: 736 nodes to be scored (8610 eliminated genes)
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## Level 7: 782 nodes to be scored (10222 eliminated genes)
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## Level 6: 656 nodes to be scored (11217 eliminated genes)
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## Level 5: 404 nodes to be scored (11894 eliminated genes)
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## Level 4: 226 nodes to be scored (12269 eliminated genes)
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## Level 3: 93 nodes to be scored (12434 eliminated genes)
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## Level 2: 18 nodes to be scored (12508 eliminated genes)
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## Level 1: 1 nodes to be scored (12546 eliminated genes)Biological Process Analysis for DOWN-REGULATED genes: 1370 genes
# Wrapper function for topGO analysis
ResBPDown <- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=BPann, ontology='BP',
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 .....
## ( 11416 GO terms found. )
##
## Build GO DAG topology ..........
## ( 14877 GO terms and 33562 relations. )
##
## Annotating nodes ...............
## ( 12665 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 4602 nontrivial nodes
## parameters:
## test statistic: fisher
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## Level 18: 1 nodes to be scored (0 eliminated genes)
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## Level 17: 4 nodes to be scored (0 eliminated genes)
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## Level 16: 10 nodes to be scored (18 eliminated genes)
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## Level 15: 22 nodes to be scored (73 eliminated genes)
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## Level 14: 49 nodes to be scored (214 eliminated genes)
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## Level 13: 79 nodes to be scored (503 eliminated genes)
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## Level 12: 145 nodes to be scored (1339 eliminated genes)
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## Level 11: 310 nodes to be scored (3196 eliminated genes)
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## Level 10: 501 nodes to be scored (5106 eliminated genes)
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## Level 9: 651 nodes to be scored (6667 eliminated genes)
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## Level 8: 704 nodes to be scored (8461 eliminated genes)
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## Level 7: 749 nodes to be scored (10159 eliminated genes)
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## Level 6: 643 nodes to be scored (11193 eliminated genes)
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## Level 5: 401 nodes to be scored (11870 eliminated genes)
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## Level 4: 221 nodes to be scored (12267 eliminated genes)
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## Level 3: 93 nodes to be scored (12434 eliminated genes)
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## Level 2: 18 nodes to be scored (12508 eliminated genes)
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## Level 1: 1 nodes to be scored (12546 eliminated genes)GOTable(ResBPDown$ResSel, maxGO=20)Biological Process Analysis for UP-REGULATED genes: 2171 genes
ResBPUp <- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=BPann, ontology='BP',
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 .....
## ( 11416 GO terms found. )
##
## Build GO DAG topology ..........
## ( 14877 GO terms and 33562 relations. )
##
## Annotating nodes ...............
## ( 12665 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 4603 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 18: 1 nodes to be scored (0 eliminated genes)
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## Level 17: 3 nodes to be scored (0 eliminated genes)
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## Level 16: 11 nodes to be scored (18 eliminated genes)
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## Level 15: 24 nodes to be scored (69 eliminated genes)
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## Level 14: 53 nodes to be scored (215 eliminated genes)
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## Level 13: 86 nodes to be scored (516 eliminated genes)
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## Level 12: 139 nodes to be scored (1445 eliminated genes)
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## Level 11: 303 nodes to be scored (3307 eliminated genes)
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## Level 10: 484 nodes to be scored (5076 eliminated genes)
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## Level 9: 645 nodes to be scored (6595 eliminated genes)
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## Level 8: 706 nodes to be scored (8433 eliminated genes)
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## Level 7: 768 nodes to be scored (10092 eliminated genes)
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## Level 6: 647 nodes to be scored (11181 eliminated genes)
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## Level 5: 399 nodes to be scored (11879 eliminated genes)
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## Level 4: 223 nodes to be scored (12266 eliminated genes)
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## Level 3: 92 nodes to be scored (12434 eliminated genes)
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## Level 2: 18 nodes to be scored (12508 eliminated genes)
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## Level 1: 1 nodes to be scored (12546 eliminated genes)GOTable(ResBPUp$ResSel, maxGO=20)Result visualization: Barplot
topGOBarplotAll(TopGOResAll=ResBPAll$ResSel, TopGOResDown=ResBPDown$ResSel, TopGOResUp=ResBPUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
Top Terms associated Genes
All
plotGenesInTerm_v2(ResBPAll$ResSel, ResBPAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
Down
plotGenesInTerm_v2(ResBPDown$ResSel, ResBPDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
Up
plotGenesInTerm_v2(ResBPUp$ResSel, ResBPUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
5.3 TopGO analysis: Molecular Function
On the basis of the analysis settings, the enrichment for Molecular Function IS performed.
Molecular Function Enrichment for ALL modulated genes: 3541 genes
MFann <- topGO::annFUN.org(whichOnto='MF', feasibleGenes=names(GeneVectors$DEGenes),
mapping='org.Hs.eg.db', ID='symbol') %>% inverseList()
# Wrapper function for topGO analysis
ResMFAll <- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=MFann, ontology='MF',
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 .....
## ( 4060 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4530 GO terms and 5903 relations. )
##
## Annotating nodes ...............
## ( 12999 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 817 nontrivial nodes
## parameters:
## test statistic: fisher
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## Level 11: 7 nodes to be scored (0 eliminated genes)
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## Level 10: 13 nodes to be scored (0 eliminated genes)
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## Level 9: 31 nodes to be scored (152 eliminated genes)
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## Level 8: 61 nodes to be scored (1213 eliminated genes)
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## Level 7: 110 nodes to be scored (3201 eliminated genes)
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## Level 6: 157 nodes to be scored (3858 eliminated genes)
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## Level 5: 186 nodes to be scored (5384 eliminated genes)
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## Level 4: 176 nodes to be scored (8297 eliminated genes)
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## Level 3: 58 nodes to be scored (10483 eliminated genes)
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## Level 2: 17 nodes to be scored (11240 eliminated genes)
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## Level 1: 1 nodes to be scored (12883 eliminated genes)Molecular Function Enrichment for DOWN-REGULATED genes: 1370 genes
ResMFDown <- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=MFann, ontology='MF',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4060 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4530 GO terms and 5903 relations. )
##
## Annotating nodes ...............
## ( 12999 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 748 nontrivial nodes
## parameters:
## test statistic: fisher
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## Level 11: 7 nodes to be scored (0 eliminated genes)
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## Level 10: 10 nodes to be scored (0 eliminated genes)
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## Level 9: 24 nodes to be scored (152 eliminated genes)
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## Level 8: 53 nodes to be scored (1165 eliminated genes)
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## Level 7: 103 nodes to be scored (3117 eliminated genes)
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## Level 6: 136 nodes to be scored (3784 eliminated genes)
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## Level 5: 174 nodes to be scored (5312 eliminated genes)
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## Level 4: 167 nodes to be scored (8146 eliminated genes)
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## Level 3: 56 nodes to be scored (10445 eliminated genes)
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## Level 2: 17 nodes to be scored (11224 eliminated genes)
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## Level 1: 1 nodes to be scored (12883 eliminated genes)GOTable(ResMFDown$ResSel, maxGO=20)Molecular Function Analysis for UP-REGULATED genes: 2171 genes
ResMFUp <- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=MFann, ontology='MF',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4060 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4530 GO terms and 5903 relations. )
##
## Annotating nodes ...............
## ( 12999 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 779 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 7 nodes to be scored (0 eliminated genes)
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## Level 10: 12 nodes to be scored (0 eliminated genes)
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## Level 9: 30 nodes to be scored (152 eliminated genes)
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## Level 8: 58 nodes to be scored (1187 eliminated genes)
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## Level 7: 97 nodes to be scored (3174 eliminated genes)
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## Level 6: 152 nodes to be scored (3810 eliminated genes)
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## Level 5: 180 nodes to be scored (5236 eliminated genes)
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## Level 4: 172 nodes to be scored (8241 eliminated genes)
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## Level 3: 54 nodes to be scored (10459 eliminated genes)
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## Level 2: 16 nodes to be scored (11236 eliminated genes)
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## Level 1: 1 nodes to be scored (12868 eliminated genes)GOTable(ResMFUp$ResSel, maxGO=20)Result visualization: Barplot
topGOBarplotAll(TopGOResAll=ResMFAll$ResSel, TopGOResDown=ResMFDown$ResSel, TopGOResUp=ResMFUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
Top Terms associated Genes
All
plotGenesInTerm_v2(ResMFAll$ResSel, ResMFAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
Down
plotGenesInTerm_v2(ResMFDown$ResSel, ResMFDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
Up
plotGenesInTerm_v2(ResMFUp$ResSel, ResMFUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
5.4 TopGO analysis: Cellular Component
On the basis of the analysis settings, the enrichment for Cellular Component IS performed.
Cellular Component Enrichment for ALL modulated genes: 3541 genes
CCann <- topGO::annFUN.org(whichOnto='CC', feasibleGenes=names(GeneVectors$DEGenes),
mapping='org.Hs.eg.db', ID='symbol') %>% inverseList()
# Wrapper function for topGO analysis
ResCCAll <- topGOResults(Genes=GeneVectors$DEGenes, gene2GO=CCann, ontology='CC',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCAll')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 620 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 13: 1 nodes to be scored (0 eliminated genes)
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## Level 12: 3 nodes to be scored (0 eliminated genes)
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## Level 11: 26 nodes to be scored (30 eliminated genes)
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## Level 10: 67 nodes to be scored (83 eliminated genes)
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## Level 9: 94 nodes to be scored (843 eliminated genes)
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## Level 8: 90 nodes to be scored (2741 eliminated genes)
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## Level 7: 91 nodes to be scored (5047 eliminated genes)
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## Level 6: 86 nodes to be scored (8289 eliminated genes)
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## Level 5: 72 nodes to be scored (9822 eliminated genes)
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## Level 4: 48 nodes to be scored (11670 eliminated genes)
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## Level 3: 39 nodes to be scored (12709 eliminated genes)
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## Level 2: 2 nodes to be scored (13074 eliminated genes)
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## Level 1: 1 nodes to be scored (13169 eliminated genes)
#write.table(ResCCAll$ResAll, file=paste0(OutputFolder, 'TopGO/CCAllResults.txt'), sep='\t', row.names=FALSE)Cellular Component Enrichment for DOWN-REGULATED genes: 1370 genes
# Wrapper function for topGO analysis
ResCCDown <- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=CCann, ontology='CC',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 576 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 12: 1 nodes to be scored (0 eliminated genes)
##
## Level 11: 24 nodes to be scored (0 eliminated genes)
##
## Level 10: 62 nodes to be scored (24 eliminated genes)
##
## Level 9: 85 nodes to be scored (817 eliminated genes)
##
## Level 8: 86 nodes to be scored (2705 eliminated genes)
##
## Level 7: 86 nodes to be scored (4988 eliminated genes)
##
## Level 6: 75 nodes to be scored (8281 eliminated genes)
##
## Level 5: 69 nodes to be scored (9804 eliminated genes)
##
## Level 4: 46 nodes to be scored (11662 eliminated genes)
##
## Level 3: 39 nodes to be scored (12709 eliminated genes)
##
## Level 2: 2 nodes to be scored (13074 eliminated genes)
##
## Level 1: 1 nodes to be scored (13169 eliminated genes)GOTable(ResCCDown$ResSel, maxGO=20)Cellular Component Analysis for UP-REGULATED genes: 2171 genes
# Wrapper function for topGO analysis
ResCCUp <- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=CCann, ontology='CC',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 539 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 13: 1 nodes to be scored (0 eliminated genes)
##
## Level 12: 3 nodes to be scored (0 eliminated genes)
##
## Level 11: 21 nodes to be scored (30 eliminated genes)
##
## Level 10: 51 nodes to be scored (83 eliminated genes)
##
## Level 9: 75 nodes to be scored (747 eliminated genes)
##
## Level 8: 76 nodes to be scored (2539 eliminated genes)
##
## Level 7: 81 nodes to be scored (4750 eliminated genes)
##
## Level 6: 81 nodes to be scored (8205 eliminated genes)
##
## Level 5: 67 nodes to be scored (9784 eliminated genes)
##
## Level 4: 43 nodes to be scored (11662 eliminated genes)
##
## Level 3: 37 nodes to be scored (12707 eliminated genes)
##
## Level 2: 2 nodes to be scored (13074 eliminated genes)
##
## Level 1: 1 nodes to be scored (13169 eliminated genes)GOTable(ResCCUp$ResSel, maxGO=20)Result visualization: Barplot
topGOBarplotAll(TopGOResAll=ResCCAll$ResSel, TopGOResDown=ResCCDown$ResSel, TopGOResUp=ResCCUp$ResSel,
terms=8, pvalTh=0.01, plotTitle=NULL)
Top Terms associated Genes
All
plotGenesInTerm_v2(ResCCAll$ResSel, ResCCAll$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle=NULL, Interactive=FALSE)
Down
plotGenesInTerm_v2(ResCCDown$ResSel, ResCCDown$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Down DEGs', Interactive=FALSE, fillCol='blue')
Up
plotGenesInTerm_v2(ResCCUp$ResSel, ResCCUp$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
6. Savings
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_0.4.2Date## [1] "Fri Jul 18 19:46:37 2025"