R/finalRegionFunctions.R

Defines functions .loadPeaks .convertSallToGrl findOverlapsOverSamples initMergedPeaksNames giveUniqueNamesToPeaksOverSamples finalRegions

Documented in finalRegions findOverlapsOverSamples giveUniqueNamesToPeaksOverSamples initMergedPeaksNames

#' finalRegions
#' @description Align peaks to form common regions then filter regions for
#' presence in multiple replicates taking in input a GRangesList where each
#' element is a sample of called peaks.
#'
#' @param peakSamplesGRangesList named GRangesList where each element is a
#' sample of called peaks. A score mcols values is needed for each GRanges.
#' The scorecolname param can be used as reference name for the score.
#' (tipically returned by findPeaks function).
#' @param zThreshold a minimum threshold for the z score.
#' All peaks lesser than this value will be ignored.
#' @param minCarriers a threshold of minimum samples (carriers) for overlapped
#'                    regions.
#' @param saveFlag a flag for saving results in a tsv file.
#' @param outputFolder the directory name to store the bed file.
#' @param verbose verbose output.
#' @param scorecolname character describing the name of the column within the
#' peaks score.
#' @param BPPARAM object of class \code{bpparamClass} that specifies the
#'   back-end to be used for computations. See
#'   \code{\link[BiocParallel]{bpparam}} for details.
#'
#' @return a GRanges of selected overlapping peaks with z-score,
#' n-peaks, k-carriers as mcols object.
#' @export
#' @importFrom GenomicRanges GRangesList
#' @importFrom S4Vectors mcols
#' @importFrom BiocParallel bpparam bplapply
#' @examples
#' peak.path <- system.file("extdata/peaks/RData/peaksGRL_all_files.rds",
#'                             package="DEScan2")
#' grl <- readRDS(peak.path)
#' grl
#'
#' regionsGR <- finalRegions(peakSamplesGRangesList=grl, zThreshold=1,
#'                         minCarriers=3, saveFlag=FALSE, verbose=TRUE)
finalRegions <- function(peakSamplesGRangesList, zThreshold=20, minCarriers=2,
                                    saveFlag=TRUE,
                                    outputFolder="overlappedPeaks",
                                    verbose=FALSE,
                                    scorecolname="z-score",
                                    BPPARAM=BiocParallel::bpparam())
{
    stopifnot(is(peakSamplesGRangesList, "GRangesList"))

    if(verbose) message("computing final regions on ",
                        length(peakSamplesGRangesList), " samples...")
    zedPeaksSamplesGRList <- GenomicRanges::GRangesList(
                    lapply(peakSamplesGRangesList, function(sample)
                    {
                        return(sample[
                            which(S4Vectors::mcols(sample)[[scorecolname]]
                                                            >= zThreshold),])
                    }))

    zedPeaksChrsGRList <- fromSamplesToChrsGRangesList(zedPeaksSamplesGRList)

    #### to parallelize over chrs
    overlappedPeaksGRList <- GenomicRanges::GRangesList(
        BiocParallel::bplapply(zedPeaksChrsGRList, function(chrSampleGRList)
        {
            return(findOverlapsOverSamples(chrSampleGRList,
                    zThresh=zThreshold, verbose=verbose,
                    scorecolname=scorecolname))
        }, BPPARAM=BPPARAM)
    )
    overlappedPeaksGR <- unlist(overlappedPeaksGRList)
    idxK <- which(overlappedPeaksGR$`k-carriers` >= minCarriers)
    overlMinKPeaksGR <- overlappedPeaksGR[idxK,]

    if(saveFlag)
    {
        datename <- paste0(strsplit(gsub(pattern=":", replacement=" ",
                                            date()), " ")[[1]], collapse="_")
        filename <- paste0("regions_", datename, "_zt", zThreshold,
                            "_minK", minCarriers)
        saveGRangesAsTsv(GRanges=overlMinKPeaksGR, filepath=outputFolder,
                        filename=filename, verbose=verbose)
    }
    return(overlMinKPeaksGR)
}

#' giveUniqueNamesToPeaksOverSamples
#' @description given a GRangesList of samples assigns unique names to the peaks
#' of each sample.
#' @param samplePeaksGRangelist a GRangeList of peaks, one GRanges for each
#' sample.
#'
#' @return a GRangesList of samples within renamed peaks for each element.
#' @keywords internal
giveUniqueNamesToPeaksOverSamples <- function(samplePeaksGRangelist)
{
    stopifnot(is(samplePeaksGRangelist, "GRangesList"))
    ## this is just for naming the peaks
    ## total number of samples
    ns <- length(samplePeaksGRangelist)
    ## total number of decimals for the samples
    ncs <- nchar(as.character(ns))
    ## total number of decimals for the peaks taking the max number of peaks
    ncp <- nchar(as.character(max(unlist(
                                        lapply(samplePeaksGRangelist, length)
                                        ))))
    sFormat <- paste0("s%0", ncs,"d")
    format <- paste0("s%0", ncs,"d_p%0", ncp, "d")
    listNames <- character()
    ## for each sample assign unique names to the peaks
    samplePeaksGRangelista <- lapply(
        seq_along(samplePeaksGRangelist),
        function(x, i)
        {
            peakNames <- sprintf(format, i, seq_len(length(x[[i]])))
            # print(peakNames)
            y <- x[[i]]
            names(y) <- peakNames
            return(y)
            # x[[i]] <- y
            # names(x[[i]]) <- peakNames
            # return(x[[i]])
        },
        x=samplePeaksGRangelist
    )

    names(samplePeaksGRangelista) <- sprintf(sFormat,
                                            seq_along(samplePeaksGRangelist))
    return(samplePeaksGRangelista)
}

#' initMergedPeaksNames
#' @description given a GRanges of merged peaks assigns them new names.
#' @param mergedGRanges A GRanges object.
#' (Tipically Generated in findOverlapsOverSamples function )
#'
#' @return a granges of renamed peaks.
#' @keywords internal
initMergedPeaksNames <- function(mergedGRanges)
{
    stopifnot(is(mergedGRanges, "GRanges"))
    ncp <- nchar(length(mergedGRanges@ranges))
    nk <- nchar(max(mergedGRanges$`k-carriers`))
    np <- nchar(max(mergedGRanges$`n-peaks`))
    format <-  paste0("p%0", ncp, "d_np%0", np, "d_k%0", nk,"d")

    peakNames <- sprintf(format, seq_len(length(mergedGRanges@ranges)),
                        mergedGRanges$`n-peaks`, mergedGRanges$`k-carriers`)
    names(mergedGRanges) <- peakNames
    return(mergedGRanges)
}

#' findOverlapsOverSamples
#' @description given in input a GRangeList where each element is a sample
#' computes the coverage extending a both direction window of prefixed length.
#'
#' @param samplePeaksGRangelist given a granges list of samples finds
#' the overlapping regions between them.
#' @param extendRegions the number of bases to extend each region at its start
#' and end.
#' @param minOverlap the minimum overlap each peak needs to have.
#' (see ChipPeakAnno::findOverlapsOfPeaks)
#' @param maxGap the maximum gap admissible between the peaks.
#' (see ChipPeakAnno::findOverlapsOfPeaks)
#' @param verbose verbose flag
#' @param scorecolname character describing the name of the column within the
#' peaks score.
#' @param zThresh a threshold value on z-score/scorecolname
#'
#' @return a GRanges of peaks overlapped and unique between samples.
#' @export
#'
#' @importFrom S4Vectors mcols runValue
#' @importFrom BiocGenerics start end
#' @importFrom GenomeInfoDb seqlengths seqnames
#' @importFrom ChIPpeakAnno findOverlapsOfPeaks
#' @importFrom data.table rbindlist
#' @importFrom GenomicRanges GRangesList
#'
#' @examples
#' (peaks.file <- system.file("extdata/peaks/RData/peaksGRL_all_files.rds",
#'                             package="DEScan2"))
#' peaksGRLFiles <- readRDS(peaks.file)
#' (overlPeaks <- findOverlapsOverSamples(peaksGRLFiles))
findOverlapsOverSamples <- function(samplePeaksGRangelist,
                                    extendRegions=200,
                                    minOverlap=0L,
                                    maxGap=-1L,
                                    zThresh=10,
                                    verbose=FALSE,
                                    scorecolname="z-score")
{
    stopifnot(is(samplePeaksGRangelist, "GRangesList"))

    namedSamplePeaksGRL <- giveUniqueNamesToPeaksOverSamples(
                                                        samplePeaksGRangelist)

    namedSamplePeaksGRL <- lapply(namedSamplePeaksGRL, function(x)
    {
        x <- x[as.numeric(S4Vectors::mcols(x)[[scorecolname]]) >= zThresh]
        if(length(x) ==0 ) stop("no peaks found in one sample\n",
                            "please try again providing a lower zThreshold")
        S4Vectors::mcols(x)[["n-peaks"]] <-  1
        S4Vectors::mcols(x)[["k-carriers"]] <-  1
        BiocGenerics::start(x) <- BiocGenerics::start(x) - extendRegions
        idxNeg <- which( BiocGenerics::start(x) < 0 )
        if(length(idxNeg) > 0)
        {
            warning("extendRegions of ", extendRegions,
                    " is too high for region(s) start ", idxNeg,
                    " forcing these starts to 0")
            BiocGenerics::start(x)[idxNeg] <- 0
        }
        BiocGenerics::end(x) <- BiocGenerics::end(x) + extendRegions
        ## it must return just one chromosome
        idxHigh <- which( BiocGenerics::end(x) > GenomeInfoDb::seqlengths(x) )
        if(length(idxHigh) > 0) {
            warning("extendRegions of ", extendRegions,
                    " is too high for region(s) end ", idxHigh,
                    " forcing these ends to ", GenomeInfoDb::seqlengths(x))
            BiocGenerics::end(x)[idxHigh] <- GenomeInfoDb::seqlengths(x)
        }

        return(x)
    })


    if(verbose) message("Computing overlapping regions over samples...")
    # startTime <- Sys.time()
    if(length(namedSamplePeaksGRL) > 1 )
    {
        for(i in 2:length(namedSamplePeaksGRL))
        {
            if( i == 2 ) {
                gri <- namedSamplePeaksGRL[[1]]
                foundedPeaks <- NULL
            } else {
                gri <- foundedPeaks
            }

            grj <- namedSamplePeaksGRL[[i]]

            grij <- ChIPpeakAnno::findOverlapsOfPeaks(gri,
                                                        grj,
                                                        minoverlap=minOverlap,
                                                        maxgap=maxGap,
                                                        connectedPeaks="merge")

            mmpeaks <- grij$peaksInMergedPeaks
            if(length(mmpeaks) == 0)
            {
                message("No merged peaks found at sample ", i,
                    " and chromosome ",
                    as.character(S4Vectors::runValue(
                        GenomeInfoDb::seqnames(grj))),
                    "\nNB: skipping this sample!")
                foundedPeaks <- gri
                next
            }

            ## cleaning peaks names
            mrgPks <- grij$mergedPeaks
            mrgPksNms <- as.list(mrgPks$peakNames)
            # stTime <- Sys.time()
            newcols <- lapply(mrgPksNms, function(l)
            {
                idx <- which(names(mmpeaks) %in% l)
                scores <- as.numeric(S4Vectors::mcols(mmpeaks)[idx,
                                                            scorecolname])
                nPeaks <- as.numeric(mmpeaks$`n-peaks`[idx])
                kCarr <- as.numeric(mmpeaks$`k-carriers`[idx])
                np = sum(nPeaks) ## total number of peaks found
                ## it's necessary to rescale the score on the basis of the peaks
                ## found from previous computations
                mmzp <- sum(scores*nPeaks)/np
                ## the carriers are just the number of samples
                k <- max(kCarr)+1
                as.data.frame(cbind(mmzp, np, k))
            })
            # endTime <- Sys.time()
            # print((endTime-stTime))
            newcols1 <- data.table::rbindlist(newcols)
            S4Vectors::mcols(mrgPks) <-  S4Vectors::DataFrame(newcols1)
            if( dim(S4Vectors::mcols(mrgPks))[1] == 0 )
            {
                stop("No overlapping regions found!")
            }
            colnames(S4Vectors::mcols(mrgPks)) <- c(scorecolname,
                                                        "n-peaks",
                                                        "k-carriers")

            ## peaks uniques
            unqPks <- grij$uniquePeaks
            ## putting together all the peaks
            foundedPeaks <- unlist(GenomicRanges::GRangesList(unqPks, mrgPks))

            foundedPeaks <- initMergedPeaksNames(foundedPeaks)
        }
    } else if(length(namedSamplePeaksGRL) == 1) {
        foundedPeaks <- initMergedPeaksNames(namedSamplePeaksGRL[[1]])
    } else if(length(namedSamplePeaksGRL) == 0){
        stop("No regions found with this threshold!")
    }
    # endingTime <- Sys.time()
    # print((endingTime - startTime))
    # save(foundedPeaks, file="testData/new_files/foundedPeaks.RData")
    return(foundedPeaks)
}


.convertSallToGrl <- function(sall)
{
    lgr <- list()
    for(sample in sall)
    {
        gr <- GenomicRanges::GRanges(seqnames=sample[,1],
                        ranges=IRanges::IRanges(start=as.numeric(sample[,2]),
                                                end=as.numeric(sample[,3])
                                                )
        )
        S4Vectors::mcols(gr)[["z-score"]] <- as.numeric(sample[,4])
        lgr <- c(lgr, gr)
    }

    grl <- GenomicRanges::GRangesList(lgr)
    return(grl)
}

.loadPeaks <- function(peakdirname, verbose = FALSE)
{
    files <- list.files(peakdirname, pattern="Peaks", full.names=TRUE)
    if (verbose)
    {
        message("Found", length(files), "peak files.\n")
    }
    peaks_all <- list()

    for (i in seq_len(length(files)))
    {
        p = load(files[i])
        if(length(p) > 1 )
        {
            peaksi <- eval(expr=parse(text=paste0("peaksi <- ", p[1])))
        } else {
            peaksi <- eval(parse(text=paste0("peaksi <- ", p)))
        }
        if (ncol(peaksi) == 3) {
            chr <- strsplit(peakdirname, split = "/")[[1]][2]
            peaksi <- cbind(rep(chr, nrow(peaksi)), peaksi)
        }
        peaks_all[[files[[i]]]] <- peaksi
        if (verbose) {
            message("File: ", files[i],
                    " number of regions:", nrow(peaksi), "\n")
        }
    }
    return(peaks_all)
}

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DEScan2 documentation built on Nov. 8, 2020, 5:01 p.m.