cpg.annotate() in DMRcate error
1
0
Entering edit mode
Grace • 0
@609f1b8d
Last seen 19 months ago
United Kingdom

Hello, I know similar questions have been posted for the cpg.annotate() function in DMRcate() for DMR analysis with EPIC array data (no bisulphite sequencing), but I am really stuck and would massively appreciate any help!

Code should be placed in three backticks as shown below

 myannotation <- cpg.annotate("array", MSw, arraytype = "EPIC", analysis.type="differential", design=design, coef=2)

### returning error below

Error in if (nsig == 0) { : missing value where TRUE/FALSE needed
In addition: Warning messages:
1: In logit2(assay(object, "Beta")) : NaNs produced
2: In logit2(assay(object, "Beta")) : NaNs produced
3: Partial NA coefficients for 123812 probe(s) 

##the MSw object was produced with the following code and looks as such:
MSw <- getM(MsetSw, type = "beta", betaThreshold = 0.001)
![MSw table][1]
sessionInfo( )
R version 4.2.2 (2022-10-31 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 19044)

Matrix products: default

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

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

other attached packages:
 [1] DMRcate_2.12.0                                     
 [2] circlize_0.4.15                                    
 [3] reshape2_1.4.4                                     
 [4] plyr_1.8.8                                         
 [5] corpcor_1.6.10                                     
 [6] CpGassoc_2.60                                      
 [7] data.table_1.14.8                                  
 [8] qqman_0.1.8                                        
 [9] tidyr_1.3.0                                        
[10] pvclust_2.2-0                                      
[11] sqldf_0.4-11                                       
[12] RSQLite_2.3.1                                      
[13] gsubfn_0.7                                         
[14] proto_1.0.0                                        
[15] pcaMethods_1.90.0                                  
[16] sva_3.46.0                                         
[17] BiocParallel_1.32.6                                
[18] genefilter_1.80.3                                  
[19] mgcv_1.8-42                                        
[20] nlme_3.1-162                                       
[21] dplyr_1.1.1                                        
[22] limma_3.54.2                                       
[23] WGCNA_1.72-1                                       
[24] fastcluster_1.2.3                                  
[25] dynamicTreeCut_1.63-1                              
[26] GO.db_3.16.0                                       
[27] AnnotationDbi_1.60.2                               
[28] missMethyl_1.32.1                                  
[29] IlluminaHumanMethylationEPICanno.ilm10b4.hg19_0.6.0
[30] MatrixEQTL_2.3                                     
[31] IlluminaHumanMethylation450kanno.ilmn12.hg19_0.6.1 
[32] IlluminaHumanMethylation450kmanifest_0.4.0         
[33] FlowSorted.Blood.EPIC_2.2.0                        
[34] ExperimentHub_2.6.0                                
[35] AnnotationHub_3.6.0                                
[36] BiocFileCache_2.6.1                                
[37] dbplyr_2.3.2                                       
[38] FlowSorted.Blood.450k_1.36.0                       
[39] IlluminaHumanMethylationEPICanno.ilm10b2.hg19_0.6.0
[40] IlluminaHumanMethylationEPICmanifest_0.3.0         
[41] minfi_1.44.0                                       
[42] bumphunter_1.40.0                                  
[43] locfit_1.5-9.7                                     
[44] iterators_1.0.14                                   
[45] foreach_1.5.2                                      
[46] Biostrings_2.66.0                                  
[47] XVector_0.38.0                                     
[48] SummarizedExperiment_1.28.0                        
[49] Biobase_2.58.0                                     
[50] MatrixGenerics_1.10.0                              
[51] matrixStats_0.63.0                                 
[52] GenomicRanges_1.50.2                               
[53] GenomeInfoDb_1.34.9                                
[54] IRanges_2.32.0                                     
[55] S4Vectors_0.36.2                                   
[56] BiocGenerics_0.44.0                                

loaded via a namespace (and not attached):
  [1] rappdirs_0.3.3                rtracklayer_1.58.0           
  [3] R.methodsS3_1.8.2             ggplot2_3.4.2                
  [5] bit64_4.0.5                   knitr_1.42                   
  [7] DelayedArray_0.23.2           R.utils_2.12.2               
  [9] rpart_4.1.19                  KEGGREST_1.38.0              
 [11] RCurl_1.98-1.12               GEOquery_2.66.0              
 [13] AnnotationFilter_1.22.0       doParallel_1.0.17            
 [15] generics_0.1.3                GenomicFeatures_1.50.4       
 [17] preprocessCore_1.60.2         chron_2.3-60                 
 [19] bit_4.0.5                     tzdb_0.3.0                   
 [21] xml2_1.3.3                    httpuv_1.6.9                 
 [23] xfun_0.38                     hms_1.1.3                    
 [25] evaluate_0.20                 promises_1.2.0.1             
 [27] fansi_1.0.4                   restfulr_0.0.15              
 [29] scrime_1.3.5                  progress_1.2.2               
 [31] DBI_1.1.3                     htmlwidgets_1.6.2            
 [33] reshape_0.8.9                 purrr_1.0.1                  
 [35] ellipsis_0.3.2                backports_1.4.1              
 [37] permute_0.9-7                 calibrate_1.7.7              
 [39] annotate_1.76.0               biomaRt_2.54.1               
 [41] deldir_1.0-6                  sparseMatrixStats_1.10.0     
 [43] vctrs_0.6.2                   ensembldb_2.22.0             
 [45] cachem_1.0.7                  Gviz_1.42.1                  
 [47] BSgenome_1.66.3               checkmate_2.1.0              
 [49] GenomicAlignments_1.34.1      prettyunits_1.1.1            
 [51] mclust_6.0.0                  cluster_2.1.4                
 [53] lazyeval_0.2.2                crayon_1.5.2                 
 [55] edgeR_3.40.2                  pkgconfig_2.0.3              
 [57] ProtGenerics_1.30.0           nnet_7.3-18                  
 [59] rlang_1.1.0                   lifecycle_1.0.3              
 [61] filelock_1.0.2                dichromat_2.0-0.1            
 [63] tcltk_4.2.2                   rngtools_1.5.2               
 [65] base64_2.0.1                  Matrix_1.5-4                 
 [67] Rhdf5lib_1.20.0               base64enc_0.1-3              
 [69] GlobalOptions_0.1.2           png_0.1-8                    
 [71] rjson_0.2.21                  bitops_1.0-7                 
 [73] R.oo_1.25.0                   rhdf5filters_1.10.1          
 [75] blob_1.2.4                    DelayedMatrixStats_1.20.0    
 [77] doRNG_1.8.6                   shape_1.4.6                  
 [79] stringr_1.5.0                 nor1mix_1.3-0                
 [81] readr_2.1.4                   jpeg_0.1-10                  
 [83] scales_1.2.1                  memoise_2.0.1                
 [85] magrittr_2.0.3                zlibbioc_1.44.0              
 [87] compiler_4.2.2                BiocIO_1.8.0                 
 [89] RColorBrewer_1.1-3            illuminaio_0.40.0            
 [91] DSS_2.46.0                    Rsamtools_2.14.0             
 [93] cli_3.6.1                     htmlTable_2.4.1              
 [95] Formula_1.2-5                 MASS_7.3-58.3                
 [97] tidyselect_1.2.0              stringi_1.7.12               
 [99] yaml_2.3.7                    askpass_1.1                  
[101] latticeExtra_0.6-30           grid_4.2.2                   
[103] VariantAnnotation_1.44.1      tools_4.2.2                  
[105] rstudioapi_0.14               foreign_0.8-84               
[107] bsseq_1.34.0                  gridExtra_2.3                
[109] digest_0.6.31                 BiocManager_1.30.20          
[111] shiny_1.7.4                   quadprog_1.5-8               
[113] Rcpp_1.0.10                   siggenes_1.72.0              
[115] BiocVersion_3.16.0            later_1.3.0                  
[117] org.Hs.eg.db_3.16.0           httr_1.4.5                   
[119] biovizBase_1.46.0             colorspace_2.1-0             
[121] XML_3.99-0.14                 splines_4.2.2                
[123] statmod_1.5.0                 multtest_2.54.0              
[125] xtable_1.8-4                  R6_2.5.1                     
[127] Hmisc_5.0-1                   pillar_1.9.0                 
[129] htmltools_0.5.5               mime_0.12                    
[131] glue_1.6.2                    fastmap_1.1.1                
[133] interactiveDisplayBase_1.36.0 beanplot_1.3.1               
[135] codetools_0.2-19              utf8_1.2.3                   
[137] lattice_0.21-8                tibble_3.2.1                 
[139] curl_5.0.0                    gtools_3.9.4                 
[141] openssl_2.0.6                 interp_1.1-4                 
[143] survival_3.5-5                rmarkdown_2.21               
[145] munsell_0.5.0                 rhdf5_2.42.1                 
[147] GenomeInfoDbData_1.2.9        HDF5Array_1.26.0             
[149] impute_1.72.3                 gtable_0.3.3                 
>
DNAMethylation methylationArrayAnalysis DMRcate MethylationArrayData MethylationArray • 1.5k views
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0
Entering edit mode

I also get the same error message if I specify what=c("Beta") as an argument :)

myannotation <- cpg.annotate("array", MSw, what=c("Beta"), arraytype = "EPIC", analysis.type="differential", design=design, coef=2)

Error in if (nsig == 0) { : missing value where TRUE/FALSE needed In addition: Warning messages: 1: In logit2(assay(object, "Beta")) : NaNs produced 2: In logit2(assay(object, "Beta")) : NaNs produced 3: Partial NA coefficients for 123812 probe(s)

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0
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enter image description here

I think it could be as the table above, View(MSw), shows that many of my beta values are greater than 1. I am unsure how to fix this.

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1
Entering edit mode
Tim Peters ▴ 200
@tim-peters-7579
Last seen 3 days ago
Australia

Hi Grace,

Looks like these are M-values instead of beta values. Try:

myannotation <- cpg.annotate("array", MSw, what="M", arraytype = "EPIC", analysis.type="differential", design=design, coef=2)

Cheers, Tim

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Thanks Tim, I will try this now, I am very grateful.

Just to clarify though, if you look at my code for making the MSw, it specifies argument for 'type = beta'. I was wondering your opinion on why I have generated M-values then (attached below in the middle of the script on the right)

Many thanks, Grace

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1
Entering edit mode

Hi Grace,

When the object you pass to cpg.annotate() is a matrix, you must specify if they are M values or beta values using the what argument. In your case, the appropriate argument is what="M". The function will then take care of the rest.

I understand that the argument description for object is potentially misleading; tbh it should read "M-values or beta values" instead of just "M-values" - thank you and I'll update this.

I don't understand why you passed type="beta" to getM() though, this is unnecessary.

Cheers, Tim

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0
Entering edit mode

enter image description here

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