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I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze metabolism I did this (keep only the pathways envolved in metabolism):

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with significant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?

I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze metabolism I did this (keep only the pathways envolved in metabolism):

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with significant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?

These are the 4 genes DGE and included in my kit:

  gs_name                              gene_symbol
  <chr>                                <chr>      
1 KEGG_ARGININE_AND_PROLINE_METABOLISM ALDH2      
2 KEGG_ARGININE_AND_PROLINE_METABOLISM ARG2       
3 KEGG_ARGININE_AND_PROLINE_METABOLISM NOS1       
4 KEGG_ARGININE_AND_PROLINE_METABOLISM ODC1    

And this are all the 59.

here all the genes dge and all the genes in pathway:

 genes_dge <- c("ALDH2", "ARG2", "NOS1", "ODC1")
 all_genes <- c("ACY1", "AGMAT", "ALDH18A1", "ALDH1B1", "ALDH2", "ALDH3A2", "ALDH4A1", "ALDH7A1", "ALDH9A1", "AMD1", "AOC1", "ARG1", "ARG2", "ASL", "ASS1", "AZIN2", "CKB", "CKM", "CKMT1A", "CKMT1B",s"CKMT2", "CPS1", "DAO", "GAMT", "GATM", "GLS", "GLS2", "GLUD1", "GLUD2", "GLUD2", "GLUL", "GOT1", "GOT2", "LAP3", "MAOA", "MAOB", "NAGS", "NOS1", "NOS2", "NOS3", "OAT", "ODC1", "OTC", "P4HA1", "P4HA2", "P4HA3", "PRODH", "PRODH2", "PYCR1", "PYCR2", "PYCR3", "PYCR3", "SAT1","SAT2", "SMS", "SRM") 

I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze glicolysis I did this:

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with significant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?

I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze metabolism I did this (keep only the pathways envolved in metabolism):

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with significant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?

I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze glicolysis I did this:

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with signficant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?

I have a gene set containing a set of 1500 genes in which I did differential gene expression using limma voom pipeline. After this step I wanted to analyze a specific set of pathways the metabolism set from KEGG database.

hs_kegg_df <- msigdbr(species = "Homo sapiens") %>%
 dplyr::filter(
gs_cat == "C2", # This is to filter only to the C2 curated gene sets
gs_subcat == "CP:KEGG" # This is because we only want KEGG pathways
)

Then since I only want to analyze glicolysis I did this:

hs_kegg_df_with_gs_source <- subset(hs_kegg_df, grepl("^hsa00|^hsa01|^hsa02", gs_exact_source))

My problem is that, for example, for the pathway KEGG arginine and proline metabolism the set contains 54 genes in which my kit only covers 4, and these for 4 genes are Differentially expressed yet when doing: ( I decided to do ORA because with 1500 genes GSEA is not recommended).

em <- enricher(genes, minGSSize=1, maxGSSize = 20,  universe = names(gene_list), TERM2GENE=hs_kegg_df,pAdjustMethod = "fdr")  

This set does not appear with significant p value. I also tried passing to the enricher just that pathway in particular, the p value is 1. Because in hypergeometric test, the background genes are the same as the as DGE genes in all pathways giving this way 1-0=1.

How can I analyze just one pathway in particular given this specific conditions?