{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T03:07:50Z","timestamp":1785208070637,"version":"3.55.0"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T00:00:00Z","timestamp":1768176000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82170527"],"award-info":[{"award-number":["82170527"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007219","name":"Natural Science Foundation of Shanghai Municipality","doi-asserted-by":"publisher","award":["21ZR1452600"],"award-info":[{"award-number":["21ZR1452600"]}],"id":[{"id":"10.13039\/100007219","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Genet."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Non-syndromic biliary atresia (BA) is a multifactorial disorder with a poorly understood genetic basis. We previously identified 154 BA-associated SNPs spanning the\n                      <jats:italic>ADD3<\/jats:italic>\n                      locus, which harbors the most strongly associated common variants in Asian populations. The susceptibility SNPs are in high linkage disequilibrium, but the causal variants remain unidentified.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Using available databases, we predicted the regulatory potential of the 154 BA-associated SNPs. To functionally validate these findings, we assessed the enhancer activity of\n                      <jats:italic>cis<\/jats:italic>\n                      -regulatory elements (CREs) containing the risk variants using a dual-luciferase reporter assay. The role of\n                      <jats:italic>ADD3<\/jats:italic>\n                      dysregulation in hepatobiliary development was investigated using zebrafish. The spatiotemporal expression pattern of the\n                      <jats:italic>ADD3<\/jats:italic>\n                      ortholog in zebrafish embryos was mapped by\n                      <jats:italic>in situ<\/jats:italic>\n                      hybridization. Additionally, we performed mRNA overexpression and morpholino knockdown to examine the effects of perturbing\n                      <jats:italic>ADD3<\/jats:italic>\n                      ortholog expression on zebrafish hepatobiliary development.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Among 154 associated SNPs, 28 clustered within 10 putative CREs with predicted enhancer function.\n                      <jats:italic>In vitro<\/jats:italic>\n                      allele-specific luciferase assays demonstrated enhancer activity in eight of these CREs, with risk haplotypes at three loci driving significantly higher activity than non-risk haplotypes (\n                      <jats:italic>P &amp;lt;<\/jats:italic>\n                      0.05). The zebrafish\n                      <jats:italic>add3a<\/jats:italic>\n                      gene, an ortholog of human\n                      <jats:italic>ADD3<\/jats:italic>\n                      , was expressed in developing livers. Both overexpression and knockdown of\n                      <jats:italic>add3a<\/jats:italic>\n                      in zebrafish disrupted hepatobiliary function and development, resulting in gallbladder hypoplasia\/agenesis and reduced intrahepatic bile duct density. These phenotypes closely recapitulate BA pathology observed in humans. Combined with our prior data linking risk alleles to heightened\n                      <jats:italic>ADD3<\/jats:italic>\n                      expression and demonstrating\n                      <jats:italic>ADD3<\/jats:italic>\n                      overexpression in BA livers, these results indicate that genetic variants drive ADD3 upregulation, thereby predisposing to BA development.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Multiple risk variants within enhancers upregulated\n                      <jats:italic>ADD3<\/jats:italic>\n                      expression, which contributed to BA pathogenesis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fgene.2025.1733215","type":"journal-article","created":{"date-parts":[[2026,1,11]],"date-time":"2026-01-11T22:10:19Z","timestamp":1768169419000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Multiple susceptibility enhancer variants increasing ADD3 expression predisposes to biliary atresia risk"],"prefix":"10.3389","volume":"16","author":[{"given":"Xinru","family":"Han","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoyue","family":"Pei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meirong","family":"Bai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xun","family":"Chu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,12]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1002\/hep.32107","article-title":"Biliary organoids uncover delayed epithelial development and barrier function in biliary atresia","volume":"75","author":"Amarachintha","year":"2022","journal-title":"Hepatology"},{"key":"B2","doi-asserted-by":"publisher","first-page":"342","DOI":"10.1038\/nrgastro.2015.74","article-title":"Pathogenesis of biliary atresia: defining biology to understand clinical phenotypes","volume":"12","author":"Asai","year":"2015","journal-title":"Nat. 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