5ad55adbb6a5cc72a393700130584aa87fef2c89
lrnassar
  Tue Jun 30 06:15:44 2026 -0700
varFreqs: add Top 3 source AFs to mouseOvers; audit excludes SGDP and SVatalog. refs #36642

Adds a Top 3 source AFs ranking to the varFreqsAffected and varFreqsBackground
mouseOvers. Alongside the pooled allele frequency, the mouseOver now lists the
three cohorts/arms with the highest per-source AF, formatted as
"Source (AF), Source (AF), Source (AF)". Disease cohorts with phenotype splits
carry the arm label (SPARK ASD, SCHEMA case, GREGoR unaffected); population
cohorts use the bare key. Per-population sub-ancestries are deliberately
excluded so a high sub-pop AF cannot crowd out actual project-level signals.

vcfToBigBed.py adds a top_n_source_afs helper, collects per-arm AFs into
affected_arm_afs / background_arm_afs, and emits two new fields
topAffectedSources and topBackgroundSources. AS schema field count 163 -> 165.

An AF-distribution sweep across all 28 source cohorts identified SGDP and
SVatalog as encoding allele counts per genotyped individual (small N, AF
defaults near 0.5), making their per-source AF unreliable for the ranking.
Adds a skip_top_ranking column (col 9) to databases.tsv, set to 1 for SGDP
and SVatalog, and gates the per-arm AF append in vcfToBigBed.py on this
flag. Both cohorts still contribute to pooled backgroundAC/AN/AF and still
appear in backgroundSources; they are only suppressed from the Top 3.

Description pages varFreqsAffected.html and varFreqsBackground.html document
the ranking; the latter also documents the SGDP/SVatalog exclusion. Build
documentation in varFreqs.txt is updated.

diff --git src/hg/makeDb/trackDb/human/varFreqsAffected.html src/hg/makeDb/trackDb/human/varFreqsAffected.html
index e1cd9bbefab..bb67d401445 100644
--- src/hg/makeDb/trackDb/human/varFreqsAffected.html
+++ src/hg/makeDb/trackDb/human/varFreqsAffected.html
@@ -1,149 +1,164 @@
 <h2>Description</h2>
 <p>
 This track shows small variants (SNVs and short indels) that were observed in
 <b>affected or case individuals</b> of disease-study cohorts, annotated with their
 predicted protein consequence and colored by severity. It is one half of a matched pair:
 the companion
 <a href="hgTrackUi?g=varFreqsBackground">Population reference</a> track shows the same
 kind of variants seen in population reference cohorts and in unaffected relatives or
 controls. Displaying the two together lets you compare, for example, how often a
 loss-of-function variant in a gene of interest is seen in affected individuals versus the
 general/unaffected background. For the full list of contributing projects, see the
 <a href="hgTrackUi?g=varFreqs">SNV Frequencies</a> collection page.
 </p>
 <p>
 The affected counts are drawn from the affected or case arm of five disease-study cohorts:
 SFARI SPARK WES and SFARI SPARK WGS (autism spectrum disorder probands), SCHEMA
 (schizophrenia cases), GREGoR (affected rare-disease participants), and GA4K (a pediatric
 rare-disease cohort). For SPARK, SFARI WGS, SCHEMA, and GREGoR, the source data carries an
 explicit affected/unaffected (or case/control) label, and only the affected arm feeds this
 track. GA4K reports a single cohort-wide frequency with no per-individual label; because it
 is a rare-disease cohort, it is counted as affected here, with the caveat that it enrolls
 parent-child trios, so a minority of its carriers are unaffected parents. Genotyping-array
 cohorts are not included in either track.
 </p>
 
 <h2>Display Conventions</h2>
 <h3>Color by Consequence</h3>
 <p>Variants are colored by their most severe predicted consequence:</p>
 <table class="stdTbl">
 <tr><th>Color</th><th>Consequence class</th><th>Examples</th></tr>
 <tr><th style="background-color:#FF0000;width:2em">&nbsp;</th>
     <td>Protein-truncating / loss-of-function</td>
     <td>stop_gained, frameshift, splice_donor, splice_acceptor, stop_lost, start_lost</td></tr>
 <tr><th style="background-color:#1F77B4;width:2em">&nbsp;</th>
     <td>Missense / in-frame</td>
     <td>missense, inframe_insertion, inframe_deletion, protein_altering</td></tr>
 <tr><th style="background-color:#008000;width:2em">&nbsp;</th>
     <td>Synonymous</td>
     <td>synonymous, stop_retained</td></tr>
 <tr><th style="background-color:#808080;width:2em">&nbsp;</th>
     <td>Non-coding / intergenic</td>
     <td>intron, non_coding, intergenic, UTR</td></tr>
 </table>
 <p>
 The score (used for shading) is the pooled affected/case allele frequency times 1000.
 </p>
 
 <h3>Pooled allele frequency</h3>
 <p>
 <b>Affected AF</b> is the pooled rate across contributing affected arms:
 <code>affectedAF = sum(AC) / sum(AN)</code>, where <b>affectedAC</b> sums the allele counts
 and <b>affectedAN</b> sums the allele numbers across each cohort/arm that provides both AC and
 AF (the per-arm AN is derived as <code>round(AC / AF)</code>). Cohorts that publish only AF
 (with no AC or AN of their own) are still pooled by assigning them an assumed allele number,
 set as a <code>default_an</code> in the build configuration; their per-arm AC is then derived
 as <code>round(AF &times; default_an)</code>. Cohorts
 that publish only AC and have no <code>default_an</code> set (currently GREGoR's per-arm
 AC_AFFECTED/UNAFFECTED/UNKNOWN) are listed in <b>affectedCohorts</b> but do not contribute
 to the pool numerator or denominator; their carriers are visible in the per-database AC
 column instead. The pooled rate is preferred over a max-across-cohorts statistic so a
 small cohort with a high local AF cannot dominate the displayed frequency.
 </p>
 
+<h3>Top affected sources by AF</h3>
+<p>
+Alongside the pooled rate, the mouseover lists the top 3 contributing
+affected arms ranked by their own per-source AF, formatted as
+<code>Source (AF)</code>. This surfaces case cohorts where the variant is
+specifically enriched, even when the pooled rate across all arms is small.
+For disease cohorts that ship a phenotype split (SPARK, SFARI WGS, SCHEMA,
+GREGoR), the displayed AF is the affected-arm AF and the label includes the
+arm (for example <code>SPARK ASD</code>, <code>SCHEMA case</code>); for
+cohorts with no split (GA4K) the label is just the cohort name and the AF
+is the whole-cohort AF. Arms that ship only AC and no AF (currently GREGoR
+per-arm) are not included in this ranking because no AF is available;
+they still appear in <b>affectedCohorts</b>.
+</p>
+
 <h3>Finding case-enriched loss-of-function variants</h3>
 <p>
 To look for protein-truncating variants that are common in affected individuals but rare
 in the background, set the Consequence filter to Stop Gained, Frameshift, Splice Donor and
 Splice Acceptor (these appear red), then add an upper limit on the
 <b>Background AF</b> filter. Each variant here carries both its affected frequency and its
 background frequency, so this isolates variants seen in cases with little or no presence in
 the population/unaffected set. Comparing visually against the
 <a href="hgTrackUi?g=varFreqsBackground">Population reference</a> track shows the same
 contrast across a whole gene.
 </p>
 
 <h2>Filters</h2>
 <ul>
   <li><b>Variant Type</b> and <b>Consequence</b>: restrict to SNV/insertion/deletion/MNV
       and to predicted consequence classes (the Consequence filter uses OR logic over the
       comma-separated tokens on each variant).</li>
   <li><b>Affected/case AF</b>, <b>AC</b>, <b>AN</b>: the pooled allele frequency
       (sum AC / sum AN), summed allele count, and summed allele number across the
       contributing affected arms. See &quot;Pooled allele frequency&quot; above.</li>
   <li><b>Background AF</b>, <b>AC</b>, <b>AN</b>: the same triple computed across the
       population + unaffected background, for filtering case-enriched variants.</li>
   <li><b>Affected/case cohort</b>: restrict to variants seen in specific disease cohorts
       (for example, only the two autism cohorts).</li>
   <li><b>Reference/Alternate Length</b> and <b>Length Change</b>: filter by allele length.</li>
 </ul>
 
 <h2>Methods</h2>
 <p>
 Variant-frequency VCFs from the contributing cohorts were stripped of unneeded INFO fields,
 normalized with <code>bcftools norm</code> (splitting multi-allelic sites), and merged with
 <code>bcftools merge</code>. The merged callset was annotated with predicted protein
 consequences using <a href="https://samtools.github.io/bcftools/howtos/csq-calling.html"
 target="_blank">bcftools csq</a> against the
 <a href="https://www.ensembl.org/info/data/ftp/index.html" target="_blank">Ensembl</a>
 GRCh38 release 115 gene models.
 </p>
 <p>
 A custom Python script (<code>vcfToBigBed.py</code>) then read the per-cohort allele
 counts and frequencies and, for each variant, pooled the allele counts and allele numbers
 across the affected arms (case/proband subgroups, plus GA4K whole-cohort) to produce this
 track, and across the population cohorts and unaffected/control subgroups to produce the
 companion <a href="hgTrackUi?g=varFreqsBackground">Population reference</a> track. A variant
 seen in both groups appears in both tracks. The build is documented in the
 <a href="https://github.com/ucscGenomeBrowser/kent/blob/master/src/hg/makeDb/doc/hg38/varFreqs.txt"
 target="_blank">makeDoc</a>, and the scripts are on
 <a href="https://github.com/ucscGenomeBrowser/kent/tree/master/src/hg/makeDb/scripts/varFreqs"
 target="_blank">GitHub</a>.
 </p>
 
 <h2>Data Access</h2>
 <p>
 Because the merged callset combines cohorts whose redistribution licenses differ, this
 track is <b>not available for download</b> and is not in the Table Browser. It can be
 reconstructed from the individual source VCFs using the
 <a href="https://github.com/ucscGenomeBrowser/kent/tree/master/src/hg/makeDb/scripts/varFreqs"
 target="_blank">conversion scripts</a> and the
 <a href="https://github.com/ucscGenomeBrowser/kent/blob/master/src/hg/makeDb/doc/hg38/varFreqs.txt"
 target="_blank">build documentation</a>. The per-project subtracks on the
 <a href="hgTrackUi?g=varFreqs">SNV Frequencies</a> collection page document how to obtain
 each source dataset.
 </p>
 
 <h2>Credits</h2>
 <p>
 This track is only possible thanks to the data from the participants and families of the
 SFARI SPARK, SCHEMA, GREGoR and GA4K studies. Click the individual project subtracks on the
 <a href="hgTrackUi?g=varFreqs">SNV Frequencies</a> collection page for the specific credits
 and citations of each cohort. Thanks to Alex Ioannidis, UCSC, for the inspiration for this
 track and to Andreas Lahner, MGZ, for feedback.
 </p>
 
 <h2>References</h2>
 <p>
 For the primary citation of each source cohort, see the References section on the
 <a href="hgTrackUi?g=varFreqs">SNV Frequencies</a> collection page. The merged-track build
 uses the following tools:
 </p>
 <p>
 Danecek P, McCarthy SA.
 <a href="https://doi.org/10.1093/bioinformatics/btx100" target="_blank">
 BCFtools/csq: haplotype-aware variant consequences</a>.
 <em>Bioinformatics</em>. 2017 Jul 1;33(13):2037-2039.
 PMID: <a href="https://www.ncbi.nlm.nih.gov/pubmed/28205675" target="_blank">28205675</a>;
 PMC: <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5870570/" target="_blank">PMC5870570</a>
 </p>