Metafusion/Fusion Annotation

.gitignore

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+work/*
+runs/*
+logs/*
+*log
+.nextflow.log*
+.nextflow*
+results/*

bin/Metafusion_forte.sh

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+#!/bin/bash
+#STEPS
+
+# __author__      = "Alexandria Dymun"
+# __email__       = "[email protected]"
+# __contributor__ = "Anne Marie Noronha ([email protected])"
+# __version__     = "0.0.1"
+# __status__      = "Dev"
+
+
+output_ANC_RT_SG=1
+RT_call_filter=1
+blck_filter=1
+ANC_filter=1
+usage() {
+    echo "Usage: Metafusion_forte.sh --num_tools=<minNumToolsCalled> --genome_fasta <FASTA adds SEQ to fusion>  --recurrent_bedpe <blacklistFusions>  --outdir <outputDirectory> --cff <cffFile> --gene_bed <geneBedFile> --gene_info <geneInfoFile>" 1>&2;
+    exit 1;
+}
+
+# Loop through arguments and process them
+while test $# -gt 0;do
+    case $1 in
+        -n=*|--num_tools=*)
+        num_tools="${1#*=}"
+        shift
+        ;;
+        --outdir)
+        outdir="$2"
+        shift 2
+        ;;
+        --cff)
+        cff="$2"
+        shift 2
+        ;;
+        --gene_bed)
+        gene_bed="$2"
+        shift 2
+        ;;
+        --gene_info)
+        gene_info="$2"
+        shift 2
+        ;;
+        --genome_fasta)
+        genome_fasta="$2"
+        shift 2
+        ;;
+        --recurrent_bedpe)
+        recurrent_bedpe="$2"
+        shift 2
+        ;;
+        *)
+        #OTHER_ARGUMENTS+=("$1")
+        shift # Remove generic argument from processing
+        ;;
+    esac
+done
+
+if [[ ! $cff || ! $gene_info  || ! $gene_bed ]]; then
+    echo "Missing required argument"
+    usage
+fi
+
+
+mkdir $outdir
+
+#Check CFF file format:
+#Remove entries with nonconformming chromosome name
+
+all_gene_bed_chrs=`awk -F '\t' '{print $1}' $gene_bed | sort | uniq | sed 's/chr//g '`
+awk -F " " -v arr="${all_gene_bed_chrs[*]}" 'BEGIN{OFS = "\t"; split(arr,arr1); for(i in arr1) dict[arr1[i]]=""} $1 in dict && $4 in dict' $cff >  $outdir/$(basename $cff).cleaned_chr
+grep -v -f $outdir/$(basename $cff).cleaned_chr $cff > problematic_chromosomes.cff
+cff=$outdir/$(basename $cff).cleaned_chr
+
+#Rename cff
+echo Rename cff
+rename_cff_file_genes.MetaFusion.py $cff $gene_info > $outdir/$(basename $cff).renamed
+cff=$outdir/$(basename $cff).renamed
+
+#Annotate cff
+if [ $genome_fasta ]; then
+    echo Annotate cff, extract sequence surrounding breakpoint
+    reann_cff_fusion.py --cff $cff --gene_bed $gene_bed --ref_fa $genome_fasta > $outdir/$(basename $cff).reann.WITH_SEQ
+else
+    echo Annotate cff, no extraction of sequence surrounding breakpoint
+    reann_cff_fusion.py --cff $cff --gene_bed $gene_bed > $outdir/$(basename $cff).reann.NO_SEQ
+fi
+
+# Assign .cff based on SEQ or NOSEQ
+if [ $genome_fasta ]; then
+    cff=$outdir/$(basename $cff).reann.WITH_SEQ
+    echo cff $cff
+else
+    cff=$outdir/$(basename $cff).reann.NO_SEQ
+    echo cff $cff
+fi
+
+echo Add adjacent exons to cff
+extract_closest_exons.py $cff $gene_bed $genome_fasta  > $outdir/$(basename $cff).exons
+
+# assign cff as ".exons" if --annotate_exons flag was specified
+
+cff=$outdir/$(basename $cff).exons
+
+
+#Merge
+cluster=$outdir/$(basename $cff).cluster
+echo Merge cff by genes and breakpoints
+RUN_cluster_genes_breakpoints.sh $cff $outdir > $cluster
+
+#output ANC_RT_SG file
+if [ $output_ANC_RT_SG -eq 1 ]; then
+    echo output cis-sage.cluster file
+    output_ANC_RT_SG.py $cluster > $outdir/cis-sage.cluster
+fi
+
+#ReadThrough Callerfilter
+if [ $RT_call_filter -eq 1 ]; then
+    echo ReadThrough, callerfilter $num_tools
+    cat $cluster | grep ReadThrough > $outdir/$(basename $cluster).ReadThrough
+    callerfilter_num.py --cluster $cluster  --num_tools $num_tools > $outdir/$(basename $cluster).callerfilter.$num_tools
+    callerfilter_excluded=$(comm -13 <(cut -f 22 $outdir/$(basename $cluster).callerfilter.$num_tools | sort | uniq) <(cut -f 22 $cluster | sort | uniq))
+    grep -v ReadThrough $outdir/$(basename $cluster).callerfilter.$num_tools > $outdir/$(basename $cluster).RT_filter.callerfilter.$num_tools
+    cluster_RT_call=$outdir/$(basename $cluster).RT_filter.callerfilter.$num_tools
+fi
+# Blocklist Filter
+if [ $recurrent_bedpe ]; then
+    echo blocklist filter
+    blocklist_filter_recurrent_breakpoints.sh $cff $cluster_RT_call $outdir $recurrent_bedpe > $outdir/$(basename $cluster).RT_filter.callerfilter.$num_tools.blck_filter
+
+    blocklist_cluster=$outdir/$(basename $cluster_RT_call).BLOCKLIST
+
+    cluster=$outdir/$(basename $cluster).RT_filter.callerfilter.$num_tools.blck_filter
+fi
+# Adjacent Noncoding filter
+if [ $ANC_filter -eq 1 ]; then
+    echo ANC adjacent noncoding filter
+    filter_adjacent_noncoding.py $cluster > $outdir/$(basename $cluster).ANC_filter
+
+    cluster=$outdir/$(basename $cluster).ANC_filter
+fi
+#Rank and generate final.cluster
+echo Rank and generate final.cluster
+rank_cluster_file.py $cluster > $outdir/final.n$num_tools.cluster
+cluster=$outdir/final.n$num_tools.cluster
+### Generate filtered FID file
+#out=`awk -F '\t' '{print $15}' $cluster  | tail -n +2`
+#out2=`awk -F '\t' '{print $22}' $outdir/cis-sage.cluster | tail -n +2`
+#out3=`echo $out $out2`
+#echo ${out3//,/ } > out4
+#out5=`tr ' ' '\n' < out4 | sort | uniq`
+
+#for this in $(echo $out5); do grep $this $cff; done >> $outdir/$(basename $cff).filtered.cff
+
+rm -f filters.txt
+cut -f 22 *.BLOCKLIST | tr "," "\n" | sort | uniq | sed "s/$/\tblocklist/g" > filters.txt
+cut -f 22 *.ANC_filter | tr "," "\n" | sort | uniq | sed "s/$/\tadjacent_noncoding/g" >> filters.txt
+cut -f 22 *.ReadThrough | tr "," "\n" | sort | uniq | sed "s/$/\tread_through/g" >> filters.txt
+echo -en "$callerfilter_excluded" | tr "," "\n" | sort | uniq | sed "s/$/\tcaller_filter/g" >> filters.txt
+

bin/add_annotations_cff.R

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+#!/usr/local/bin/Rscript
+# __author__      = "Anne Marie Noronha"
+# __email__       = "[email protected]"
+# __version__     = "0.0.1"
+
+
+suppressPackageStartupMessages({
+    library(dplyr)
+    library(data.table)
+})
+
+usage <- function() {
+    message("Usage:")
+    message("add_annotations_cff.R --cff-file <file.cff> --agfusion-file <agfusion.tsv> --oncokb-file <oncokb.tsv> --out-prefix <prefix>")
+}
+
+args = commandArgs(TRUE)
+
+if (is.null(args) | length(args)<1) {
+    usage()
+    quit()
+}
+
+#' Parse out options from a string without recourse to optparse
+#'
+#' @param x Long-form argument list like --opt1 val1 --opt2 val2
+#'
+#' @return named list of options and values similar to optparse
+
+parse_args <- function(x){
+    args_list <- unlist(strsplit(x, ' ?--')[[1]])[-1]
+    args_vals <- lapply(args_list, function(x) scan(text=x, what='character', quiet = TRUE))
+    # Ensure the option vectors are length 2 (key/ value) to catch empty ones
+    args_vals <- lapply(args_vals, function(z){ length(z) <- 2; z})
+
+    parsed_args <- structure(lapply(args_vals, function(x) x[2]), names = lapply(args_vals, function(x) gsub('-','_',x[1])))
+    parsed_args[! is.na(parsed_args)]
+}
+
+args_opt <- parse_args(paste(args,collapse=" "))
+
+possible_args = c("cff", "oncokb", "agfusion", "out_prefix")
+if (length(setdiff(names(args_opt),possible_args)) > 0){
+    message("Invalid options")
+    usage()
+    quit()
+}
+
+if (length(setdiff(possible_args,names(args_opt))) > 0) {
+    message("Missing required arguments")
+    usage()
+    quit()
+}
+
+oncokb_file = args_opt$oncokb
+agfusion_file = args_opt$agfusion
+cff_file = args_opt$cff
+out_prefix = args_opt$out_prefix
+
+cff = fread(cff_file)
+final_cff_cols <- c(names(cff))
+agfusion_tab = fread(agfusion_file) %>% select(c(`5'_transcript`,`3'_transcript`,`5'_breakpoint`,`3'_breakpoint`,Fusion_effect))
+final_cff_cols <- c(final_cff_cols,"Fusion_effect")
+if (!is.null(oncokb_file)){
+    oncokb_tab = fread(oncokb_file) %>% select(-Fusion)
+    final_cff_cols = c(final_cff_cols,names(oncokb_tab %>% select(-Tumor_Sample_Barcode)))
+    cff <- merge(
+        cff,
+        oncokb_tab,
+        by.x ="FID",
+        by.y = "Tumor_Sample_Barcode",
+        all.x = T,
+        all.y=F
+    )
+}
+
+cff <- merge(
+    cff,
+    agfusion_tab,
+    by.x = c("gene5_transcript_id","gene3_transcript_id","gene5_breakpoint","gene3_breakpoint"),
+    by.y = c("5'_transcript","3'_transcript","5'_breakpoint","3'_breakpoint"),
+    all.x = T,
+    all.y = T
+)
+
+cff <- as.data.frame(cff)[,c(final_cff_cols)]
+#cff <- cff %>% mutate(!!final_cff_cols[34] := Fusion_effect) %>% select(-c(Fusion_effect))
+
+write.table(
+    cff,
+    paste0(out_prefix, ".unfiltered.cff"),
+    row.names = F,
+    quote = F,
+    sep = "\t",
+    col.names = ! "V1" %in% final_cff_cols
+)
+
+filtered_cff <- cff %>% filter(! (is.na(cluster) | is.null(cluster) | cluster == ""))
+write.table(
+    filtered_cff,
+    paste0(out_prefix, ".final.cff"),
+    row.names = F,
+    append = F,
+    quote = F,
+    sep = "\t",
+    col.names = ! "V1" %in% final_cff_cols
+)
+
+