TopGO - CTL04
Date: July 18, 2025
Condition_1 <- params$Condition_1
Condition_2 <- params$Condition_2Top GO Analysis for Ret Agonist
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), 2969 genes are selected: 1861 up-regulated genes and 1108 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 2969 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: 2969 genes
- down-regulated genes: 1108 genes of interest
- up-regulated genes: 1861 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$TopGO))!=0, TRUE, FALSE)5.2 TopGO analysis: Biological Process
On the basis of the analysis settings, the enrichment for Biological Process IS performed.
Biological Process Analysis for ALL modulated genes: 2969 genes
BPann <- topGO::annFUN.org(whichOnto="BP", feasibleGenes=names(GeneVectors$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 --
##
## the algorithm is scoring 4774 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: 92 nodes to be scored (533 eliminated genes)
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## Level 12: 158 nodes to be scored (1434 eliminated genes)
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## Level 11: 326 nodes to be scored (3353 eliminated genes)
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## Level 10: 527 nodes to be scored (5143 eliminated genes)
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## Level 9: 678 nodes to be scored (6721 eliminated genes)
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## Level 8: 731 nodes to be scored (8583 eliminated genes)
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## Level 7: 771 nodes to be scored (10200 eliminated genes)
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## Level 6: 651 nodes to be scored (11189 eliminated genes)
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## Level 5: 404 nodes to be scored (11853 eliminated genes)
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## Level 4: 226 nodes to be scored (12259 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: 1108 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 4420 nontrivial nodes
## parameters:
## test statistic: fisher
##
## 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: 23 nodes to be scored (73 eliminated genes)
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## Level 14: 51 nodes to be scored (214 eliminated genes)
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## Level 13: 81 nodes to be scored (509 eliminated genes)
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## Level 12: 142 nodes to be scored (1373 eliminated genes)
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## Level 11: 296 nodes to be scored (3207 eliminated genes)
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## Level 10: 472 nodes to be scored (5017 eliminated genes)
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## Level 9: 613 nodes to be scored (6519 eliminated genes)
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## Level 8: 672 nodes to be scored (8368 eliminated genes)
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## Level 7: 723 nodes to be scored (10015 eliminated genes)
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## Level 6: 616 nodes to be scored (11123 eliminated genes)
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## Level 5: 387 nodes to be scored (11810 eliminated genes)
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## Level 4: 218 nodes to be scored (12251 eliminated genes)
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## Level 3: 92 nodes to be scored (12431 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$ResSel, maxGO=20)Biological Process Analysis for UP-REGULATED genes: 1861 genes
ResBPUp <- topGOResults(Genes=GeneVectors$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 4556 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: 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: 23 nodes to be scored (69 eliminated genes)
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## Level 14: 50 nodes to be scored (215 eliminated genes)
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## Level 13: 86 nodes to be scored (471 eliminated genes)
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## Level 12: 141 nodes to be scored (1333 eliminated genes)
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## Level 11: 296 nodes to be scored (3298 eliminated genes)
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## Level 10: 479 nodes to be scored (5082 eliminated genes)
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## Level 9: 639 nodes to be scored (6605 eliminated genes)
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## Level 8: 699 nodes to be scored (8438 eliminated genes)
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## Level 7: 758 nodes to be scored (10086 eliminated genes)
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## Level 6: 639 nodes to be scored (11159 eliminated genes)
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## Level 5: 400 nodes to be scored (11846 eliminated genes)
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## Level 4: 221 nodes to be scored (12259 eliminated genes)
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## Level 3: 91 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: 2969 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 805 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: 30 nodes to be scored (152 eliminated genes)
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## Level 8: 59 nodes to be scored (1213 eliminated genes)
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## Level 7: 106 nodes to be scored (3168 eliminated genes)
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## Level 6: 155 nodes to be scored (3797 eliminated genes)
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## Level 5: 183 nodes to be scored (5311 eliminated genes)
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## Level 4: 176 nodes to be scored (8260 eliminated genes)
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## Level 3: 58 nodes to be scored (10476 eliminated genes)
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## Level 2: 17 nodes to be scored (11241 eliminated genes)
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## Level 1: 1 nodes to be scored (12883 eliminated genes)Molecular Function Enrichment for DOWN-REGULATED genes: 1108 genes
ResMFDown <- topGOResults(Genes=GeneVectors$DEGenesDown, gene2GO=MFann, ontology='MF',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4060 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4530 GO terms and 5903 relations. )
##
## Annotating nodes ...............
## ( 12999 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 705 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 6 nodes to be scored (0 eliminated genes)
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## Level 10: 10 nodes to be scored (0 eliminated genes)
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## Level 9: 22 nodes to be scored (146 eliminated genes)
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## Level 8: 48 nodes to be scored (1165 eliminated genes)
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## Level 7: 93 nodes to be scored (3098 eliminated genes)
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## Level 6: 130 nodes to be scored (3710 eliminated genes)
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## Level 5: 163 nodes to be scored (5153 eliminated genes)
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## Level 4: 161 nodes to be scored (8050 eliminated genes)
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## Level 3: 54 nodes to be scored (10418 eliminated genes)
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## Level 2: 17 nodes to be scored (11210 eliminated genes)
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## Level 1: 1 nodes to be scored (12881 eliminated genes)GOTable(ResMFDown$ResSel, maxGO=20)Molecular Function Analysis for UP-REGULATED genes: 1861 genes
ResMFUp <- topGOResults(Genes=GeneVectors$DEGenesUp, gene2GO=MFann, ontology='MF',
desc=NULL, nodeSize=15, algorithm='weight01', statistic='fisher',
EnTh=params$GoEnTh, PvalTh=params$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='MFUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 4060 GO terms found. )
##
## Build GO DAG topology ..........
## ( 4530 GO terms and 5903 relations. )
##
## Annotating nodes ...............
## ( 12999 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 757 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 11: 6 nodes to be scored (0 eliminated genes)
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## Level 10: 12 nodes to be scored (0 eliminated genes)
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## Level 9: 29 nodes to be scored (137 eliminated genes)
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## Level 8: 56 nodes to be scored (1187 eliminated genes)
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## Level 7: 91 nodes to be scored (3141 eliminated genes)
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## Level 6: 145 nodes to be scored (3747 eliminated genes)
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## Level 5: 177 nodes to be scored (5098 eliminated genes)
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## Level 4: 171 nodes to be scored (8190 eliminated genes)
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## Level 3: 53 nodes to be scored (10443 eliminated genes)
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## Level 2: 16 nodes to be scored (11223 eliminated genes)
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## Level 1: 1 nodes to be scored (12868 eliminated genes)GOTable(ResMFUp$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$GOdata, SE_DEA, nterms=8, ngenes=12, plotTitle='Genes in Term - Up DEGs', Interactive=FALSE, fillCol='red')
5.4 TopGO analysis: Cellular Component
On the basis of the analysis settings, the enrichment for Cellular Component IS performed.
Cellular Component Enrichment for ALL modulated genes: 2969 genes
CCann <- topGO::annFUN.org(whichOnto='CC', feasibleGenes=names(GeneVectors$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$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCAll')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 605 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 13: 1 nodes to be scored (0 eliminated genes)
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## Level 12: 3 nodes to be scored (0 eliminated genes)
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## Level 11: 24 nodes to be scored (30 eliminated genes)
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## Level 10: 65 nodes to be scored (83 eliminated genes)
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## Level 9: 92 nodes to be scored (816 eliminated genes)
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## Level 8: 85 nodes to be scored (2665 eliminated genes)
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## Level 7: 91 nodes to be scored (4994 eliminated genes)
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## Level 6: 83 nodes to be scored (8253 eliminated genes)
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## Level 5: 71 nodes to be scored (9822 eliminated genes)
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## Level 4: 48 nodes to be scored (11662 eliminated genes)
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## Level 3: 39 nodes to be scored (12707 eliminated genes)
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## Level 2: 2 nodes to be scored (13074 eliminated genes)
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## Level 1: 1 nodes to be scored (13169 eliminated genes)
#write.table(ResCCAll$ResAll, file=paste0(OutputFolder, 'TopGO/CCAllResults.txt'), sep='\t', row.names=FALSE)Cellular Component Enrichment for DOWN-REGULATED genes: 1108 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$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCDown')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 548 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 13: 1 nodes to be scored (0 eliminated genes)
##
## Level 12: 2 nodes to be scored (0 eliminated genes)
##
## Level 11: 19 nodes to be scored (30 eliminated genes)
##
## Level 10: 57 nodes to be scored (66 eliminated genes)
##
## Level 9: 82 nodes to be scored (748 eliminated genes)
##
## Level 8: 77 nodes to be scored (2487 eliminated genes)
##
## Level 7: 80 nodes to be scored (4818 eliminated genes)
##
## Level 6: 73 nodes to be scored (8195 eliminated genes)
##
## Level 5: 68 nodes to be scored (9802 eliminated genes)
##
## Level 4: 47 nodes to be scored (11650 eliminated genes)
##
## Level 3: 39 nodes to be scored (12707 eliminated genes)
##
## Level 2: 2 nodes to be scored (13074 eliminated genes)
##
## Level 1: 1 nodes to be scored (13169 eliminated genes)GOTable(ResCCDown$ResSel, maxGO=20)Cellular Component Analysis for UP-REGULATED genes: 1861 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$GoPvalTh, minTerms=12, geneTh=4,
saveRes=TRUE, outDir=OutputFolder, fileName='CCUp')
## Gene vector contains levels: 0,1
##
## Building most specific GOs .....
## ( 1732 GO terms found. )
##
## Build GO DAG topology ..........
## ( 1926 GO terms and 3253 relations. )
##
## Annotating nodes ...............
## ( 13224 genes annotated to the GO terms. )
##
## -- Weight01 Algorithm --
##
## the algorithm is scoring 539 nontrivial nodes
## parameters:
## test statistic: fisher
##
## Level 13: 1 nodes to be scored (0 eliminated genes)
##
## Level 12: 2 nodes to be scored (0 eliminated genes)
##
## Level 11: 20 nodes to be scored (30 eliminated genes)
##
## Level 10: 49 nodes to be scored (59 eliminated genes)
##
## Level 9: 74 nodes to be scored (695 eliminated genes)
##
## Level 8: 78 nodes to be scored (2502 eliminated genes)
##
## Level 7: 83 nodes to be scored (4724 eliminated genes)
##
## Level 6: 82 nodes to be scored (8203 eliminated genes)
##
## Level 5: 66 nodes to be scored (9796 eliminated genes)
##
## Level 4: 44 nodes to be scored (11662 eliminated genes)
##
## Level 3: 37 nodes to be scored (12707 eliminated genes)
##
## Level 2: 2 nodes to be scored (13074 eliminated genes)
##
## Level 1: 1 nodes to be scored (13169 eliminated genes)GOTable(ResCCUp$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 19:36:43 2025"