ctlab / fgsea

Fast Gene Set Enrichment Analysis
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fgsea hangs forever for highly enriched pathways in the presence of repeated high scored genes #151

Open guidohooiveld opened 5 months ago

guidohooiveld commented 5 months ago

Hi Alex,

A (reproducible) issue ("GSEA hangs") was posted on the clusterProfiler GitHub. See: https://github.com/YuLab-SMU/clusterProfiler/issues/659#issuecomment-2027820878, and posts below that one.

Since clusterProfiler uses under the hood fgsea for gene set enrichment analysis, I checked whether the reported issue originates from the way input/output data is being processed by clusterProfiler, or from fgsea. It turns that I could reproduce the issue when directly using fgsea, hence this post.

Please note that the OP reported this issue when using R-4.2.2, but I could reproduce it also with the current versions of R (R-4.3.0 resp. R-4.3.3) and fgsea on both my Windows resp. Linux machines.

Also note that the issue occurs when minSize is set to 10; when minSize=11 is ued fgsea runs as expected...

For your convenience I have attached the 2 input files to this post as RData file (which I compressed into an ZIP archive in order to be able to upload it). See below how these objects were generated, also in case you would like to generate them yourselves.

I would appreciate if you could have a look at this to see whether this can be fixed. G

> ## load libraries
> library(clusterProfiler)
> library(fgsea)
> library(org.Hs.eg.db)
> 
> ## import input genes (human ENSEMBL) and GO-BP gene sets
> load("fgsea.input.Rdata")
> 
> ######
> ## if preferred, code to generate input
> 
> ## copy/paste list of input genes ('hgene_list') from:
> ## https://github.com/YuLab-SMU/clusterProfiler/issues/659#issuecomment-2027820878
> 
> 
> ## create GO-based gene sets; limit to BP
> ## 'ont' should either be "BP", "CC", "MF" or all
> library(GO.db)
> ont <- "BP" 
> 
> goterms <- AnnotationDbi::Ontology(GO.db::GOTERM)
> if (ont != "ALL") {goterms <- goterms[goterms == ont]}
> 
> term2gene.go <- AnnotationDbi::mapIds(org.Hs.eg.db,
+                                       keys=names(goterms),
+                                       column="ENTREZID",
+                                       keytype="GOALL",
+                                       multiVals='list')
'select()' returned 1:many mapping between keys and columns
> 
> ## end code to generate input.
> ######
> 
> ## manually convert ENSEMBL into ENTREZID using function bitr from clusterProfiler.
> ## when using the function gseGO from clusterProfiler, this is being done on the fly;
> ## see for gseGO function call: https://github.com/YuLab-SMU/clusterProfiler/issues/659#issuecomment-2027820878
> 
> ensembl.2.eg <- bitr( names(hgene_list),
+                       fromType="ENSEMBL",
+                       toType="ENTREZID",
+                       OrgDb="org.Hs.eg.db",
+                       drop = TRUE)
'select()' returned 1:many mapping between keys and columns
Warning message:
In bitr(names(hgene_list), fromType = "ENSEMBL", toType = "ENTREZID",  :
  0.05% of input gene IDs are fail to map...
> 
> 
> input.genes <- hgene_list[ensembl.2.eg$ENSEMBL]
> names(input.genes) <- ensembl.2.eg$ENTREZID
> ## perform GSEA
> ## with minSize = 11; works fine!
> 
> system.time({
+ 
+ res <- fgseaMultilevel(
+   pathways = term2gene.go,
+   stats = input.genes,
+   minSize = 11,
+   maxSize = 500,
+   eps = 0,
+   scoreType = c("std") )
+ 
+   })
   user  system elapsed 
   3.47    0.87   20.19 
Warning messages:
1: In preparePathwaysAndStats(pathways, stats, minSize, maxSize, gseaParam,  :
  There are ties in the preranked stats (2.19% of the list).
The order of those tied genes will be arbitrary, which may produce unexpected results.
2: In fgseaMultilevel(pathways = term2gene.go, stats = input.genes,  :
  There were 8 pathways for which P-values were not calculated properly due to unbalanced (positive and negative) gene-level statistic values. For such pathways pval, padj, NES, log2err are set to NA. You can try to increase the value of the argument nPermSimple (for example set it nPermSimple = 10000)
3: In fgseaMultilevel(pathways = term2gene.go, stats = input.genes,  :
  For some of the pathways the P-values were likely overestimated. For such pathways log2err is set to NA.
> 
> ## perform GSEA
> ## now with minSize = 10; run was aborted after 5 mins since it wasn't finished by then...
> 
> system.time({
+ 
+ res <- fgseaMultilevel(
+   pathways = term2gene.go,
+   stats = input.genes,
+   minSize = 10,
+   maxSize = 500,
+   eps = 0,
+   scoreType = c("std") )
+ 
+   })

Warning messages:
1: In preparePathwaysAndStats(pathways, stats, minSize, maxSize, gseaParam,  :
  There are ties in the preranked stats (2.19% of the list).
The order of those tied genes will be arbitrary, which may produce unexpected results.
2: In fgseaMultilevel(pathways = term2gene.go, stats = input.genes,  :
  There were 4 pathways for which P-values were not calculated properly due to unbalanced (positive and negative) gene-level statistic values. For such pathways pval, padj, NES, log2err are set to NA. You can try to increase the value of the argument nPermSimple (for example set it nPermSimple = 10000)

Timing stopped at: 3.07 0.91 592.6
> 
>

sessionInfo() Windows machine:

> sessionInfo()
R version 4.3.0 (2023-04-21 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 19042)

Matrix products: default

locale:
[1] LC_COLLATE=English_United States.utf8 
[2] LC_CTYPE=English_United States.utf8   
[3] LC_MONETARY=English_United States.utf8
[4] LC_NUMERIC=C                          
[5] LC_TIME=English_United States.utf8    

time zone: Europe/Amsterdam
tzcode source: internal

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods  
[8] base     

other attached packages:
[1] org.Hs.eg.db_3.18.0    AnnotationDbi_1.64.1   IRanges_2.36.0        
[4] S4Vectors_0.40.2       Biobase_2.62.0         BiocGenerics_0.48.1   
[7] fgsea_1.28.0           clusterProfiler_4.10.1

loaded via a namespace (and not attached):
 [1] DBI_1.2.2               bitops_1.0-7            shadowtext_0.1.3       
 [4] gson_0.1.0              gridExtra_2.3           rlang_1.1.3            
 [7] magrittr_2.0.3          DOSE_3.28.2             compiler_4.3.0         
[10] RSQLite_2.3.6           png_0.1-8               vctrs_0.6.5            
[13] reshape2_1.4.4          stringr_1.5.1           pkgconfig_2.0.3        
[16] crayon_1.5.2            fastmap_1.1.1           XVector_0.42.0         
[19] ggraph_2.2.1            utf8_1.2.4              HDO.db_0.99.1          
[22] enrichplot_1.23.1.992   purrr_1.0.2             bit_4.0.5              
[25] zlibbioc_1.48.2         cachem_1.0.8            aplot_0.2.2            
[28] GenomeInfoDb_1.38.8     jsonlite_1.8.8          blob_1.2.4             
[31] BiocParallel_1.36.0     tweenr_2.0.3            parallel_4.3.0         
[34] R6_2.5.1                stringi_1.8.3           RColorBrewer_1.1-3     
[37] GOSemSim_2.29.1.001     Rcpp_1.0.12             snow_0.4-4             
[40] Matrix_1.6-5            splines_4.3.0           igraph_2.0.3           
[43] tidyselect_1.2.1        qvalue_2.34.0           viridis_0.6.5          
[46] codetools_0.2-20        lattice_0.22-6          tibble_3.2.1           
[49] plyr_1.8.9              treeio_1.26.0           withr_3.0.0            
[52] KEGGREST_1.42.0         gridGraphics_0.5-1      scatterpie_0.2.1       
[55] polyclip_1.10-6         Biostrings_2.70.3       pillar_1.9.0           
[58] ggtree_3.10.1           ggfun_0.1.4             generics_0.1.3         
[61] RCurl_1.98-1.14         ggplot2_3.5.0           munsell_0.5.1          
[64] scales_1.3.0            tidytree_0.4.6          glue_1.7.0             
[67] lazyeval_0.2.2          tools_4.3.0             data.table_1.15.4      
[70] fs_1.6.3                graphlayouts_1.1.1      fastmatch_1.1-4        
[73] tidygraph_1.3.1         cowplot_1.1.3           grid_4.3.0             
[76] tidyr_1.3.1             ape_5.7-1               colorspace_2.1-0       
[79] nlme_3.1-164            GenomeInfoDbData_1.2.11 patchwork_1.2.0        
[82] ggforce_0.4.2           cli_3.6.2               fansi_1.0.6            
[85] viridisLite_0.4.2       dplyr_1.1.4             gtable_0.3.4           
[88] yulab.utils_0.1.4       digest_0.6.35           ggrepel_0.9.5          
[91] ggplotify_0.1.2         farver_2.1.1            memoise_2.0.1          
[94] lifecycle_1.0.4         httr_1.4.7              GO.db_3.18.0           
[97] bit64_4.0.5             MASS_7.3-60.0.1        
> 

sessionInfo() Linux machine:

> sessionInfo()
R version 4.3.3 (2024-02-29)
Platform: x86_64-redhat-linux-gnu (64-bit)
Running under: Fedora Linux 39 (Thirty Nine)

Matrix products: default
BLAS/LAPACK: FlexiBLAS OPENBLAS-OPENMP;  LAPACK version 3.11.0

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       

time zone: Europe/Amsterdam
tzcode source: system (glibc)

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods  
[8] base     

other attached packages:
[1] org.Hs.eg.db_3.18.0    AnnotationDbi_1.64.1   IRanges_2.36.0        
[4] S4Vectors_0.40.2       Biobase_2.62.0         BiocGenerics_0.48.1   
[7] fgsea_1.28.0           clusterProfiler_4.10.1

loaded via a namespace (and not attached):
 [1] DBI_1.2.2               bitops_1.0-7            shadowtext_0.1.3       
 [4] gson_0.1.0              gridExtra_2.3           rlang_1.1.3            
 [7] magrittr_2.0.3          DOSE_3.28.2             compiler_4.3.3         
[10] RSQLite_2.3.6           png_0.1-8               vctrs_0.6.5            
[13] reshape2_1.4.4          stringr_1.5.1           pkgconfig_2.0.3        
[16] crayon_1.5.2            fastmap_1.1.1           XVector_0.42.0         
[19] ggraph_2.2.1            utf8_1.2.4              HDO.db_0.99.1          
[22] enrichplot_1.22.0       purrr_1.0.2             bit_4.0.5              
[25] zlibbioc_1.48.2         cachem_1.0.8            aplot_0.2.2            
[28] GenomeInfoDb_1.38.8     jsonlite_1.8.8          blob_1.2.4             
[31] BiocParallel_1.36.0     tweenr_2.0.3            parallel_4.3.3         
[34] R6_2.5.1                stringi_1.8.3           RColorBrewer_1.1-3     
[37] GOSemSim_2.28.1         Rcpp_1.0.12             Matrix_1.6-5           
[40] splines_4.3.3           igraph_2.0.3            tidyselect_1.2.1       
[43] qvalue_2.34.0           viridis_0.6.5           codetools_0.2-20       
[46] lattice_0.22-6          tibble_3.2.1            plyr_1.8.9             
[49] treeio_1.26.0           withr_3.0.0             KEGGREST_1.42.0        
[52] gridGraphics_0.5-1      scatterpie_0.2.2        polyclip_1.10-6        
[55] Biostrings_2.70.3       pillar_1.9.0            ggtree_3.10.1          
[58] ggfun_0.1.4             generics_0.1.3          RCurl_1.98-1.14        
[61] ggplot2_3.5.0           munsell_0.5.1           scales_1.3.0           
[64] tidytree_0.4.6          glue_1.7.0              lazyeval_0.2.2         
[67] tools_4.3.3             data.table_1.15.4       fs_1.6.3               
[70] graphlayouts_1.1.1      fastmatch_1.1-4         tidygraph_1.3.1        
[73] cowplot_1.1.3           grid_4.3.3              tidyr_1.3.1            
[76] ape_5.7-1               colorspace_2.1-0        nlme_3.1-164           
[79] GenomeInfoDbData_1.2.11 patchwork_1.2.0         ggforce_0.4.2          
[82] cli_3.6.2               fansi_1.0.6             viridisLite_0.4.2      
[85] dplyr_1.1.4             gtable_0.3.4            yulab.utils_0.1.4      
[88] digest_0.6.35           ggrepel_0.9.5           ggplotify_0.1.2        
[91] farver_2.1.1            memoise_2.0.1           lifecycle_1.0.4        
[94] httr_1.4.7              GO.db_3.18.0            bit64_4.0.5            
[97] MASS_7.3-60.0.1        
> 

fgsea.input.zip

assaron commented 5 months ago

@guidohooiveld thanks for the report. I can reproduce the problem. I'll check later what's going on.

assaron commented 4 months ago

To keep you updated: this is turned out to be an issue of the algorithm we were generally aware of, although not in this setting. Anyway we recently developed an approach how to properly fix it. Hopefully we'll be able to integrate the proper fix into fgsea in not so distant future, but also it's not trivial, so I can't make any ETA. As a workaround for now one could add random noise to the input scores, and everything should start working fine:

res <- fgseaMultilevel(
    pathways = term2gene.go,
    stats = input.genes+rnorm(length(input.genes), sd=0.001),
    minSize = 10,
    maxSize = 500,
    eps = 0,
    scoreType = c("std") )
jinxmeng commented 3 weeks ago

Hello, I believe this post might help solve some problems (https://www.biostars.org/p/327699/). Running calculations in parallel on a Windows system can be challenging, but using the code register(SerialParam()) can force the machine to run in Serial mode. On a Linux system, I tested the code without any issues and without needing any adjustments. I'm not familiar with the underlying operating mechanism, though.

jinxmeng commented 3 weeks ago

Hello, I believe this post might help solve some problems (https://www.biostars.org/p/327699/). Running calculations in parallel on a Windows system can be challenging, but using the code register(SerialParam()) can force the machine to run in Serial mode. On a Linux system, I tested the code without any issues and without needing any adjustments. I'm not familiar with the underlying operating mechanism, though.

sessionInfo()

R version 4.3.1 (2023-06-16 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 11 x64 (build 22631)

Matrix products: default

locale:
[1] LC_COLLATE=Chinese (Simplified)_China.utf8  LC_CTYPE=Chinese (Simplified)_China.utf8   
[3] LC_MONETARY=Chinese (Simplified)_China.utf8 LC_NUMERIC=C                               
[5] LC_TIME=Chinese (Simplified)_China.utf8    

time zone: Asia/Shanghai
tzcode source: internal

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] BiocParallel_1.34.2   org.Mm.eg.db_3.17.0   AnnotationDbi_1.62.2  IRanges_2.34.1        S4Vectors_0.38.2     
 [6] Biobase_2.60.0        BiocGenerics_0.46.0   enrichplot_1.20.3     clusterProfiler_4.8.3 ggpubr_0.6.0         
[11] ggplot2_3.5.1         purrr_1.0.2           tibble_3.2.1          dplyr_1.1.4           tidyr_1.3.1          

loaded via a namespace (and not attached):
  [1] DBI_1.2.3               bitops_1.0-8            gson_0.1.0              shadowtext_0.1.4        gridExtra_2.3          
  [6] rlang_1.1.4             magrittr_2.0.3          DOSE_3.26.2             compiler_4.3.1          RSQLite_2.3.7          
 [11] png_0.1-8               vctrs_0.6.5             reshape2_1.4.4          stringr_1.5.1           pkgconfig_2.0.3        
 [16] crayon_1.5.3            fastmap_1.2.0           backports_1.5.0         XVector_0.40.0          ggraph_2.2.1           
 [21] utf8_1.2.4              HDO.db_0.99.1           bit_4.0.5               zlibbioc_1.46.0         cachem_1.1.0           
 [26] aplot_0.2.3             jsonlite_1.8.8          GenomeInfoDb_1.36.4     blob_1.2.4              tweenr_2.0.3           
 [31] broom_1.0.6             parallel_4.3.1          R6_2.5.1                stringi_1.8.4           RColorBrewer_1.1-3     
 [36] car_3.1-2               GOSemSim_2.26.1         Rcpp_1.0.13             snow_0.4-4              downloader_0.4         
 [41] pacman_0.5.1            Matrix_1.5-4.1          splines_4.3.1           igraph_2.0.3            tidyselect_1.2.1       
 [46] qvalue_2.32.0           abind_1.4-5             viridis_0.6.5           codetools_0.2-20        lattice_0.22-6         
 [51] plyr_1.8.9              treeio_1.24.3           withr_3.0.1             KEGGREST_1.40.1         gridGraphics_0.5-1     
 [56] scatterpie_0.2.4        polyclip_1.10-7         Biostrings_2.68.1       ggtree_3.8.2            pillar_1.9.0           
 [61] carData_3.0-5           ggfun_0.1.6             generics_0.1.3          RCurl_1.98-1.16         tidytree_0.4.6         
 [66] munsell_0.5.1           scales_1.3.0            glue_1.7.0              lazyeval_0.2.2          tools_4.3.1            
 [71] data.table_1.16.0       fgsea_1.26.0            ggsignif_0.6.4          fs_1.6.4                graphlayouts_1.1.1     
 [76] fastmatch_1.1-4         tidygraph_1.3.1         cowplot_1.1.3           grid_4.3.1              ape_5.8                
 [81] colorspace_2.1-1        nlme_3.1-166            patchwork_1.2.0         GenomeInfoDbData_1.2.10 ggforce_0.4.2          
 [86] cli_3.6.3               fansi_1.0.6             viridisLite_0.4.2       gtable_0.3.5            rstatix_0.7.2          
 [91] yulab.utils_0.1.7       digest_0.6.37           ggrepel_0.9.5           ggplotify_0.1.2         farver_2.1.2           
 [96] memoise_2.0.1           lifecycle_1.0.4         httr_1.4.7              GO.db_3.17.0            bit64_4.0.5            
[101] MASS_7.3-60 
yeroslaviz commented 2 weeks ago

The solution with SerialParam() worked nicely. thanks

assaron commented 2 weeks ago

@jinxmeng Thanks for your comment, but I suspect it to be a different bug. It'd be great if you could provide more background, like was there anything special about your input and how reproducible it in your settings. We don't test it on Windows machines too much, but my understanding was that it should run OK, even without SerialParam() setting.

assaron commented 2 weeks ago

@yeroslaviz Similarly, I'm not sure what's going on in your case. I saw your post on Biostars, apparently you use Mac, not Windows, and also had nproc=1 parameter. The latter, in my mind, should be effectively the same as setting SerialParam() mode, so it's weird that it helped. Could you also provide more background on your case?

jinxmeng commented 2 weeks ago

@assaron Thank you for your response. I don't fully understand the operating mechanism here. I will provide the relevant files and code for you to test. Thank you! My computer system is Microsoft Windows 11, and my CPU is 'Intel64 Family 6 Model 183 Stepping 1 GenuineIntel ~2100 MHz' (Intel(R) Core(TM) i7-14700HX, 2100 MHz, 20 cores).

> library(dplyr)
> library(clusterProfiler)
> gene_list <- readRDS("gene_list.rds")
> gseKEGG <- gseKEGG(gene_list, organism = "mmu", minGSSize = 10, maxGSSize = 1000, pvalueCutoff = 1)
preparing geneSet collections...
GSEA analysis...
#The process lasted for 1 minute without any response.

> register(SerialParam())
> system.time(gseKEGG <- gseKEGG(gene_list, organism = "mmu", minGSSize = 10, maxGSSize = 1000, pvalueCutoff = 1))
preparing geneSet collections...
GSEA analysis...
leading edge analysis...
done...
用户 系统 流逝 
0.64 0.02 1.86 
Warning messages:
1: In preparePathwaysAndStats(pathways, stats, minSize, maxSize, gseaParam,  :
  There are ties in the preranked stats (0.02% of the list).
The order of those tied genes will be arbitrary, which may produce unexpected results.
2: In preparePathwaysAndStats(pathways, stats, minSize, maxSize, gseaParam,  :
  There are duplicate gene names, fgsea may produce unexpected results.
3: In fgseaMultilevel(pathways = pathways, stats = stats, minSize = minSize,  :
  For some pathways, in reality P-values are less than 1e-10. You can set the `eps` argument to zero for better estimation.
#The process lasted for 1.86 seconds and produced the output.

> sessionInfo()
R version 4.3.3 (2024-02-29 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 11 x64 (build 22631)

Matrix products: default

locale:
[1] LC_COLLATE=Chinese (Simplified)_China.utf8  LC_CTYPE=Chinese (Simplified)_China.utf8    LC_MONETARY=Chinese (Simplified)_China.utf8
[4] LC_NUMERIC=C                                LC_TIME=Chinese (Simplified)_China.utf8    

time zone: Asia/Shanghai
tzcode source: internal

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] BiocParallel_1.36.0    org.Mm.eg.db_3.18.0    AnnotationDbi_1.64.1   IRanges_2.36.0         S4Vectors_0.40.2       Biobase_2.62.0        
 [7] BiocGenerics_0.48.1    enrichplot_1.22.0      clusterProfiler_4.12.2 cowplot_1.1.3          data.table_1.15.4      harmony_1.2.0         
[13] Rcpp_1.0.13            Seurat_5.1.0           SeuratObject_5.0.2     sp_2.1-4               stringr_1.5.1          metafor_4.6-0         
[19] numDeriv_2016.8-1.1    metadat_1.2-0          Matrix_1.6-5           ggpubr_0.6.0           ggplot2_3.5.1          purrr_1.0.2           
[25] tibble_3.2.1           dplyr_1.1.4            tidyr_1.3.1           

loaded via a namespace (and not attached):
  [1] RcppAnnoy_0.0.22        splines_4.3.3           later_1.3.2             ggplotify_0.1.2         bitops_1.0-7           
  [6] polyclip_1.10-7         fastDummies_1.7.4       httr2_1.0.2             lifecycle_1.0.4         rstatix_0.7.2          
 [11] globals_0.16.3          lattice_0.22-5          MASS_7.3-60.0.1         backports_1.5.0         magrittr_2.0.3         
 [16] limma_3.60.3            plotly_4.10.4           httpuv_1.6.15           sctransform_0.4.1       spam_2.10-0            
 [21] spatstat.sparse_3.1-0   reticulate_1.38.0       pbapply_1.7-2           DBI_1.2.3               RColorBrewer_1.1-3     
 [26] abind_1.4-5             zlibbioc_1.48.0         Rtsne_0.17              presto_1.0.0            ggraph_2.2.1           
 [31] RCurl_1.98-1.14         yulab.utils_0.1.6       tweenr_2.0.3            rappdirs_0.3.3          GenomeInfoDbData_1.2.11
 [36] ggrepel_0.9.5           irlba_2.3.5.1           listenv_0.9.1           spatstat.utils_3.0-5    tidytree_0.4.6         
 [41] goftest_1.2-3           RSpectra_0.16-2         spatstat.random_3.3-1   fitdistrplus_1.2-1      parallelly_1.38.0      
 [46] leiden_0.4.3.1          codetools_0.2-19        ggforce_0.4.2           DOSE_3.28.2             tidyselect_1.2.1       
 [51] aplot_0.2.3             farver_2.1.2            viridis_0.6.5           matrixStats_1.3.0       spatstat.explore_3.3-1 
 [56] mathjaxr_1.6-0          jsonlite_1.8.8          tidygraph_1.3.1         progressr_0.14.0        ggridges_0.5.6         
 [61] survival_3.5-8          systemfonts_1.1.0       tools_4.3.3             treeio_1.26.0           ragg_1.3.2             
 [66] snow_0.4-4              ica_1.0-3               glue_1.7.0              gridExtra_2.3           qvalue_2.34.0          
 [71] GenomeInfoDb_1.38.8     withr_3.0.1             fastmap_1.2.0           fansi_1.0.6             digest_0.6.36          
 [76] gridGraphics_0.5-1      R6_2.5.1                mime_0.12               textshaping_0.4.0       colorspace_2.1-1       
 [81] GO.db_3.18.0            scattermore_1.2         tensor_1.5              spatstat.data_3.1-2     RSQLite_2.3.6          
 [86] RhpcBLASctl_0.23-42     utf8_1.2.4              generics_0.1.3          graphlayouts_1.1.1      httr_1.4.7             
 [91] htmlwidgets_1.6.4       scatterpie_0.2.3        uwot_0.2.2              pkgconfig_2.0.3         gtable_0.3.5           
 [96] blob_1.2.4              lmtest_0.9-40           XVector_0.42.0          shadowtext_0.1.4        htmltools_0.5.8.1      
[101] carData_3.0-5           fgsea_1.30.0            dotCall64_1.1-1         scales_1.3.0            png_0.1-8              
[106] spatstat.univar_3.0-0   ggfun_0.1.5             rstudioapi_0.16.0       reshape2_1.4.4          nlme_3.1-164           
[111] zoo_1.8-12              cachem_1.1.0            KernSmooth_2.23-22      HDO.db_0.99.1           parallel_4.3.3         
[116] miniUI_0.1.1.1          pillar_1.9.0            grid_4.3.3              vctrs_0.6.5             RANN_2.6.1             
[121] promises_1.3.0          car_3.1-2               xtable_1.8-4            cluster_2.1.6           cli_3.6.2              
[126] compiler_4.3.3          rlang_1.1.3             crayon_1.5.3            future.apply_1.11.2     ggsignif_0.6.4         
[131] labeling_0.4.3          fs_1.6.4                plyr_1.8.9              stringi_1.8.4           viridisLite_0.4.2      
[136] deldir_2.0-4            munsell_0.5.1           Biostrings_2.70.1       lazyeval_0.2.2          spatstat.geom_3.3-2    
[141] GOSemSim_2.28.1         pacman_0.5.1            RcppHNSW_0.6.0          patchwork_1.2.0         bit64_4.0.5            
[146] future_1.34.0           KEGGREST_1.42.0         statmod_1.5.0           shiny_1.9.1             ROCR_1.0-11            
[151] igraph_2.0.3            broom_1.0.6             memoise_2.0.1           ggtree_3.10.1           fastmatch_1.1-4        
[156] bit_4.0.5               gson_0.1.0              ape_5.8          

gene_list.zip

yeroslaviz commented 2 weeks ago

@assaron - you're correct. the serialParam() didn't solve the problem, when I tried it again, but your suggestion to add random noise did. So, for now I'm happy about it.

That's also the reason, why I deleted the comment on Biostar.

thanks for the solution. It would be great if you can also fix the problem in the future.