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[BIOIN-1570] Fixed edge case in variant annotation #8810

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Jun 3, 2024
Merged

[BIOIN-1570] Fixed edge case in variant annotation #8810

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1e21c8c
[BIOIN-1570] Fixed edge case in variant annotation when the variant i…
ilyasoifer May 3, 2024
4466f85
[BIOIN-1570] Fixed comments
ilyasoifer May 3, 2024
500460e
logger -> OneShotLogger
ilyasoifer May 29, 2024
9cc86d8
PR comments
ilyasoifer May 31, 2024
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50 changes: 40 additions & 10 deletions ...in/java/org/broadinstitute/hellbender/tools/walkers/annotator/flow/FlowAnnotatorBase.java
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Original file line number Diff line number Diff line change
Expand Up @@ -34,7 +34,6 @@
* are accumulated as well.
*/
public abstract class FlowAnnotatorBase implements InfoFieldAnnotation {
private final static Logger logger = LogManager.getLogger(FlowAnnotatorBase.class);
protected final OneShotLogger flowMissingOneShotLogger = new OneShotLogger(FlowAnnotatorBase.class);


Expand Down Expand Up @@ -202,7 +201,7 @@ protected void variantType(final VariantContext vc, final LocalContext localCont
if (isSpecial(alleles.get(i))){
continue;
}
if ((localContext.hmerIndelLength.get(i)==null) || (localContext.hmerIndelLength.get(i)==0)){
if ((localContext.hmerIndelLength==null) || (localContext.hmerIndelLength.get(i)==null) || (localContext.hmerIndelLength.get(i)==0)){
isHmer=false;
}
}
Expand Down Expand Up @@ -250,7 +249,12 @@ protected void isHmerIndel(final VariantContext vc, final LocalContext localCont
// get byte before and after
final byte before = getReferenceNucleotide(localContext, vc.getStart() - 1);
final byte[] after = getReferenceHmerPlus(localContext, vc.getEnd() + 1, MOTIF_SIZE);

if (after.length==0){
flowMissingOneShotLogger.warn("Failed to get hmer from reference, probably because the " +
"variant is very close to the end of the chromosome, isHmerIndel and RightMotif annotations will not be calculated. " +
"Variant position: " + vc.getContig() + ":" + vc.getEnd() + 1);
return;
}
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I'm looking inside of establishReadGroupFlowOrder() and it looks like it tries to disable annotation if it can't establish what the flow order is for the local context (which is explicitly the case for the user that brought up this issue)... Can you disentangle what is supposed to happen for this annotation class in the event that no-flow order can be established? The annotation requires a flow order in order to determine what kind of indel it is no?

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there is a method called isActualFlowOrderRequired in this abstract class that returns false, the annotations that the user enabled all inherit from that abstract class and do not require flow order. The only annotation that does require it is CycleSkipStatus.

// build two haplotypes. add byte before and after
final byte[] refHap = buildHaplotype(before, ref.getBases(), after);
final byte[] altHap = buildHaplotype(before, alt.getBases(), after);
Expand Down Expand Up @@ -338,6 +342,12 @@ protected void getLeftMotif(final VariantContext vc, final LocalContext localCon
}

String motif = getRefMotif(localContext, vc.getStart() - MOTIF_SIZE, MOTIF_SIZE);
if (motif.length() != MOTIF_SIZE){
flowMissingOneShotLogger.warn("Failed to get motif from reference, probably because the variant is very close to the " +
"start of the chromosome. LeftMotif annotation will not be calculated. " +
"Variant position: " + vc.getContig() + ":" + vc.getStart());
return;
}
if ( a.length() != refLength ) {
motif = motif.substring(1) + vc.getReference().getBaseString().substring(0, 1);
}
Expand All @@ -350,8 +360,13 @@ protected void getLeftMotif(final VariantContext vc, final LocalContext localCon
protected void getRightMotif(final VariantContext vc, final LocalContext localContext) {

final int refLength = vc.getReference().length();
final String motif = getRefMotif(localContext, vc.getStart() + refLength, MOTIF_SIZE);

String motif = getRefMotif(localContext, vc.getStart() + refLength, MOTIF_SIZE);
if (motif.length() != MOTIF_SIZE){
flowMissingOneShotLogger.warn("Failed to get motif from reference, probably because " +
"the variant is close to the end of the chromosome. RightMotif annotation will not be calculated. " +
"Variant position: " + vc.getContig() + ":" + vc.getStart());
return;
}
// fill empty entries (non indel alelles)
for ( int i = 0 ; i < localContext.rightMotif.size() ; i++ ) {
if ( localContext.rightMotif.get(i) == null ) {
Expand All @@ -366,6 +381,11 @@ protected void gcContent(final VariantContext vc, final LocalContext localContex

final int begin = vc.getStart() - (GC_CONTENT_SIZE / 2);
final String seq = getRefMotif(localContext, begin + 1, GC_CONTENT_SIZE);
if ( seq.length() != GC_CONTENT_SIZE ) {
flowMissingOneShotLogger.warn("gcContent will not be calculated at position " + vc.getContig() + ":" + vc.getStart() +
" because the variant is too close to the edge of the chromosome");
return;
}
int gcCount = 0;
for ( byte b : seq.getBytes() ) {
if ( b == 'G' || b == 'C' ) {
Expand Down Expand Up @@ -424,19 +444,25 @@ protected void cycleSkip(final VariantContext vc, final LocalContext localContex
localContext.attributes.put(GATKVCFConstants.FLOW_CYCLESKIP_STATUS, css);
}

// get a single nucleoid from reference
// get a single nucleotid from reference
private byte getReferenceNucleotide(final LocalContext localContext, final int start) {
final int index = start - localContext.ref.getWindow().getStart();
final byte[] bases = localContext.ref.getBases();
Utils.validIndex(index, bases.length);
Utils.validIndex(index, bases.length); // do not catch, if here the location is outside of the reference, there is a problem!
return bases[index];
}

// get an hmer from reference plus a number of additional bases
private byte[] getReferenceHmerPlus(final LocalContext localContext, final int start, final int additional) {
int index = start - localContext.ref.getWindow().getStart();
final byte[] bases = localContext.ref.getBases();
Utils.validIndex(index, bases.length);
try {
Utils.validIndex(index, bases.length);
} catch (IllegalArgumentException e) {
flowMissingOneShotLogger.warn("Failed to get hmer from reference, too close to the edge. " +
"Position: " + localContext.ref.getContig() + ":" + index);
return new byte[0];
}

// get hmer
final StringBuilder sb = new StringBuilder();
Expand All @@ -458,8 +484,12 @@ private String getRefMotif(final LocalContext localContext, final int start, fin
final byte[] bases = localContext.ref.getBases();
final int startIndex = start - localContext.ref.getWindow().getStart();
final int endIndex = startIndex + length;
Utils.validIndex(startIndex, bases.length);
Utils.validIndex(endIndex-1, bases.length);
try {
Utils.validIndex(startIndex, bases.length);
Utils.validIndex(endIndex-1, bases.length);
} catch (IllegalArgumentException e) {
return "";
}
return new String(Arrays.copyOfRange(bases, startIndex, endIndex));
}

Expand Down
21 changes: 21 additions & 0 deletions ...ava/org/broadinstitute/hellbender/tools/walkers/annotator/flow/FlowAnnotatorUnitTest.java
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Expand Up @@ -69,7 +69,28 @@ public Object[][] getTestData() {
// ins hmer indel
"TATCT C ATTGACCAA", "CA",
"ins", "1", "2", "A", "ATCTC", "TTGAC", "0.3", "NA", "h-indel"
},
//The following tests test the edge case of variant so close to the edge of the chromosome
//that some of the annotations can't be calculated
{
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Can you add to these comments a little more context for the nature of this error and what those are testing?

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Added comments

// Close to 5' edge of the chromosome, expect not to crash on finding the left motif
"ATCT C ATTGACCAA", "CA",
"ins", "1", "2", "A", null, "TTGAC", "0.3", "NA", "h-indel"
},
{
// close to 3' edge of the chromosome, should not find left motif or GC content
"TATCT C ATT", "CA",
"ins", "1", "2", "A", "ATCTC", null, null, "NA", "h-indel"
},
{
// very close to 3' edge of the chromosome, should not find left motif, GC content or if it is
// an hmer indel
"TATCT C ", "CA",
"ins", "1", null, null, "ATCTC", null, null, "NA", "non-h-indel"
}



};

return testData;
Expand Down
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