From 1b53b2a5c71a32812dadca6e5d8049ba306b15e0 Mon Sep 17 00:00:00 2001 From: Daniel Cameron Date: Wed, 22 Apr 2020 13:08:08 +1000 Subject: [PATCH] Independent reanalysis based on available raw vcfs --- ...alternate_figure2_size_binnedhistogram.pdf | Bin 0 -> 9084 bytes .../alternate_figure2_voilin.pdf | Bin 0 -> 25172 bytes .../plots_from_full_raw_vcfs.R | 205 ++++++++++++++++++ Scripts/reanalysis_plots/roc_by_sample.pdf | Bin 0 -> 62110 bytes Scripts/reanalysis_plots/roc_by_sizebin.pdf | Bin 0 -> 25558 bytes Scripts/reanalysis_plots/roc_overall.pdf | Bin 0 -> 39598 bytes 6 files changed, 205 insertions(+) create mode 100644 Scripts/reanalysis_plots/alternate_figure2_size_binnedhistogram.pdf create mode 100644 Scripts/reanalysis_plots/alternate_figure2_voilin.pdf create mode 100644 Scripts/reanalysis_plots/plots_from_full_raw_vcfs.R create mode 100644 Scripts/reanalysis_plots/roc_by_sample.pdf create mode 100644 Scripts/reanalysis_plots/roc_by_sizebin.pdf create mode 100644 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write(paste("Loading", sample), stderr()) + # Load GRIDSS + gr = breakpointRanges(readVcf(paste0("Data/raw_data/mouse/raw_vcf/gridss/", sample, "/", sample, ".chr19.full.1/gridss.annotated.vcf"))) + gr$sample = sample + gr$caller = "gridss" + caller_bpgr[[sample]][["gridss"]] = gr + + if (sample != "AKR_J") { # AKR_J is a 0 byte file + # Load lumpy + # lumpy filenames are inconsistent do we need to pattern match + gr = breakpointRanges(readVcf(list.files("Data/raw_data/mouse/raw_vcf/lumpy", full.names=TRUE, pattern=paste0("lumpexpress.", sample, ".*sorted.bam.modified.vcf$")))) + gr$sample = sample + gr$caller = "lumpy" + caller_bpgr[[sample]][["lumpy"]] = gr + } + + # Load delly + gr = breakpointRanges(readVcf(paste0("Data/raw_data/mouse/raw_vcf/delly.mouse/full/delly.", sample, "_chr19_sorted.bam.output.bcf.vcf"))) + gr$sample = sample + gr$caller = "delly" + caller_bpgr[[sample]][["delly"]] = gr + + # Load breakdancer + gr = breakpointRanges(readVcf(paste0("Data/raw_data/mouse/raw_vcf/breakdancer.mouse/full/", sample, "_chr19_sorted.bam.vcf"))) + gr$sample = sample + gr$caller = "breakdancer" + caller_bpgr[[sample]][["breakdancer"]] = gr + + # Load sniffles + # TODO: Add support for sniffles' TRA records (do not conform to the VCF specifications) to the StructuralVariantAnnotation package + # for this analysis, we can just delete the TRA records from the VCF since we're ignoring them anyway + #gr = breakpointRanges(readVcf(paste0("Data/raw_data/mouse/raw_vcf/sniffels/full/", sample, "_chr19_sorted.sniffles_output.vcf"))) + #gr$sample = sample + #gr$caller = "sniffles" + #caller_bpgr[[sample]][["sniffles"]] = gr +} + +annotate_calls = function(truth_gr, caller_gr, maxerrorbp=100) { + caller_gr$QUAL[is.na(caller_gr$QUAL)] = 0 + hits = findBreakpointOverlaps(caller_gr, truth_gr, maxgap=maxerrorbp, restrictMarginToSizeMultiple=0.5) + # Only the best QUAL TP is considered a TP, the remaining are considered FP + hits = as.data.frame(hits) %>% + mutate(QUAL=caller_gr$QUAL[queryHits]) %>% + group_by(subjectHits) %>% + arrange(desc(QUAL)) %>% + mutate(isBestHit=row_number() == 1) %>% + ungroup() + besthits = hits %>% filter(isBestHit) + otherhits = hits %>% filter(!isBestHit) + caller_gr$tp = FALSE + caller_gr$tp[besthits$queryHits] = TRUE + caller_gr$tpdup = FALSE + caller_gr$tpdup[otherhits$queryHits] = TRUE + caller_gr$true_length = NA_integer_ + caller_gr$true_length[hits$queryHits] = truth_gr[hits$subjectHits]$svLen + caller_gr$svtype = simpleEventType(caller_gr) + return(caller_gr) +} +# pair-wise correlation +for (sample in samples) { + for (caller1 in names(caller_bpgr[[sample]])) { + for (caller2 in names(caller_bpgr[[sample]])) { + if (!(sample == "AKR_J" & "lumpy" %in% c(caller1, caller2))) { + mcols(caller_bpgr[[sample]][[caller1]])[paste0(caller2, "_hits")] = countBreakpointOverlaps(caller_bpgr[[sample]][[caller1]], caller_bpgr[[sample]][[caller2]], maxgap=100, restrictMarginToSizeMultiple=0.5) + } + } + } +} + +# truth annotation +for (sample in samples) { + for (caller in names(caller_bpgr[[sample]])) { + caller_bpgr[[sample]][[caller]] = annotate_calls(truth_bpgr[[sample]], caller_bpgr[[sample]][[caller]]) + } +} +allgr = c(unlist(GRangesList(lapply(caller_bpgr, function(x) unlist(GRangesList(x))))), unlist(GRangesList(truth_bpgr))) +deldf = as.data.frame(allgr) %>% + filter(svtype == "DEL" & strand=="+") %>% # just take the lower breakend + mutate(svLen=abs(svLen)) %>% # most callers don't follow the specs: DELs should have -ve length + mutate(svLen > 50) %>% + mutate(sizebin=ifelse(svLen < 100, "<100", ifelse(svLen <= 1000, "100-999", "1000+"))) + +ggplot(deldf %>% + filter(tp & caller != "truth") %>% + group_by(caller, sizebin) %>% + mutate(mean_error=mean(abs(svLen) - abs(true_length)))) + + aes(x=abs(svLen) - abs(true_length)) + + facet_grid(sizebin ~ caller, scales="free") + + geom_histogram() + + geom_vline(aes(xintercept=mean_error), colour="blue", linetype="dotted") + + scale_x_continuous(limits=c(-100, 100)) + + labs(title="Alternative Figure 2: binned event size error distribution", x="error (bp)") +ggsave("alternate_figure2_size_binnedhistogram.pdf") + +ggplot(deldf %>% + filter(tp & caller != "truth") %>% + group_by(caller) %>% + mutate(mean_error=mean(abs(svLen) - abs(true_length)))) + + aes(y=abs(svLen) - abs(true_length), x=caller) + + geom_violin() + + labs(title="Alternative Figure 2: event size error distribution", y="error (bp)") +ggsave("alternate_figure2_voilin.pdf") + +rocby = function(df, ...) { + groupingCols <- quos(...) + df %>% + replace_na(list(QUAL=0)) %>% + mutate(QUAL=round(QUAL)) %>% + group_by(caller, !!!groupingCols) %>% + dplyr::select(QUAL, tp) %>% + group_by(caller, !!!groupingCols, QUAL) %>% + summarise(tp=sum(tp), ncalls=n()) %>% + group_by(caller, !!!groupingCols) %>% + arrange(desc(QUAL)) %>% + mutate(cumtp=cumsum(tp), cumncalls=cumsum(ncalls)) %>% + group_by(!!!groupingCols) %>% + mutate(total_tp=max(cumtp)) %>% # truth set is in here so it'll be the max + ungroup() %>% + mutate( + sensitivity=cumtp/total_tp, + precision=cumtp/cumncalls) +} + +overall_roc_df = bind_rows( + rocby(deldf) %>% mutate(callset="All calls"), + rocby(deldf %>% filter(FILTER %in% c("PASS", "."))) %>% mutate(callset="PASS only")) + +ggplot(overall_roc_df) + + aes(x=sensitivity, y=precision, color=caller, linetype=callset) + + geom_line() + + geom_point(data=overall_roc_df %>% group_by(caller, callset) %>% top_n(1, sensitivity), aes(shape=callset)) + + labs(title="Precision recall with QUAL and FILTER") +ggsave("roc_overall.pdf") + +ggplot(rocby(deldf, sample)) + + aes(x=sensitivity, y=precision, color=caller) + + geom_point() + + facet_wrap(~ sample) + + labs(title="Precision recall by sample") +ggsave("roc_by_sample.pdf") + +ggplot(rocby(deldf, sizebin)) + + aes(x=sensitivity, y=precision, color=caller) + + geom_point() + + 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