More small refactorings of Mutect2 code
This commit is contained in:
David Benjamin
parent
814ad21006
commit
601c26a592
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@ -71,6 +71,7 @@ import org.broadinstitute.gatk.tools.walkers.haplotypecaller.*;
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import org.broadinstitute.gatk.tools.walkers.haplotypecaller.readthreading.ReadThreadingAssembler;
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import org.broadinstitute.gatk.utils.GenomeLoc;
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import org.broadinstitute.gatk.utils.GenomeLocParser;
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import org.broadinstitute.gatk.utils.MathUtils;
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import org.broadinstitute.gatk.utils.QualityUtils;
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import org.broadinstitute.gatk.utils.activeregion.ActiveRegion;
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import org.broadinstitute.gatk.utils.activeregion.ActiveRegionReadState;
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@ -189,20 +190,18 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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protected SampleList samplesList;
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protected boolean printTCGAsampleHeader = false;
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// fasta reference reader to supplement the edges of the reference sequence
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protected CachingIndexedFastaSequenceFile referenceReader;
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// the assembly engine
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protected LocalAssemblyEngine assemblyEngine = null;
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// the likelihoods engine
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protected ReadLikelihoodCalculationEngine likelihoodCalculationEngine = null;
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// the genotyping engine
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protected HaplotypeCallerGenotypingEngine genotypingEngine = null;
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protected SomaticGenotypingEngine genotypingEngine = null;
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private HaplotypeBAMWriter haplotypeBAMWriter;
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private byte MIN_TAIL_QUALITY;
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private double log10GlobalReadMismappingRate;
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private static final int REFERENCE_PADDING = 500;
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private static final byte MIN_TAIL_QUALITY_WITH_ERROR_CORRECTION = 6;
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private final static int maxReadsInRegionPerSample = 1000; // TODO -- should be an argument
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private final static int minReadsPerAlignmentStart = 5; // TODO -- should be an argument
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@ArgumentCollection
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protected M2ArgumentCollection MTAC = new M2ArgumentCollection();
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@ -221,10 +220,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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@Argument(fullName = "MQ_filtering_level", shortName = "MQthreshold", required = false, doc="Set an alternate MQ threshold for debugging")
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final public int MQthreshold = 20;
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public RodBinding<VariantContext> getDbsnpRodBinding() { return MTAC.dbsnp.dbsnp; }
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private HaplotypeBAMWriter haplotypeBAMWriter;
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/**
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* If a call overlaps with a record from the provided comp track, the INFO field will be annotated
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* as such in the output with the track name (e.g. -comp:FOO will have 'FOO' in the INFO field).
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@ -291,12 +286,14 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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@Argument(fullName="justDetermineActiveRegions", shortName="justDetermineActiveRegions", doc = "If specified, the HC won't actually do any assembly or calling, it'll just run the upfront active region determination code. Useful for benchmarking and scalability testing", required=false)
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protected boolean justDetermineActiveRegions = false;
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// reference base padding size
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private static final int REFERENCE_PADDING = 500;
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/**
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* As of GATK 3.3, HaplotypeCaller outputs physical (read-based) information (see version 3.3 release notes and documentation for details). This argument disables that behavior.
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*/
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@Advanced
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@Argument(fullName="doNotRunPhysicalPhasing", shortName="doNotRunPhysicalPhasing", doc="Disable physical phasing", required = false)
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protected boolean doNotRunPhysicalPhasing = false;
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private static final byte MIN_TAIL_QUALITY_WITH_ERROR_CORRECTION = 6;
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private final static int maxReadsInRegionPerSample = 1000; // TODO -- should be an argument
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private final static int minReadsPerAlignmentStart = 5; // TODO -- should be an argument
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public RodBinding<VariantContext> getDbsnpRodBinding() { return MTAC.dbsnp.dbsnp; }
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@Override
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public void initialize() {
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@ -388,7 +385,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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genotypingEngine = new SomaticGenotypingEngine( MTAC, samplesList, toolkit.getGenomeLocParser(), !doNotRunPhysicalPhasing, MTAC,
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tumorSampleName, normalSampleName, DEBUG_READ_NAME);
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genotypingEngine.setCrossHaplotypeEventMerger(variantMerger);
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genotypingEngine.setAnnotationEngine(annotationEngine);
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@ -438,7 +434,8 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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headerInfo.addAll(getM2HeaderLines());
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headerInfo.addAll(getSampleHeaderLines());
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final List<String> outputSampleNames = getOutputSampleNames();
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// if printTCGAsampleHeader, we already checked for exactly 1 tumor and 1 normal in printTCGAsampleHeader assignment in initialize()
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final List<String> outputSampleNames = printTCGAsampleHeader ? Arrays.asList("TUMOR", "NORMAL") : SampleListUtils.asList(samplesList);
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vcfWriter.writeHeader(new VCFHeader(headerInfo, outputSampleNames));
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@ -507,19 +504,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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return sampleLines;
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}
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private List<String> getOutputSampleNames(){
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if (printTCGAsampleHeader) {
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//Already checked for exactly 1 tumor and 1 normal in printTCGAsampleHeader assignment in initialize()
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final List<String> sampleNamePlaceholders = new ArrayList<>(2);
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sampleNamePlaceholders.add("TUMOR");
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sampleNamePlaceholders.add("NORMAL");
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return sampleNamePlaceholders;
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}
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else {
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return SampleListUtils.asList(samplesList);
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}
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}
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@Override
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public ActivityProfileState isActive(final RefMetaDataTracker tracker, final ReferenceContext ref, final AlignmentContext context) {
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if( context == null || context.getBasePileup().isEmpty() )
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@ -564,7 +548,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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prob = 1;
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logger.debug("At " + ref.getLocus().toString() + " tlod: " + tumorLod + " and no-normal calling");
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}
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}
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return new ActivityProfileState( ref.getLocus(), prob, ActivityProfileState.Type.NONE, null);
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@ -573,16 +556,13 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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private final static List<VariantContext> NO_CALLS = Collections.emptyList();
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@Override
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public List<VariantContext> map( final ActiveRegion originalActiveRegion, final RefMetaDataTracker metaDataTracker ) {
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if ( justDetermineActiveRegions )
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// we're benchmarking ART and/or the active region determination code in the HC, just leave without doing any work
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if ( justDetermineActiveRegions ) {
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return NO_CALLS;
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if( !originalActiveRegion.isActive() )
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// Not active so nothing to do!
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} else if( !originalActiveRegion.isActive() ) {
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return referenceModelForNoVariation(originalActiveRegion, true);
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// No reads here so nothing to do!
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if( originalActiveRegion.size() == 0 ) { return referenceModelForNoVariation(originalActiveRegion, true); }
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} else if( originalActiveRegion.size() == 0 ) {
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return referenceModelForNoVariation(originalActiveRegion, true);
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}
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logReadInfo(DEBUG_READ_NAME, originalActiveRegion.getReads(), "Present in original active region");
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// create the assembly using just high quality reads (Q20 or higher). We want to use lower
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@ -593,30 +573,25 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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assemblyActiveRegion.add(rec);
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}
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}
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logReadInfo(DEBUG_READ_NAME, assemblyActiveRegion.getReads(), "Present in assembly active region");
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// run the local assembler, getting back a collection of information on how we should proceed
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final List<VariantContext> givenAlleles = new ArrayList<>();
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final AssemblyResultSet untrimmedAssemblyResult = assembleReads(assemblyActiveRegion, givenAlleles);
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final AssemblyResultSet untrimmedAssemblyResult = assembleReads(assemblyActiveRegion, Collections.EMPTY_LIST);
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final TreeSet<VariantContext> allVariationEvents = untrimmedAssemblyResult.getVariationEvents();
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// TODO - line bellow might be unecessary : it might be that assemblyResult will always have those alleles anyway
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// TODO - so check and remove if that is the case:
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allVariationEvents.addAll(givenAlleles);
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final ActiveRegionTrimmer.Result trimmingResult = trimmer.trim(originalActiveRegion,allVariationEvents);
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// Stop the trimming madness!!!
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if (!trimmingResult.isVariationPresent())
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if (!trimmingResult.isVariationPresent()) {
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return referenceModelForNoVariation(originalActiveRegion, false);
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}
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logReadInfo(DEBUG_READ_NAME, trimmingResult.getCallableRegion().getReads(), "Present in trimming result");
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final AssemblyResultSet assemblyResult =
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trimmingResult.needsTrimming() ? untrimmedAssemblyResult.trimTo(trimmingResult.getCallableRegion()) : untrimmedAssemblyResult;
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// it is conceivable that the region is active yet has no events upon assembling only the well-mapped reads
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if( ! assemblyResult.isVariationPresent() ) {
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return referenceModelForNoVariation(originalActiveRegion, false);
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}
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final ActiveRegion regionForGenotyping = assemblyResult.getRegionForGenotyping();
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logReadInfo(DEBUG_READ_NAME, regionForGenotyping.getReads(), "Present in region for genotyping");
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@ -624,7 +599,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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haplotypeBAMWriter.addDroppedReadsFromDelta(DroppedReadsTracker.Reason.TRIMMMED, originalActiveRegion.getReads(), regionForGenotyping.getReads());
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}
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// filter out reads from genotyping which fail mapping quality based criteria
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//TODO - why don't do this before any assembly is done? Why not just once at the beginning of this method
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//TODO - on the originalActiveRegion?
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@ -639,17 +613,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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final Map<String, List<GATKSAMRecord>> perSampleFilteredReadList = splitReadsBySample(filteredReads);
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logReadInfo(DEBUG_READ_NAME, regionForGenotyping.getReads(), "Present in region for genotyping after filtering reads");
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// abort early if something is out of the acceptable range
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// TODO is this ever true at this point??? perhaps GGA. Need to check.
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if( ! assemblyResult.isVariationPresent() )
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return referenceModelForNoVariation(originalActiveRegion, false);
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// TODO is this ever true at this point??? perhaps GGA. Need to check.
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if( regionForGenotyping.size() == 0 ) {
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// no reads remain after filtering so nothing else to do!
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return referenceModelForNoVariation(originalActiveRegion, false);
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}
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// evaluate each sample's reads against all haplotypes
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final List<Haplotype> haplotypes = assemblyResult.getHaplotypeList();
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@ -658,6 +621,11 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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logReadInfo(DEBUG_READ_NAME, rec, "Present after splitting assemblyResult by sample");
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}
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//TODO: this should be written as
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//TODO final Map<String, Integer> ARreads_origNormalMQ = regionForGenotyping.getReads().stream()
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//TODO .collect(Collectors.toMap(GATKSAMRecord::getReadName, GATKSAMRecord::getMappingQuality));
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//TODO: but for some reason sometimes streamifying Mutect2 code causes a MalformedWalkerArgumentsException
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//TODO: this will probably not occur after the port to GATK4
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final HashMap<String, Integer> ARreads_origNormalMQ = new HashMap<>();
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for (final GATKSAMRecord read : regionForGenotyping.getReads()) {
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ARreads_origNormalMQ.put(read.getReadName(), read.getMappingQuality());
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@ -671,14 +639,8 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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}
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logger.debug("Computing read likelihoods with " + regionForGenotyping.getReads().size() + " reads against " + haplotypes.size() + " haplotypes across region " + assemblyResult.getRegionForGenotyping().toString());
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final ReadLikelihoods<Haplotype> readLikelihoods = likelihoodCalculationEngine.computeReadLikelihoods(assemblyResult,samplesList,reads);
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// Calculate the likelihoods: CPU intensive part.
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final ReadLikelihoods<Haplotype> readLikelihoods =
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likelihoodCalculationEngine.computeReadLikelihoods(assemblyResult,samplesList,reads);
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// Realign reads to their best haplotype.
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// KCIBUL: this is new stuff -- review it!
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final Map<GATKSAMRecord,GATKSAMRecord> readRealignments = realignReadsToTheirBestHaplotype(readLikelihoods, assemblyResult.getReferenceHaplotype(), assemblyResult.getPaddedReferenceLoc());
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if ( MTAC.bamWriter != null && MTAC.emitDroppedReads ) {
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@ -693,22 +655,14 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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logReadInfo(DEBUG_READ_NAME, rec, "Present after computing read likelihoods");
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}
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// Note: we used to subset down at this point to only the "best" haplotypes in all samples for genotyping, but there
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// was a bad interaction between that selection and the marginalization that happens over each event when computing
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// GLs. In particular, for samples that are heterozygous non-reference (B/C) the marginalization for B treats the
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// haplotype containing C as reference (and vice versa). Now this is fine if all possible haplotypes are included
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// in the genotyping, but we lose information if we select down to a few haplotypes. [EB]
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final HaplotypeCallerGenotypingEngine.CalledHaplotypes calledHaplotypes = ((SomaticGenotypingEngine)genotypingEngine).callMutations(
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haplotypes,
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final HaplotypeCallerGenotypingEngine.CalledHaplotypes calledHaplotypes = genotypingEngine.callMutations(
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readLikelihoods,
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ARreads_origNormalMQ,
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perSampleFilteredReadList,
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assemblyResult.getFullReferenceWithPadding(),
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assemblyResult.getPaddedReferenceLoc(),
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regionForGenotyping.getLocation(),
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metaDataTracker,
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givenAlleles);
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metaDataTracker);
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if ( MTAC.bamWriter != null ) {
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final Set<Haplotype> calledHaplotypeSet = new HashSet<>(calledHaplotypes.getCalledHaplotypes());
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@ -727,66 +681,7 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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}
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if( MTAC.DEBUG ) { logger.info("----------------------------------------------------------------------------------"); }
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final List<VariantContext> annotatedCalls = new ArrayList<>();
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final int eventCount = calledHaplotypes.getCalls().size();
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Integer minEventDistance = null;
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Integer maxEventDistance = null;
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Integer lastPosition = null;
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for (final VariantContext vc : calledHaplotypes.getCalls()) {
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if (lastPosition == null) {
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lastPosition = vc.getStart();
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} else {
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final int dist = Math.abs(vc.getStart() - lastPosition);
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if (maxEventDistance == null || dist > maxEventDistance) {
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maxEventDistance = dist;
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}
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if (minEventDistance == null || dist < minEventDistance) {
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minEventDistance = dist;
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}
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}
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}
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final Map<String, Object> eventDistanceAttributes = new HashMap<>();
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eventDistanceAttributes.put(GATKVCFConstants.EVENT_COUNT_IN_HAPLOTYPE_KEY, eventCount);
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eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MIN_KEY, minEventDistance);
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eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MAX_KEY, maxEventDistance);
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// can we do this with the Annotation classes instead?
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for (final VariantContext originalVC : calledHaplotypes.getCalls()) {
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final VariantContextBuilder vcb = new VariantContextBuilder(originalVC);
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final Map<String, Object> attributes = new HashMap<>(originalVC.getAttributes());
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attributes.putAll(eventDistanceAttributes);
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vcb.attributes(attributes);
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final Set<String> filters = new HashSet<>(originalVC.getFilters());
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final double tumorLod = originalVC.getAttributeAsDouble(GATKVCFConstants.TUMOR_LOD_KEY, -1);
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if (tumorLod < MTAC.TUMOR_LOD_THRESHOLD) {
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filters.add(GATKVCFConstants.TUMOR_LOD_FILTER_NAME);
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}
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// if we are in artifact detection mode, apply the thresholds for the LOD scores
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if (!MTAC.ARTIFACT_DETECTION_MODE) {
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filters.addAll(calculateFilters(metaDataTracker, originalVC, eventDistanceAttributes));
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}
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vcb.filters(filters.isEmpty() ? VariantContext.PASSES_FILTERS : filters);
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if (printTCGAsampleHeader) {
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final Genotype tumorGenotype = new GenotypeBuilder(originalVC.getGenotype(tumorSampleName)).name("TUMOR").make();
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final Genotype normalGenotype = new GenotypeBuilder(originalVC.getGenotype(normalSampleName)).name("NORMAL").make();
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vcb.genotypes(Arrays.asList(tumorGenotype, normalGenotype));
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}
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annotatedCalls.add(vcb.make());
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}
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// TODO: find a better place for this debug message
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// logger.info("We had " + TumorPowerCalculator.numCacheHits + " hits in starnd artifact power calculation");
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return annotatedCalls;
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return annotateVCs(calledHaplotypes, metaDataTracker);
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}
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private Set<String> calculateFilters(final RefMetaDataTracker metaDataTracker, final VariantContext vc, final Map<String, Object> eventDistanceAttributes) {
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@ -828,17 +723,12 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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if ( (normalAltCounts > MTAC.MAX_ALT_ALLELES_IN_NORMAL_COUNT || normalF > MTAC.MAX_ALT_ALLELE_IN_NORMAL_FRACTION ) && normalAltQualityScoreSum > MTAC.MAX_ALT_ALLELES_IN_NORMAL_QSCORE_SUM) {
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filters.add(GATKVCFConstants.ALT_ALLELE_IN_NORMAL_FILTER_NAME);
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} else {
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// NOTE: does normal alt counts presume the normal had all these events in CIS?
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if ( eventCount > 1 && normalAltCounts >= 1) {
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} else if ( eventCount > 1 && normalAltCounts >= 1) {
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filters.add(GATKVCFConstants.MULTI_EVENT_ALT_ALLELE_IN_NORMAL_FILTER_NAME);
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} else if (eventCount >= 3) {
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filters.add(GATKVCFConstants.HOMOLOGOUS_MAPPING_EVENT_FILTER_NAME);
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}
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}
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// STR contractions, that is the deletion of one repeat unit of a short repeat (>1bp repeat unit)
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// such as ACTACTACT -> ACTACT, are overwhelmingly false positives so we
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// hard filter them out by default
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@ -861,7 +751,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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filters.add(GATKVCFConstants.CLUSTERED_EVENTS_FILTER_NAME);
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}
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// clustered read position filter
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if (MTAC.ENABLE_CLUSTERED_READ_POSITION_FILTER){
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final Double tumorFwdPosMedian = (Double) vc.getAttribute(GATKVCFConstants.MEDIAN_LEFT_OFFSET_KEY);
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@ -878,7 +767,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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return filters;
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}
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private final static byte REF_MODEL_DELETION_QUAL = (byte) 30;
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/**
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* Calculate the genotype likelihoods for the sample in pileup for being hom-ref contrasted with being ref vs. alt
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@ -916,6 +804,7 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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return (normalSampleName != null);
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}
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//TODO: streamify in GATK4
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protected int getCountOfNonRefEvents(final ReadBackedPileup pileup, final byte refBase, final byte minBaseQual) {
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int i=0;
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for( final PileupElement p : pileup ) {
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@ -960,37 +849,21 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
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protected Set<GATKSAMRecord> filterNonPassingReads( final ActiveRegion activeRegion) {
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final Set<GATKSAMRecord> readsToRemove = new LinkedHashSet<>();
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for( final GATKSAMRecord rec : activeRegion.getReads() ) {
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//TODO: Takuto points out that this is questionable. Let's think hard abut it.
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// KCIBUL: only perform read quality filtering on tumor reads...
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if (isReadFromNormal(rec)) {
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if( rec.getReadLength() < MIN_READ_LENGTH ) {
|
||||
readsToRemove.add(rec);
|
||||
}
|
||||
|
||||
} else {
|
||||
|
||||
|
||||
if( rec.getReadLength() < MIN_READ_LENGTH ||
|
||||
rec.getMappingQuality() < MQthreshold ||
|
||||
BadMateFilter.hasBadMate(rec) ||
|
||||
|
||||
} else if( rec.getReadLength() < MIN_READ_LENGTH || rec.getMappingQuality() < MQthreshold || BadMateFilter.hasBadMate(rec) ||
|
||||
(keepRG != null && !rec.getReadGroup().getId().equals(keepRG)) ) {
|
||||
readsToRemove.add(rec);
|
||||
}
|
||||
}
|
||||
}
|
||||
activeRegion.removeAll(readsToRemove);
|
||||
return readsToRemove;
|
||||
}
|
||||
|
||||
private static GATKSAMRecord findReadByName(final Collection<GATKSAMRecord> reads, final String name) {
|
||||
for(final GATKSAMRecord read : reads) {
|
||||
if (name.equals(read.getReadName())) return read;
|
||||
}
|
||||
return null;
|
||||
}
|
||||
|
||||
/**
|
||||
* Instantiates the appropriate likelihood calculation engine.
|
||||
*
|
||||
|
|
@ -1025,13 +898,7 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
// enable non primary and extended reads in the active region
|
||||
@Override
|
||||
public EnumSet<ActiveRegionReadState> desiredReadStates() {
|
||||
// if ( includeUnmappedReads )
|
||||
// throw new UserException.BadArgumentValue("includeUnmappedReads", "is not yet functional");
|
||||
// else
|
||||
return EnumSet.of(
|
||||
ActiveRegionReadState.PRIMARY,
|
||||
ActiveRegionReadState.NONPRIMARY,
|
||||
ActiveRegionReadState.EXTENDED);
|
||||
return EnumSet.of(ActiveRegionReadState.PRIMARY, ActiveRegionReadState.NONPRIMARY, ActiveRegionReadState.EXTENDED);
|
||||
}
|
||||
|
||||
//---------------------------------------------------------------------------------------------------------------
|
||||
|
|
@ -1063,7 +930,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
logger.info("Ran local assembly on " + result + " active regions");
|
||||
}
|
||||
|
||||
|
||||
// The following are not used but are required by the AnnotatorCompatible interface
|
||||
public RodBinding<VariantContext> getSnpEffRodBinding() { return null; }
|
||||
public List<RodBinding<VariantContext>> getResourceRodBindings() { return Collections.emptyList(); }
|
||||
|
|
@ -1087,15 +953,12 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
final Haplotype referenceHaplotype = createReferenceHaplotype(activeRegion, paddedReferenceLoc);
|
||||
|
||||
// Create ReadErrorCorrector object if requested - will be used within assembly engine.
|
||||
ReadErrorCorrector readErrorCorrector = null;
|
||||
if (errorCorrectReads)
|
||||
readErrorCorrector = new ReadErrorCorrector(RTAC.kmerLengthForReadErrorCorrection, MIN_TAIL_QUALITY_WITH_ERROR_CORRECTION, RTAC.minObservationsForKmerToBeSolid, MTAC.DEBUG, fullReferenceWithPadding);
|
||||
|
||||
final ReadErrorCorrector readErrorCorrector = errorCorrectReads ? new ReadErrorCorrector(RTAC.kmerLengthForReadErrorCorrection, MIN_TAIL_QUALITY_WITH_ERROR_CORRECTION,
|
||||
RTAC.minObservationsForKmerToBeSolid, MTAC.DEBUG, fullReferenceWithPadding) : null;
|
||||
try {
|
||||
final AssemblyResultSet assemblyResultSet = assemblyEngine.runLocalAssembly( activeRegion, referenceHaplotype, fullReferenceWithPadding, paddedReferenceLoc, giveAlleles,readErrorCorrector );
|
||||
assemblyResultSet.debugDump(logger);
|
||||
return assemblyResultSet;
|
||||
|
||||
} catch ( final Exception e ) {
|
||||
// Capture any exception that might be thrown, and write out the assembly failure BAM if requested
|
||||
if ( captureAssemblyFailureBAM ) {
|
||||
|
|
@ -1151,10 +1014,6 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
haplotypeBAMWriter.addDroppedReadsFromDelta(DroppedReadsTracker.Reason.DOWNSAMPLED, activeRegion.getReads(), downsampledReads);
|
||||
}
|
||||
|
||||
// handle overlapping read pairs from the same fragment
|
||||
// KC: commented out as we handle overlapping read pairs in a different way...
|
||||
//cleanOverlappingReadPairs(downsampledReads, normalSampleNames);
|
||||
|
||||
activeRegion.clearReads();
|
||||
activeRegion.addAll(downsampledReads);
|
||||
activeRegion.setFinalized(true);
|
||||
|
|
@ -1189,11 +1048,7 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
|
||||
final FragmentCollection<GATKSAMRecord> fragmentCollection = FragmentUtils.create(perSampleReadList);
|
||||
for ( final List<GATKSAMRecord> overlappingPair : fragmentCollection.getOverlappingPairs() )
|
||||
|
||||
// in MuTect -- right now we compare the
|
||||
FragmentUtils.adjustQualsOfOverlappingPairedFragments(overlappingPair);
|
||||
|
||||
|
||||
}
|
||||
}
|
||||
|
||||
|
|
@ -1236,20 +1091,61 @@ public class MuTect2 extends ActiveRegionWalker<List<VariantContext>, Integer> i
|
|||
|
||||
private boolean isReadFromNormal(final GATKSAMRecord rec) {
|
||||
return normalSampleName != null && normalSampleName.equals(rec.getReadGroup().getSample());
|
||||
|
||||
}
|
||||
|
||||
public String getTumorSampleName(){
|
||||
return tumorSampleName;
|
||||
}
|
||||
|
||||
// KCIBUL: new stuff -- read up on this!!
|
||||
/**
|
||||
* As of GATK 3.3, HaplotypeCaller outputs physical (read-based) information (see version 3.3 release notes and documentation for details). This argument disables that behavior.
|
||||
*/
|
||||
@Advanced
|
||||
@Argument(fullName="doNotRunPhysicalPhasing", shortName="doNotRunPhysicalPhasing", doc="Disable physical phasing", required = false)
|
||||
protected boolean doNotRunPhysicalPhasing = false;
|
||||
final List<VariantContext> annotateVCs(final HaplotypeCallerGenotypingEngine.CalledHaplotypes calledHaplotypes, final RefMetaDataTracker metaDataTracker) {
|
||||
final int eventCount = calledHaplotypes.getCalls().size();
|
||||
final Map<String, Object> eventDistanceAttributes = new HashMap<>(); //TODO: should be Map<String, Integer> -- see TODO below
|
||||
eventDistanceAttributes.put(GATKVCFConstants.EVENT_COUNT_IN_HAPLOTYPE_KEY, eventCount);
|
||||
if (eventCount > 1) {
|
||||
final int lastPosition = calledHaplotypes.getCalls().get(0).getStart();
|
||||
final int[] eventDistances = new int[calledHaplotypes.getCalls().size() - 1];
|
||||
for (int n = 0; n < eventDistances.length; n++) {
|
||||
eventDistances[n] = Math.abs(calledHaplotypes.getCalls().get(n + 1).getStart() - lastPosition);
|
||||
}
|
||||
eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MIN_KEY, MathUtils.arrayMin(eventDistances));
|
||||
eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MAX_KEY, MathUtils.arrayMax(eventDistances));
|
||||
} else { //TODO: putting null is a hack -- we should remove this and update the integration test md5s
|
||||
eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MIN_KEY, null);
|
||||
eventDistanceAttributes.put(GATKVCFConstants.EVENT_DISTANCE_MAX_KEY, null);
|
||||
}
|
||||
|
||||
final List<VariantContext> annotatedCalls = new ArrayList<>();
|
||||
// can we do this with the Annotation classes instead?
|
||||
for (final VariantContext originalVC : calledHaplotypes.getCalls()) {
|
||||
final VariantContextBuilder vcb = new VariantContextBuilder(originalVC);
|
||||
|
||||
final Map<String, Object> attributes = new HashMap<>(originalVC.getAttributes());
|
||||
attributes.putAll(eventDistanceAttributes);
|
||||
vcb.attributes(attributes);
|
||||
|
||||
final Set<String> filters = new HashSet<>(originalVC.getFilters());
|
||||
|
||||
final double tumorLod = originalVC.getAttributeAsDouble(GATKVCFConstants.TUMOR_LOD_KEY, -1);
|
||||
if (tumorLod < MTAC.TUMOR_LOD_THRESHOLD) {
|
||||
filters.add(GATKVCFConstants.TUMOR_LOD_FILTER_NAME);
|
||||
}
|
||||
|
||||
// if we are in artifact detection mode, apply the thresholds for the LOD scores
|
||||
if (!MTAC.ARTIFACT_DETECTION_MODE) {
|
||||
filters.addAll(calculateFilters(metaDataTracker, originalVC, eventDistanceAttributes));
|
||||
}
|
||||
|
||||
vcb.filters(filters.isEmpty() ? VariantContext.PASSES_FILTERS : filters);
|
||||
|
||||
if (printTCGAsampleHeader) {
|
||||
final Genotype tumorGenotype = new GenotypeBuilder(originalVC.getGenotype(tumorSampleName)).name("TUMOR").make();
|
||||
final Genotype normalGenotype = new GenotypeBuilder(originalVC.getGenotype(normalSampleName)).name("NORMAL").make();
|
||||
vcb.genotypes(Arrays.asList(tumorGenotype, normalGenotype));
|
||||
}
|
||||
annotatedCalls.add(vcb.make());
|
||||
}
|
||||
return annotatedCalls;
|
||||
}
|
||||
}
|
||||
|
||||
|
||||
|
|
|
|||
|
|
@ -51,10 +51,7 @@
|
|||
|
||||
package org.broadinstitute.gatk.tools.walkers.cancer.m2;
|
||||
|
||||
import com.google.java.contract.Ensures;
|
||||
import htsjdk.samtools.util.StringUtil;
|
||||
import htsjdk.variant.variantcontext.*;
|
||||
import org.apache.commons.collections.ListUtils;
|
||||
import org.apache.commons.lang.mutable.MutableDouble;
|
||||
import org.apache.commons.lang.mutable.MutableInt;
|
||||
import org.apache.log4j.Logger;
|
||||
|
|
@ -63,7 +60,6 @@ import org.broadinstitute.gatk.tools.walkers.genotyper.afcalc.AFCalculatorProvid
|
|||
import org.broadinstitute.gatk.tools.walkers.haplotypecaller.HaplotypeCallerGenotypingEngine;
|
||||
import org.broadinstitute.gatk.utils.GenomeLoc;
|
||||
import org.broadinstitute.gatk.utils.GenomeLocParser;
|
||||
import org.broadinstitute.gatk.utils.commandline.RodBinding;
|
||||
import org.broadinstitute.gatk.utils.contexts.ReferenceContext;
|
||||
import org.broadinstitute.gatk.utils.genotyper.MostLikelyAllele;
|
||||
import org.broadinstitute.gatk.utils.genotyper.PerReadAlleleLikelihoodMap;
|
||||
|
|
@ -87,6 +83,9 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
private final String matchedNormalSampleName;
|
||||
private final String DEBUG_READ_NAME;
|
||||
|
||||
//Mutect2 does not run in GGA mode
|
||||
private static final List<VariantContext> NO_GIVEN_ALLELES = Collections.EMPTY_LIST;
|
||||
|
||||
// {@link GenotypingEngine} requires a non-null {@link AFCalculatorProvider} but this class doesn't need it. Thus we make a dummy
|
||||
private static AFCalculatorProvider DUMMY_AF_CALCULATOR_PROVIDER = new AFCalculatorProvider() {
|
||||
public AFCalculator getInstance(final int ploidy, final int maximumAltAlleles) { return null; }
|
||||
|
|
@ -109,6 +108,7 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
this.DEBUG_READ_NAME = DEBUG_READ_NAME;
|
||||
|
||||
// coverage related initialization
|
||||
//TODO: in GATK4, use a QualityUtils method
|
||||
final double errorProbability = Math.pow(10, -MTAC.POWER_CONSTANT_QSCORE/10);
|
||||
strandArtifactPowerCalculator = new TumorPowerCalculator(errorProbability, MTAC.STRAND_ARTIFACT_LOD_THRESHOLD, 0.0f);
|
||||
}
|
||||
|
|
@ -118,48 +118,40 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
* genotype likelihoods and assemble into a list of variant contexts and genomic events ready for calling
|
||||
*
|
||||
* The list of samples we're working with is obtained from the readLikelihoods
|
||||
|
||||
* @param haplotypes Haplotypes to assign likelihoods to
|
||||
* @param readLikelihoods Map from reads->(haplotypes,likelihoods)
|
||||
* @param perSampleFilteredReadList Map from sample to reads that were filtered after assembly and before calculating per-read likelihoods.
|
||||
* @param ref Reference bytes at active region
|
||||
* @param refLoc Corresponding active region genome location
|
||||
* @param activeRegionWindow Active window
|
||||
* @param activeAllelesToGenotype Alleles to genotype
|
||||
*
|
||||
* @return A CalledHaplotypes object containing a list of VC's with genotyped events and called haplotypes
|
||||
*
|
||||
*/
|
||||
// TODO - can this be refactored? this is hard to follow!
|
||||
public CalledHaplotypes callMutations (
|
||||
final List<Haplotype> haplotypes,
|
||||
final ReadLikelihoods<Haplotype> readLikelihoods,
|
||||
final Map<String, Integer> originalNormalReadQualities,
|
||||
final Map<String, List<GATKSAMRecord>> perSampleFilteredReadList,
|
||||
final byte[] ref,
|
||||
final GenomeLoc refLoc,
|
||||
final GenomeLoc activeRegionWindow,
|
||||
final RefMetaDataTracker tracker,
|
||||
final List<VariantContext> activeAllelesToGenotype) {
|
||||
// sanity check input arguments
|
||||
if (haplotypes == null || haplotypes.isEmpty()) throw new IllegalArgumentException("haplotypes input should be non-empty and non-null, got "+haplotypes);
|
||||
final RefMetaDataTracker tracker) {
|
||||
//TODO: in GATK4 use Utils.nonNull
|
||||
if (readLikelihoods == null || readLikelihoods.sampleCount() == 0) throw new IllegalArgumentException("readLikelihoods input should be non-empty and non-null, got "+readLikelihoods);
|
||||
if (ref == null || ref.length == 0 ) throw new IllegalArgumentException("ref bytes input should be non-empty and non-null, got "+ref);
|
||||
if (refLoc == null || refLoc.size() != ref.length) throw new IllegalArgumentException(" refLoc must be non-null and length must match ref bytes, got "+refLoc);
|
||||
if (activeRegionWindow == null ) throw new IllegalArgumentException("activeRegionWindow must be non-null, got "+activeRegionWindow);
|
||||
if (activeAllelesToGenotype == null ) throw new IllegalArgumentException("activeAllelesToGenotype must be non-null, got "+activeAllelesToGenotype);
|
||||
|
||||
final List<Haplotype> haplotypes = readLikelihoods.alleles();
|
||||
|
||||
// Somatic Tumor/Normal Sample Handling
|
||||
if (!readLikelihoods.samples().contains(tumorSampleName)) {
|
||||
throw new IllegalArgumentException("readLikelihoods does not contain the tumor sample " + tumorSampleName);
|
||||
}
|
||||
|
||||
// if we don't have the normal sample, we are in tumor only mode
|
||||
// TODO: check in MuTect2.java for code we can skip when in tumor only mode
|
||||
final boolean hasNormal = matchedNormalSampleName != null;
|
||||
|
||||
// update the haplotypes so we're ready to call, getting the ordered list of positions on the reference
|
||||
// that carry events among the haplotypes
|
||||
final TreeSet<Integer> startPosKeySet = decomposeHaplotypesIntoVariantContexts(haplotypes, readLikelihoods, ref, refLoc, activeAllelesToGenotype);
|
||||
final TreeSet<Integer> startPosKeySet = decomposeHaplotypesIntoVariantContexts(haplotypes, readLikelihoods, ref, refLoc, NO_GIVEN_ALLELES);
|
||||
|
||||
// Walk along each position in the key set and create each event to be outputted
|
||||
final Set<Haplotype> calledHaplotypes = new HashSet<>();
|
||||
|
|
@ -170,7 +162,7 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
continue;
|
||||
}
|
||||
|
||||
final List<VariantContext> eventsAtThisLoc = getVCsAtThisLocation(haplotypes, loc, activeAllelesToGenotype);
|
||||
final List<VariantContext> eventsAtThisLoc = getVCsAtThisLocation(haplotypes, loc, NO_GIVEN_ALLELES);
|
||||
|
||||
if( eventsAtThisLoc.isEmpty() ) { continue; }
|
||||
|
||||
|
|
@ -203,14 +195,12 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
* If we just want a map of Alleles to Haplotypes, we should be able to do so directly; no need for intermediate maps, which just complicates the code.
|
||||
**/
|
||||
|
||||
|
||||
final Map<Allele, List<Haplotype>> alleleMapper = createAlleleMapper(mergeMap, eventMapper);
|
||||
|
||||
// converting ReadLikelihoods<Haplotype> to ReadLikeliHoods<Allele>
|
||||
ReadLikelihoods<Allele> readAlleleLikelihoods = readLikelihoods.marginalize(alleleMapper, genomeLocParser.createPaddedGenomeLoc(genomeLocParser.createGenomeLoc(mergedVC), ALLELE_EXTENSION));
|
||||
|
||||
//LDG: do we want to do this before or after pulling out overlapping reads?
|
||||
// TODO: do we want this at all? How does downsampling help?
|
||||
if (MTAC.isSampleContaminationPresent()) {
|
||||
readAlleleLikelihoods.contaminationDownsampling(MTAC.getSampleContamination());
|
||||
}
|
||||
|
|
@ -232,24 +222,18 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
tumorLods.set(altAllele, tumorHetGenotypeLLs.get(altAllele) - tumorHetGenotypeLLs.getRef());
|
||||
}
|
||||
|
||||
|
||||
// TODO: another good breakpoint e.g. compute normal LOD/set thresholds
|
||||
// TODO: anything related to normal should be encapsulated in Optional
|
||||
|
||||
// Normal LOD must exceed this threshold for the variant to make it in the vcf
|
||||
// TODO: variable name too log
|
||||
double normalLodThresholdForVCF = -Double.MIN_VALUE;
|
||||
|
||||
// A variant candidate whose normal LOD is below this threshold will be filtered as 'germline_risk'
|
||||
// This is a more stringent threshold than normalLodThresholdForVCF
|
||||
double normalLodFilterThreshold = -Double.MIN_VALUE;
|
||||
|
||||
double normalLodFilterThreshold = -Double.MAX_VALUE;
|
||||
PerReadAlleleLikelihoodMap normalPRALM = null;
|
||||
final PerAlleleCollection<Double> normalLods = PerAlleleCollection.createPerAltAlleleCollection();
|
||||
|
||||
// if normal bam is available, compute normal LOD
|
||||
// TODO: this if statement should be a standalone method for computing normal LOD
|
||||
// TODO: then we can do something like normalLodThreshold = hasNormal ? thisMethod() : Optional.empty()
|
||||
// if normal bam is available, compute normal LOD
|
||||
if (hasNormal) {
|
||||
normalPRALM = readAlleleLikelihoods.toPerReadAlleleLikelihoodMap(readAlleleLikelihoods.sampleIndex(matchedNormalSampleName));
|
||||
filterPRALMForOverlappingReads(normalPRALM, mergedVC.getReference(), loc, true);
|
||||
|
|
@ -258,10 +242,8 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
final GenomeLoc eventGenomeLoc = genomeLocParser.createGenomeLoc(activeRegionWindow.getContig(), loc);
|
||||
final Collection<VariantContext> cosmicVC = tracker.getValues(MTAC.cosmicRod, eventGenomeLoc);
|
||||
final Collection<VariantContext> dbsnpVC = tracker.getValues(MTAC.dbsnp.dbsnp, eventGenomeLoc);
|
||||
|
||||
final boolean germlineAtRisk = !dbsnpVC.isEmpty() && cosmicVC.isEmpty();
|
||||
|
||||
normalLodThresholdForVCF = MTAC.INITIAL_NORMAL_LOD_THRESHOLD;
|
||||
normalLodFilterThreshold = germlineAtRisk ? MTAC.NORMAL_DBSNP_LOD_THRESHOLD : MTAC.NORMAL_LOD_THRESHOLD;
|
||||
|
||||
// compute normal LOD = LL(X|REF)/LL(X|ALT) where REF is the diploid HET with AF = 0.5
|
||||
|
|
@ -284,7 +266,7 @@ public class SomaticGenotypingEngine extends HaplotypeCallerGenotypingEngine {
|
|||
|
||||
for (final Allele altAllele : mergedVC.getAlternateAlleles()) {
|
||||
final boolean passesTumorLodThreshold = tumorLods.getAlt(altAllele) >= MTAC.INITIAL_TUMOR_LOD_THRESHOLD;
|
||||
final boolean passesNormalLodThreshold = hasNormal ? normalLods.getAlt(altAllele) >= normalLodThresholdForVCF : true;
|
||||
final boolean passesNormalLodThreshold = hasNormal ? normalLods.getAlt(altAllele) >= MTAC.INITIAL_NORMAL_LOD_THRESHOLD : true;
|
||||
if (passesTumorLodThreshold && passesNormalLodThreshold) {
|
||||
numPassingAlts++;
|
||||
allelesThatPassThreshold.add(altAllele);
|
||||
|
|
|
|||
|
|
@ -70,8 +70,6 @@ public class MuTect2IntegrationTest extends WalkerTest {
|
|||
final static String DREAM3_TP_INTERVALS_FILE = privateTestDir + "m2_dream3.tp.intervals";
|
||||
final static String DREAM3_FP_INTERVALS_FILE = privateTestDir + "m2_dream3.fp.intervals";
|
||||
|
||||
|
||||
|
||||
final String commandLine =
|
||||
"-T MuTect2 --no_cmdline_in_header -dt NONE --disableDithering -alwaysloadVectorHMM -pairHMM LOGLESS_CACHING -ip 50 -R %s --dbsnp %s --cosmic %s --normal_panel %s -I:tumor %s -I:normal %s -L %s";
|
||||
|
||||
|
|
@ -160,9 +158,6 @@ public class MuTect2IntegrationTest extends WalkerTest {
|
|||
M2Test(CCLE_MICRO_TUMOR_BAM, CCLE_MICRO_NORMAL_BAM, CCLE_MICRO_INTERVALS_FILE, "-contamination 0.1", "c25e48edd704bbb436cd6456d9f47d8b");
|
||||
}
|
||||
|
||||
/**
|
||||
* Test that tumor-only mode does not create an empty vcf
|
||||
*/
|
||||
@Test
|
||||
public void testTumorOnly(){
|
||||
m2TumorOnlyTest(CCLE_MICRO_TUMOR_BAM, "2:166000000-167000000", "", "2af2253b1f09ea8fd354e1bf2c4612f0");
|
||||
|
|
@ -177,6 +172,4 @@ public class MuTect2IntegrationTest extends WalkerTest {
|
|||
public void testClusteredReadPositionFilter() {
|
||||
M2Test(DREAM3_TUMOR_BAM, DREAM3_NORMAL_BAM, DREAM3_FP_INTERVALS_FILE, "--enable_clustered_read_position_filter", "b44c23af7de84f96d2371db25d29aba2");
|
||||
}
|
||||
|
||||
|
||||
}
|
||||
|
|
|
|||
Loading…
Reference in New Issue