Loading package and necessary data
library(LEANR)
set.seed(123456)
# load network and CCM p-values
data(g2)
data(CCM.pvals)
data(gene.annots)Run LEAN on the precomputed differential expression p-values (computed using limma) in turn for each of the three CCM gene knock-out experiments. As we run these calculations with background distributions of size 10.000, they will take a little time depending on how many cores are available on your machine.
LEAN_results<-lapply(names(CCM.pvals),function(ccm){
run.lean(CCM.pvals[[ccm]], g2, n_reps = 10000, ncores = 3)
})
names(LEAN_results)<-names(CCM.pvals)Extract significant local subnetworks in each of the three experiments and show which local subnetworks are significant in all three knock-out experiments
# Extract significant local subnetworks
sign.genes<-lapply(LEAN_results,function(LEANres){
rownames(LEANres$restab[LEANres$restab[,'PLEAN']<=0.05,])
})
names(sign.genes)<-names(CCM.pvals)
# Show local subnetworks detected as significant in all three knock-outs
all.sign<-intersect(intersect(sign.genes[[1]],sign.genes[[2]]),sign.genes[[3]])
print(gene.annots[all.sign,])## ensembl_gene_id mgi_symbol entrezgene
## 10090.ENSMUSP00000107873 ENSMUSG00000001930 Vwf <NA>
## description
## 10090.ENSMUSP00000107873 Von Willebrand factor homolog [Source:MGI Symbol;Acc:MGI:98941]The local subnetwork around VWF is the only one significant in all three knock-outs. Now we extract the local subnetwork around VWF and create a Cytoscape-readable subnetwork file
# Extract information on the detected local subnetwork (around VWF)
vwf.ls.info<-get.ls.info(all.sign[1],LEAN_results$CCM2)
print(head(vwf.ls.info))
write.table(vwf.ls.info,'./VWF_subnetwork_info_CCM2.txt',sep='\t',quote=F,row.names=F)
# Create sif file representing the local subnetwork around VWF
write.ls.to.sif(all.sign[1],LEAN_results$CCM2,'./VWF_subnetwork.sif')After completing these steps, the local subnetwork around VWF can be inspected in Cytoscape by loading the network from file