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Overview

polylinkR is an R package for gene-based pathway enrichment that accounts for linkage disequilibrium among loci. This vignette demonstrates the basic workflow using the included tiny_polylinkR example dataset.

The Four-Step Workflow

The polylinkR workflow follows four sequential steps:

  1. plR_read() - Read and validate input files
  2. plR_permute() - Generate permutation nulls and compute p-values
  3. plR_rescale() - Rescale for genetic autocorrelation
  4. plR_prune() - Prune gene sets and compute FDR

Step 1: Read Input Data

First, we read the example data included with the package:

# Locate the example data
data_path <- system.file("extdata", "tiny_polylinkR", package = "polylinkR")
data_path
#> [1] "/home/runner/work/_temp/Library/polylinkR/extdata/tiny_polylinkR"

# Read the data
plr_obj <- plR_read(
  input.path = data_path,
  verbose = FALSE
)

# Check the result
class(plr_obj)
#> [1] "plR"
print(plr_obj)
#> plR class object -- output from plR_read and valid input for plR_permute 
#> NOTE:
#> Standard covariate columns not detected in obj.info
#> ** gene score deconfounding requires valid 'wt.mat' object (see ?polylinkR::plr_permute)
#> ┌────────────────────────────────────────────────────────────────────────────────┐
#>                                     set.info                                    
#> └────────────────────────────────────────────────────────────────────────────────┘
#>    setID    setName  setN
#>    <int>     <char> <int>
#> 1:     1 test_set_1     2
#> 2:     2 test_set_2     2
#> ┌────────────────────────────────────────────────────────────────────────────────┐
#>                                     obj.info                                    
#> └────────────────────────────────────────────────────────────────────────────────┘
#>    objID objStat objName    chr startpos endpos startpos.base endpos.base
#>    <int>   <num>  <char> <char>    <int>  <int>         <int>       <int>
#> 1:     1     0.5   gene1   chr1     1000   2000          1000        2000
#> 2:     2     0.6   gene2   chr1     3000   4000          3000        4000
#> 3:     3     0.7   gene3   chr1     5000   6000          5000        6000
#>    midpos
#>     <num>
#> 1:   1500
#> 2:   3500
#> 3:   5500
#> ┌────────────────────────────────────────────────────────────────────────────────┐
#>                                     set.obj                                     
#> └────────────────────────────────────────────────────────────────────────────────┘
#>    setID objID
#>    <int> <int>
#> 1:     1     1
#> 2:     1     2
#> 3:     2     2
#> 4:     2     3

The plR_read() function validates the input files and creates a plR class object containing:

  • set.info: Gene set information
  • obj.info: Gene (object) information with scores
  • set.obj: Gene-to-set mappings

Inspecting the plR Object

You can inspect the plR object using standard methods:

# Summary of the object
summary(plr_obj)
#> No tests performed. Nothing to summarise
#> 

# Structure of the object
str(plr_obj)
#> List of 3
#>  $ set.info:Classes 'data.table' and 'data.frame':   2 obs. of  3 variables:
#>   ..$ setID  : int [1:2] 1 2
#>   ..$ setName: chr [1:2] "test_set_1" "test_set_2"
#>   ..$ setN   : int [1:2] 2 2
#>   ..- attr(*, ".internal.selfref")=<externalptr> 
#>  $ obj.info:Classes 'data.table' and 'data.frame':   3 obs. of  9 variables:
#>   ..$ objID        : int [1:3] 1 2 3
#>   ..$ objStat      : num [1:3] 0.5 0.6 0.7
#>   ..$ objName      : chr [1:3] "gene1" "gene2" "gene3"
#>   ..$ chr          : chr [1:3] "chr1" "chr1" "chr1"
#>   ..$ startpos     : int [1:3] 1000 3000 5000
#>   ..$ endpos       : int [1:3] 2000 4000 6000
#>   ..$ startpos.base: int [1:3] 1000 3000 5000
#>   ..$ endpos.base  : int [1:3] 2000 4000 6000
#>   ..$ midpos       : num [1:3] 1500 3500 5500
#>   ..- attr(*, ".internal.selfref")=<externalptr> 
#>  $ set.obj :Classes 'data.table' and 'data.frame':   4 obs. of  2 variables:
#>   ..$ setID: int [1:4] 1 1 2 2
#>   ..$ objID: int [1:4] 1 2 2 3
#>   ..- attr(*, ".internal.selfref")=<externalptr> 
#>  - attr(*, "plR.data")=List of 1
#>   ..$ read.data:List of 12
#>   .. ..$ n.genes    : int 3
#>   .. ..$ n.sets     : int 2
#>   .. ..$ n.chr      : int 1
#>   .. ..$ n.cov      : num 0
#>   .. ..$ n.set.genes: int 3
#>   .. ..$ file.paths : Named chr [1:3] "/home/runner/work/_temp/Library/polylinkR/extdata/tiny_polylinkR/ObjInfo.txt" "/home/runner/work/_temp/Library/polylinkR/extdata/tiny_polylinkR/SetInfo.txt" "/home/runner/work/_temp/Library/polylinkR/extdata/tiny_polylinkR/SetObj.txt"
#>   .. .. ..- attr(*, "names")= chr [1:3] "obj.info" "set.info" "set.obj"
#>   .. ..$ coord      : chr "bp"
#>   .. ..$ get.objStat: logi FALSE
#>   .. ..$ cov.names  : NULL
#>   .. ..$ cov.info   : logi FALSE
#>   .. ..$ no.share   : logi FALSE
#>   .. ..$ pos.info   : logi TRUE
#>  - attr(*, "plR.args")=List of 1
#>   ..$ read.args:List of 20
#>   .. ..$ input.path   : chr "/home/runner/work/_temp/Library/polylinkR/extdata/tiny_polylinkR"
#>   .. ..$ obj.info.path: NULL
#>   .. ..$ set.info.path: NULL
#>   .. ..$ set.obj.path : NULL
#>   .. ..$ var.info.path: NULL
#>   .. ..$ rec.rate.path: NULL
#>   .. ..$ min.set.n    : int 2
#>   .. ..$ max.set.n    : num Inf
#>   .. ..$ group        : NULL
#>   .. ..$ map.fun      : chr "kosambi"
#>   .. ..$ obj.buffer   : num 0
#>   .. ..$ obj.stat.fun : chr "non.param"
#>   .. ..$ bin.size     : int 250
#>   .. ..$ obj.in       : NULL
#>   .. ..$ obj.out      : NULL
#>   .. ..$ set.in       : NULL
#>   .. ..$ set.out      : NULL
#>   .. ..$ set.merge    : num 0.95
#>   .. ..$ rem.genes    : logi FALSE
#>   .. ..$ verbose      : logi FALSE
#>  - attr(*, "plR.summary")=List of 1
#>   ..$ read.summary:List of 4
#>   .. ..$ removed.ID    : NULL
#>   .. ..$ repeat.ID     : NULL
#>   .. ..$ merged.ID     : NULL
#>   .. ..$ obj.stat.param: NULL
#>  - attr(*, "plR.session")=List of 1
#>   ..$ read.session:List of 2
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#>   .. .. .. ..$ platform      : chr "x86_64-pc-linux-gnu"
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Input File Formats

polylinkR requires three input files:

1. ObjInfo.txt (Gene Information)

Contains gene-level information:

  • objID: Unique gene identifier
  • objStat: Gene-level statistic (e.g., p-value, z-score)
  • Optional columns: chr, startpos, endpos (for coordinate-based analysis)

2. SetInfo.txt (Gene Set Information)

Contains gene set definitions:

  • setID: Unique set identifier
  • setName: Human-readable set name
  • setSource: Source database (optional)

3. SetObj.txt (Gene-Set Mappings)

Contains gene-to-set relationships:

  • setID: Set identifier (must match SetInfo.txt)
  • objID: Gene identifier (must match ObjInfo.txt)

Important Notes

Active Development

This package is under active development. Results should be independently validated before use in production or clinical decisions.

Algorithm Constraints

The core statistical algorithms (permutation nulls, GPD fitting, rescaling, pruning) are frozen and should not be modified without explicit maintainer approval.

Next Steps

After reading the data, you would typically proceed through the pipeline:

  1. plR_permute() - Generate permutation-based null distributions
  2. plR_rescale() - Account for genetic autocorrelation
  3. plR_prune() - Identify significant gene sets

See the function documentation (?plR_permute, ?plR_rescale, ?plR_prune) for details on subsequent steps.

Session Information

sessionInfo()
#> R version 4.5.3 (2026-03-11)
#> Platform: x86_64-pc-linux-gnu
#> Running under: Ubuntu 24.04.4 LTS
#> 
#> Matrix products: default
#> BLAS:   /usr/lib/x86_64-linux-gnu/openblas-pthread/libblas.so.3 
#> LAPACK: /usr/lib/x86_64-linux-gnu/openblas-pthread/libopenblasp-r0.3.26.so;  LAPACK version 3.12.0
#> 
#> locale:
#>  [1] LC_CTYPE=C.UTF-8       LC_NUMERIC=C           LC_TIME=C.UTF-8       
#>  [4] LC_COLLATE=C.UTF-8     LC_MONETARY=C.UTF-8    LC_MESSAGES=C.UTF-8   
#>  [7] LC_PAPER=C.UTF-8       LC_NAME=C              LC_ADDRESS=C          
#> [10] LC_TELEPHONE=C         LC_MEASUREMENT=C.UTF-8 LC_IDENTIFICATION=C   
#> 
#> time zone: UTC
#> tzcode source: system (glibc)
#> 
#> attached base packages:
#> [1] stats     graphics  grDevices utils     datasets  methods   base     
#> 
#> other attached packages:
#> [1] polylinkR_0.5.0
#> 
#> loaded via a namespace (and not attached):
#>  [1] dqrng_0.4.1           sass_0.4.10           future_1.70.0        
#>  [4] robustbase_0.99-7     distances_0.1.13      lattice_0.22-9       
#>  [7] listenv_0.10.1        digest_0.6.39         magrittr_2.0.5       
#> [10] evaluate_1.0.5        grid_4.5.3            iterators_1.0.14     
#> [13] fastmap_1.2.0         Matrix_1.7-4          foreach_1.5.2        
#> [16] jsonlite_2.0.0        mgcv_1.9-4            codetools_0.2-20     
#> [19] textshaping_1.0.5     jquerylib_0.1.4       cli_3.6.6            
#> [22] rlang_1.2.0           zigg_0.0.2            parallelly_1.46.1    
#> [25] future.apply_1.20.2   splines_4.5.3         intervals_0.15.5     
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#> [37] R6_2.6.1              zoo_1.8-15            lifecycle_1.0.5      
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#> [43] desc_1.4.3            progressr_0.19.0      pkgdown_2.2.0        
#> [46] RcppParallel_5.1.11-2 bslib_0.10.0          data.table_1.18.2.1  
#> [49] Rcpp_1.1.1            gstat_2.1-6           systemfonts_1.3.2    
#> [52] DEoptimR_1.1-4        xfun_0.57             knitr_1.51           
#> [55] nlme_3.1-168          htmltools_0.5.9       igraph_2.2.3         
#> [58] rmarkdown_2.31        xts_0.14.2            ismev_1.43           
#> [61] compiler_4.5.3        sp_2.2-1