{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,4,26]],"date-time":"2026-04-26T08:32:54Z","timestamp":1777192374539,"version":"3.51.4"},"posted":{"date-parts":[[2024,11,18]]},"group-title":"Genetic and Genomic Medicine","reference-count":35,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T00:00:00Z","timestamp":1731888000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Nonsyndromic cleft lip (nsCL) exhibits a non-random laterality pattern, with left-sided clefts occurring approximately twice as frequently as right-sided clefts. The molecular mechanisms underlying this laterality bias remain poorly understood.<\/jats:p>\n                <jats:p>We performed whole-genome sequencing and methylation profiling on a family comprising monozygotic twins with mirror-image nsCL, their affected mother, and unaffected father and brother. We conducted three independent replications via (1) publicly available whole genome data; (2) genome-wide methylation analysis in 38 individuals with unilateral cleft; and (3) validation of methylation results in the top 3 candidate genes in 385 unrelated individuals with unilateral clefts (DNA from blood or saliva).<\/jats:p>\n                <jats:p>\n                  We identified a variant in\n                  <jats:italic>FGF20<\/jats:italic>\n                  (p.Ile79Val) shared by the twins and their mother. We observed laterality and severity-associated methylation differences in three main genes.\n                  <jats:italic>ARID5B<\/jats:italic>\n                  showed higher methylation in left clefts (saliva, p=0.001; blood, p=0.032).\n                  <jats:italic>ZFP57<\/jats:italic>\n                  demonstrated a strong cleft-extent effect, with cleft lip and palate (CLP) showing markedly higher methylation than cleft lip only (CL) (LCLP vs. RCL p\n                  <jats:sup>adj<\/jats:sup>\n                  =0.0004; LCLP vs. LCL p\n                  <jats:sup>adj<\/jats:sup>\n                  = 0.019).\n                  <jats:italic>HOOK2<\/jats:italic>\n                  displayed a cross-tissue cleft-extent effect in the opposite direction \u2014 CLP subtypes were hypomethylated relative to CL-only subtypes in blood (p&lt;0.0001) and saliva p=0.0008).\n                <\/jats:p>\n                <jats:p>\n                  This study provides evidence that DNA methylation patterns plays a role in both the laterality and severity of cleft lip.\n                  <jats:italic>ARID5B<\/jats:italic>\n                  provides a consistent laterality signal across tissues, while\n                  <jats:italic>ZFP57<\/jats:italic>\n                  and\n                  <jats:italic>HOOK2<\/jats:italic>\n                  track palatal involvement independently of side. Together, these findings suggest that epigenetic variation acts downstream of genetic predisposition to shape cleft phenotypes.\n                <\/jats:p>","DOI":"10.1101\/2024.11.16.24317351","type":"posted-content","created":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T22:05:14Z","timestamp":1731967514000},"source":"Crossref","is-referenced-by-count":0,"title":["Genetic and Epigenetic Contributors to Cleft Laterality: Evidence from Monozygotic Mirror Twins and Replication Cohorts"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2239-8120","authenticated-orcid":false,"given":"Aline L.","family":"Petrin","sequence":"first","affiliation":[{"name":"Department of Orthodontics, College of Dentistry and Dental Clinics, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5223-6846","authenticated-orcid":false,"given":"Waheed","family":"Awotoye","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, College of Dentistry and Dental Clinics, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina E.","family":"Spencer","sequence":"additional","affiliation":[{"name":"College of Dentistry and Dental Clinics, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4730-1866","authenticated-orcid":false,"given":"Ligiane A.","family":"Machado-Paula","sequence":"additional","affiliation":[{"name":"Iowa Institute of Human Genetics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3945-2309","authenticated-orcid":false,"given":"Luke","family":"Hovey","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, Tufts University School of Dental Medicine, Boston, MA, 02111, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3548-7839","authenticated-orcid":false,"given":"Henry","family":"Keen","sequence":"additional","affiliation":[{"name":"Iowa Institute of Human Genetics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4433-9976","authenticated-orcid":false,"given":"Michael","family":"Chimenti","sequence":"additional","affiliation":[{"name":"Iowa Institute of Human Genetics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, 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USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James C.","family":"Thomas","sequence":"additional","affiliation":[{"name":"Division of Biostatistics and Computational Biology, College of Dentistry and Dental Clinics, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1229-5964","authenticated-orcid":false,"given":"Azeez","family":"Butali","sequence":"additional","affiliation":[{"name":"Department of Oral Pathology, Radiology, & Medicine, College of Dentistry and Dental Clinics, University of Iowa, Iowa City, IA, 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6346-6551","authenticated-orcid":false,"given":"Jeffrey C.","family":"Murray","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, 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