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#' Get indel contexts
#'
#' @details
#' Determines the COSMIC context from a GRanges or GRangesList object containing Indel mutations.
#' It applies the get_indel_context_gr function to each gr in the input.
#' It searches for repeat units both to the left and right of the indel.
#'
#' @param vcf_list GRanges or GRangesList object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input grl. In each gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is either the number of repeats or the microhomology length.
#'
#' @examples
#'
#' ## Get a GRangesList or GRanges object with only indels.
#' ## See 'read_vcfs_as_granges' or 'get_mut_type' for more info on how to do this.
#' indel_grl <- readRDS(system.file("states/blood_grl_indel.rds",
#' package = "MutationalPatterns"
#' ))
#'
#' ## Load the corresponding reference genome.
#' ref_genome <- "BSgenome.Hsapiens.UCSC.hg19"
#' library(ref_genome, character.only = TRUE)
#'
#' ## Get the indel contexts
#' get_indel_context(indel_grl, ref_genome)
#' @family Indels
#' @importFrom magrittr %>%
#' @seealso
#' \code{\link{read_vcfs_as_granges}}, \code{\link{get_mut_type}}
#'
#' @export
#'
get_indel_context <- function(vcf_list, ref_genome) {
# Check that the seqnames of the gr and ref_genome match
.check_chroms(vcf_list, ref_genome)
# Convert list to grl if necessary
if (inherits(vcf_list, "list")) {
vcf_list <- GenomicRanges::GRangesList(vcf_list)
}
if (inherits(vcf_list, "CompressedGRangesList")){
# Unlist the GrangesList into a single GRanges object.
gr <- BiocGenerics::unlist(vcf_list, use.names = FALSE)
# Link samples to mutations
nr_muts <- S4Vectors::elementNROWS(vcf_list)
gr$INTERNAL_SAMPLENAME <- rep(names(nr_muts), times = nr_muts)
# Get mutation context
gr <- .get_indel_context_gr(gr, ref_genome)
# Split the GRanges back into a list.
sample_indx <- gr$INTERNAL_SAMPLENAME
gr$INTERNAL_SAMPLENAME <- NULL
grl <- S4Vectors::split(gr, sample_indx)
return(grl)
} else if (inherits(vcf_list, "GRanges")) {
gr <- .get_indel_context_gr(vcf_list, ref_genome)
return(gr)
} else {
.not_gr_or_grl(vcf_list)
}
}
#' Get indel contexts from a single gr
#'
#' @details
#' Determines the COSMIC context from a GRanges object containing Indel mutations.
#' It throws an error if there are any variants with multiple alternative alleles or SNVs.
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input gr. In the gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is either the number of repeats or the microhomology length.
#'
#'
#' @noRd
#'
#' @importFrom magrittr %>%
#'
.get_indel_context_gr <- function(gr, ref_genome) {
# Check that no snvs are present.
.check_no_substitutions(gr)
# Calculate indel size to determine main category
ref_sizes <- gr %>%
.get_ref() %>%
width()
alt_sizes <- gr %>%
.get_alt() %>%
unlist() %>%
width()
mut_size <- alt_sizes - ref_sizes
# For the main indel categories, determine their sub categories.
# (Also split the big deletion categorie into repeat and micro homology.)
gr_1b_dels <- .get_1bp_dels(gr, mut_size, ref_genome)
gr_1b_ins <- .get_1bp_ins(gr, mut_size, ref_genome)
gr_big_dels <- .get_big_dels(gr, mut_size, ref_genome)
gr_big_ins <- .get_big_ins(gr, mut_size, ref_genome)
gr <- c(gr_1b_dels, gr_1b_ins, gr_big_dels, gr_big_ins) %>%
BiocGenerics::sort()
return(gr)
}
#' Get contexts from 1bp deletions
#'
#' @details
#' Determines the COSMIC context for the 1bp deletions in a GRanges object containing Indel mutations.
#' This function is called by get_indel_context_gr.
#'
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param mut_size A double vector containing the size of each Indel.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input gr.
#' All variants that are not a 1bp deletion are removed.
#' In each gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is the number of repeats.
#'
#'
#' @importFrom magrittr %>%
#' @noRd
#'
#'
.get_1bp_dels <- function(gr, mut_size, ref_genome) {
# Select mutations
gr <- gr[mut_size == -1]
if (length(gr) == 0) {
return(gr)
}
# Get the deleted bases
changed_bases <- gr %>%
.get_ref() %>%
as.character() %>%
substring(2)
# Remove any potential names from the ALT column so str_replace_all will work
names(changed_bases) <- NULL
# Get homopolymer length
homopolymer_length <- .indel_get_n_repeats(gr, 19, ref_genome, changed_bases, "deletion")
changed_bases[changed_bases == "A"] <- "T"
changed_bases[changed_bases == "G"] <- "C"
# Return the results
gr$muttype <- stringr::str_c(changed_bases, "_deletion")
gr$muttype_sub <- homopolymer_length
return(gr)
}
#' Get contexts from 1bp insertions
#'
#' @details
#' Determines the COSMIC context for the 1bp insertions in a GRanges object
#' containing Indel mutations. This function is called by get_indel_context_gr.
#'
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param mut_size A double vector containing the size of each Indel.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input gr.
#' All variants that are not a 1bp insertion are removed.
#' In each gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is the number of repeats.
#'
#'
#' @importFrom magrittr %>%
#' @noRd
#'
.get_1bp_ins <- function(gr, mut_size, ref_genome) {
# Select mutations
gr <- gr[mut_size == 1]
if (length(gr) == 0) {
return(gr)
}
# Get inserted bases.
changed_bases <- gr %>%
.get_alt() %>%
unlist() %>%
as.character() %>%
substring(2)
# Remove any potential names from the ALT column so str_replace_all will work
names(changed_bases) <- NULL
# Get homopolymer length
homopolymer_length <- .indel_get_n_repeats(gr, 20, ref_genome, changed_bases, "insertion")
changed_bases[changed_bases == "A"] <- "T"
changed_bases[changed_bases == "G"] <- "C"
# Return the results
gr$muttype <- stringr::str_c(changed_bases, "_insertion")
gr$muttype_sub <- homopolymer_length
return(gr)
}
#' Get contexts from bigger inserions
#'
#' @details
#' Determines the COSMIC context for insertions larger than 1bp in a GRanges
#' object containing Indel mutations. This function is called by
#' get_indel_context_gr.
#'
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param mut_size A double vector containing the size of each Indel.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input gr.
#' All variants that are not insertions larger than 1bp are removed.
#' In each gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is the number of repeats.
#'
#'
#' @importFrom magrittr %>%
#' @noRd
#'
.get_big_ins <- function(gr, mut_size, ref_genome) {
# Select mutations
gr <- gr[mut_size > 1]
if (length(gr) == 0) {
return(gr)
}
mut_size <- mut_size[mut_size > 1]
# Get inserted bases
changed_bases <- gr %>%
.get_alt() %>%
unlist() %>%
as.character() %>%
substring(2)
biggest_ins <- changed_bases %>%
nchar() %>%
max()
flank_dist <- biggest_ins * 20
# Remove any potential names from the ALT column so str_replace_all will work
names(changed_bases) <- NULL
# Get number of repeats
n_repeats <- .indel_get_n_repeats(gr, flank_dist, ref_genome, changed_bases, "insertion")
# Return results
gr$muttype <- stringr::str_c(mut_size, "bp_insertion")
gr$muttype_sub <- n_repeats
return(gr)
}
#' Get contexts from larger deletions
#'
#' @details
#' Determines the COSMIC context for deletions larger than 1bp in a GRanges object
#' containing Indel mutations.
#' This function is called by get_indel_context_gr.
#' The function determines if there is microhomology for deletions that are not in repeat regions.
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param mut_size A double vector containing the size of each Indel.
#' @param ref_genome BSgenome reference genome object
#'
#' @return A modified version of the input gr.
#' All variants that are not a deletion larger than 1bp are removed.
#' In each gr two columns have been added.
#' "muttype" showing the main indel type and "muttype_sub" which shows the subtype.
#' The subtype is the number of repeats or the microhomology length.
#'
#'
#' @importFrom magrittr %>%
#' @noRd
#'
.get_big_dels <- function(gr, mut_size, ref_genome) {
# Select mutations
gr <- gr[mut_size < -1]
if (length(gr) == 0) {
return(gr)
}
mut_size <- mut_size[mut_size < -1]
# Get deleted bases
changed_bases <- gr %>%
.get_ref() %>%
as.character() %>%
substring(2)
biggest_dels <- changed_bases %>%
nchar() %>%
max()
flank_dist <- biggest_dels * 20
# Remove any potential names from the ALT column so str_replace_all will work
names(changed_bases) <- NULL
# Get number of repeats
n_repeats <- .indel_get_n_repeats(gr, flank_dist, ref_genome, changed_bases, "deletion")
gr$muttype <- stringr::str_c(abs(mut_size), "bp_deletion")
gr$muttype_sub <- n_repeats
# Determine if there is microhomology for deletions that are not in repeat regions.
# There is always at least 1 'repeat', because of the deleted bases themselves.
pos_mh <- gr$muttype_sub == 1
gr_repeat <- gr[!pos_mh]
gr_mh <- gr[pos_mh]
if (length(gr_mh) == 0) {
return(gr_repeat)
}
mut_size_mh <- mut_size[pos_mh]
del_bases_mh <- changed_bases[pos_mh]
del_bases_s <- strsplit(del_bases_mh, "")
seq <- .get_extended_sequence(gr_mh, biggest_dels, ref_genome, "right")
seq_s <- strsplit(as.character(seq), "")
# Also check for microhomology to the left of the deletion.
# For this take the reverse sequence to the left
# of the deletion and the reverse deleted bases.
rev_del_bases <- Biostrings::reverse(del_bases_mh)
rev_l_del_bases_s <- strsplit(rev_del_bases, "")
seq_left <- .get_extended_sequence(gr_mh, biggest_dels, ref_genome, "left")
rev_l_seq_s <- strsplit(as.character(seq_left), "")
# For each mutation determine how many bases show hm
nr_pos_mh <- length(del_bases_s)
nr_mh <- vector("list", nr_pos_mh)
for (i in seq_len(nr_pos_mh)) {
del_bases_sample <- del_bases_s[[i]]
seq_s_sample <- seq_s[[i]][seq_len(length(del_bases_sample))]
same <- del_bases_sample == seq_s_sample
# Determine how many bases are the same before the first difference.
# na.rm is for when a sequence has been trimmed.
r_nr_mh_sample <- cumprod(same) %>%
sum(na.rm = TRUE)
l_del_bases_sample <- rev_l_del_bases_s[[i]]
l_seq_s_sample <- rev_l_seq_s[[i]][seq_len(length(l_del_bases_sample))]
l_same <- l_del_bases_sample == l_seq_s_sample
l_nr_mh_sample <- cumprod(l_same) %>%
sum(na.rm = TRUE)
nr_mh_sample <- max(r_nr_mh_sample, l_nr_mh_sample)
nr_mh[[i]] <- nr_mh_sample
}
nr_mh <- unlist(nr_mh)
# Update gr when mh is indeed present
mh_f <- nr_mh > 0
gr_mh$muttype[mh_f] <- stringr::str_c(
abs(mut_size_mh[mh_f]),
"bp_deletion_with_microhomology"
)
gr_mh$muttype_sub[mh_f] <- nr_mh[mh_f]
# Combine muts with and without mh
gr <- c(gr_mh, gr_repeat) %>%
BiocGenerics::sort()
return(gr)
}
#' Count how often an indel is repeated on both the left
#' and the right side of the indel. Takes the side with
#' the maximum number of repeats.
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param flank_dist A numeric vector of length one containing
#' the number of flanking base pairs.
#' @param ref_genome BSgenome reference genome object
#' @param changed_bases String of the mutated bases.
#' @param type Indel type. Either deletion or insertion.
#'
#' @return double vector containing the number of repeats per indel.
#' @noRd
#'
.indel_get_n_repeats = function(gr, flank_dist, ref_genome, changed_bases, type){
# Determine the number of repeats for both the left and the right flank.
n_repeats_right <- .indel_get_n_repeats_right(gr, flank_dist, ref_genome, changed_bases, type)
n_repeats_left <- .indel_get_n_repeats_left(gr, flank_dist, ref_genome, changed_bases, type)
n_repeats <- pmax(n_repeats_right, n_repeats_left)
return(n_repeats)
}
#' Count how often an indel is repeated to the right
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param flank_dist A numeric vector of length one containing
#' the number of flanking base pairs.
#' @param ref_genome BSgenome reference genome object
#' @param changed_bases String of the mutated bases.
#' @param type Indel type. Either deletion or insertion.
#'
#' @return double vector containing the number of repeats per indel.
#' @noRd
#'
.indel_get_n_repeats_right = function(gr, flank_dist, ref_genome, changed_bases, type){
# Find extended sequence
seq <- .get_extended_sequence(gr, flank_dist, ref_genome, direction = "right")
n_repeats <- .indel_count_n_repeats(seq, changed_bases, type)
return(n_repeats)
}
#' Count how often an indel is repeated to the left.
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param flank_dist A numeric vector of length one containing
#' the number of flanking base pairs.
#' @param ref_genome BSgenome reference genome object
#' @param changed_bases String of the mutated bases.
#' @param type Indel type. Either deletion or insertion.
#'
#' @return double vector containing the number of repeats per indel.
#' @noRd
#'
.indel_get_n_repeats_left = function(gr, flank_dist, ref_genome, changed_bases, type){
# Find extended sequence
seq <- .get_extended_sequence(gr, flank_dist, ref_genome, direction = "left")
# Reverse the bases, to look in the left direction.
changed_bases <- Biostrings::reverse(changed_bases)
n_repeats <- .indel_count_n_repeats(seq, changed_bases, type)
return(n_repeats)
}
#' Retreive the flanking sequence from a GRanges object
#'
#' @details
#' This function retreives the flanking sequence from a GRanges object.
#' The size of the flanking sequence is supplied as an argument to the function.
#' This function works by first extending the ranges and then retreiving the sequence.
#'
#'
#'
#' @param gr GRanges object containing Indel mutations.
#' The mutations should be called similarly to HaplotypeCaller.
#' @param flank_dist A numeric vector of length one containing the number of flanking base pairs.
#' @param ref_genome BSgenome reference genome object
#' @param direction Whether to flank to the right or left of the indel
#' @return A DNAStringSet containing the flanking bases.
#'
#' @noRd
#'
.get_extended_sequence <- function(gr, flank_dist, ref_genome, direction = c("right", "left")) {
# Match argument
direction <- match.arg(direction)
if (direction == "right"){
# Flank the granges object on the right, to get a sequence, that can be
# searched for repeats. This can result in a warning message, when the
# flanked range extends beyond the chrom length. This message is suppressed.
withCallingHandlers(
{
gr_extended <- GenomicRanges::flank(gr, flank_dist, start = FALSE)
},
warning = function(w) {
if (grepl("out-of-bound range located on sequence", conditionMessage(w))) {
invokeRestart("muffleWarning")
}
}
)
} else{
# Flank the granges object on the left, to get a sequence, that can be
# searched for repeats. This can result in a warning message, when the
# flanked range extends beyond the chrom length. This message is suppressed.
withCallingHandlers(
{
gr_extended <- GenomicRanges::flank(gr, flank_dist-1, start = TRUE)
BiocGenerics::end(gr_extended) <- BiocGenerics::end(gr_extended) + 1
},
warning = function(w) {
if (grepl("out-of-bound range located on sequence", conditionMessage(w))) {
invokeRestart("muffleWarning")
}
}
)
}
# Trim the ranges that are extended beyond the actual length of the chromosome.
gr_extended <- GenomicRanges::trim(gr_extended)
seq <- Biostrings::getSeq(BSgenome::getBSgenome(ref_genome), gr_extended)
#Reverse the sequence if its retreived at the left.
if (direction == "left"){
seq <- Biostrings::reverse(seq)
}
return(seq)
}
#' Counts how often the mutated bases of an indel are repeated
#' in the surrounding context.
#'
#' @param seq A DNAStringSet containing the flanking bases.
#' @param changed_bases String of the mutated bases.
#' @param type Indel type. Either deletion or insertion.
#'
#' @return double vector containing the number of repeats per indel.
#' @noRd
#'
.indel_count_n_repeats = function(seq, changed_bases, type = c("deletion", "insertion")){
# Match argument
type <- match.arg(type)
# Determine nr. repeats.
# For each mut replace the deleted basetype in the flanking sequence with Zs.
seq_z <- stringr::str_replace_all(
as.character(seq),
changed_bases,
rep("Z", length(seq))
)
# Remove all bases after the Zs and count how many bases are left.
# Add +1 for the deleted bases itself
n_repeats <- gsub("[^Z].*", "", as.character(seq_z)) %>%
nchar()
if (type == "deletion"){
n_repeats <- n_repeats + 1
}
return(n_repeats)
}
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