{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T20:29:50Z","timestamp":1776371390457,"version":"3.51.2"},"reference-count":36,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,12,26]],"date-time":"2022-12-26T00:00:00Z","timestamp":1672012800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais (FAPEMIG)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Dentistry Journal"],"abstract":"<jats:p>Associations of CRISPLD2 (cysteine-rich secretory protein LCCL domain containing 2) and genes belonging to its activation pathway, including FOS (Fos proto-oncogene), CASP8 (caspase 8) and MMP2 (matrix metalloproteinase 2), with nonsyndromic orofacial cleft risk, have been reported, but the results are yet unclear. The aim of this study was to evaluate single nucleotide polymorphisms (SNPs) in FOS, CASP8 and MMP2 and to determine their SNP-SNP interactions with CRISPLD2 variants in the risk of nonsyndromic cleft lip with or without cleft palate (NSCL\u00b1P) in the Brazilian population. The SNPs rs1046117 (FOS), rs3769825 (CASP8) and rs243836 (MMP2) were genotyped using TaqMan allelic discrimination assays in a case-control sample containing 801 NSCL\u00b1P patients (233 nonsyndromic cleft lip only (NSCLO) and 568 nonsyndromic cleft lip and palate (NSCLP)) and 881 healthy controls via logistic regression analysis adjusted for the effects of sex and genomic ancestry proportions with a multiple comparison p value set at \u22640.01. SNP-SNP interactions with rs1546124, rs8061351, rs2326398 and rs4783099 in CRISPLD2 were performed with the model-based multifactor dimensionality reduction test complemented with a 1000 permutation-based strategy. Although the association between FOS rs1046117 and risk of NSCL\u00b1P reached only nominal p values, NSCLO risk was significantly higher in carriers of the FOS rs1046117 C allele (OR: 1.28, 95% CI: 1.10\u20131.64, p = 0.004), TC heterozygous genotype (OR: 1.59, 95% CI: 1.16\u20132.18, p = 0.003), and in the dominant model (OR: 1.50, 95% CI: 1.10\u20132.02, p = 0.007). Individually, no significant associations between cleft risk and the SNPs in CASP8 and MMP2 were observed. SNP-SNP interactions involving CRISPLD2 variants and rs1046117 (FOS), rs3769825 (CASP8) and rs243836 (MMP2) yielded several significant p values, mostly driven by FOS rs1046117 and CASP8 rs3769825 in NSCL\u00b1P, FOS rs1046117 in NSCLO and CRISPLD2 rs8061351 in NSCLP. Our study is the first in the Brazilian population to reveal the association of FOS rs1046117 with NSCLO risk, and to support that CRISPLD2, CASP8, FOS and MMP2 interactions may be related to the pathogenesis of this common craniofacial malformation.<\/jats:p>","DOI":"10.3390\/dj11010007","type":"journal-article","created":{"date-parts":[[2022,12,27]],"date-time":"2022-12-27T07:30:06Z","timestamp":1672126206000},"page":"7","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Brazilian Multiethnic Association Study of Genetic Variant Interactions among FOS, CASP8, MMP2 and CRISPLD2 in the Risk of Nonsyndromic Cleft Lip with or without Cleft Palate"],"prefix":"10.3390","volume":"11","author":[{"given":"Renato Assis","family":"Machado","sequence":"first","affiliation":[{"name":"Department of Oral Diagnosis, School of Dentistry, University of Campinas, Piracicaba 13414-018, S\u00e3o Paulo, Brazil"},{"name":"Hospital for Rehabilitation of Craniofacial Anomalies, University of S\u00e3o Paulo, Bauru 17012-900, S\u00e3o Paulo, Brazil"},{"name":"Graduate Program in Oral Biology, School of Dentistry, University of Campinas, Piracicaba 13414-018, S\u00e3o Paulo, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lilianny Querino Rocha","family":"de Oliveira","sequence":"additional","affiliation":[{"name":"Graduate Program in Oral Biology, School of Dentistry, University of Campinas, Piracicaba 13414-018, S\u00e3o Paulo, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana L\u00facia Carrinho Ayroza","family":"Rangel","sequence":"additional","affiliation":[{"name":"Center of Biological Sciences and of the Health, School of Dentistry, State University of Western Paran\u00e1, Cascavel 85819-110, Paran\u00e1, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvia Regina de Almeida","family":"Reis","sequence":"additional","affiliation":[{"name":"Department of Basic Science, Bahiana School of Medicine and Public Health, Salvador 40290-000, Bahia, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rafaela","family":"Scariot","sequence":"additional","affiliation":[{"name":"Department of Oral and Maxillofacial Surgery, School of Health Science, Federal University of Paran\u00e1, Curitiba 80060-000, Parana, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniella Reis Barbosa","family":"Martelli","sequence":"additional","affiliation":[{"name":"Stomatology Clinic, Dental School, State University of Montes Claros, Montes Claros 39401-089, Minas Gerais, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Herc\u00edlio","family":"Martelli-J\u00fanior","sequence":"additional","affiliation":[{"name":"Stomatology Clinic, Dental School, State University of Montes Claros, Montes Claros 39401-089, Minas Gerais, Brazil"},{"name":"Center for Rehabilitation of Craniofacial Anomalies, Dental School, University of Professor Edson Ant\u00f4nio Velano, Alfenas 37130-000, Minas Gerais, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5285-3046","authenticated-orcid":false,"given":"Ricardo D.","family":"Coletta","sequence":"additional","affiliation":[{"name":"Department of Oral Diagnosis, School of Dentistry, University of Campinas, Piracicaba 13414-018, S\u00e3o Paulo, Brazil"},{"name":"Graduate Program in Oral Biology, School of Dentistry, University of Campinas, Piracicaba 13414-018, S\u00e3o Paulo, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1038\/nrg2933","article-title":"Cleft lip and palate: Understanding genetic and environmental influences","volume":"12","author":"Dixon","year":"2011","journal-title":"Nat. Rev. Genet."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1016\/S0140-6736(09)60695-4","article-title":"Cleft lip and palate","volume":"374","author":"Mossey","year":"2009","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2800","DOI":"10.12688\/f1000research.9503.1","article-title":"Genetic factors influencing risk to orofacial clefts: Today\u2019s challenges and tomorrow\u2019s opportunities","volume":"5","author":"Beaty","year":"2016","journal-title":"F1000Research"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1590\/S1806-83242007000400006","article-title":"Prevalence of nonsyndromic oral clefts in a reference hospital in the state of Minas Gerais, Brazil, between 2000\u20132005","volume":"21","author":"Porto","year":"2007","journal-title":"Braz. Oral Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1590\/S1806-83242009000100007","article-title":"Prevalence of orofacial clefts and social factors in Brazil","volume":"23","author":"Rodrigues","year":"2009","journal-title":"Braz. Oral Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1111\/odi.14146","article-title":"Genetic and epigenetic studies in non-syndromic oral clefts","volume":"28","author":"Alade","year":"2022","journal-title":"Oral Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2241","DOI":"10.1093\/hmg\/ddm176","article-title":"CRISPLD2: A novel NSCLP candidate gene","volume":"16","author":"Chiquet","year":"2007","journal-title":"Hum. Mol. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1002\/bdra.20840","article-title":"The CRISPLD2 gene is involved in cleft lip and\/or cleft palate in a Chinese population","volume":"91","author":"Shen","year":"2011","journal-title":"Birth Defects Res. A Clin. Mol. Teratol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1177\/1055665617738995","article-title":"Association Between CRISPLD2 Polymorphisms and the Risk of Nonsyndromic Clefts of the Lip and\/or Palate: A Meta-analysis","volume":"55","author":"Ge","year":"2018","journal-title":"Cleft Palate Craniofac. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1002\/dvg.22051","article-title":"Craniofacial abnormalities result from knock down of nonsyndromic clefting gene, crispld2, in zebrafish","volume":"50","author":"Yuan","year":"2012","journal-title":"Genesis"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1441","DOI":"10.1038\/s41431-018-0192-5","article-title":"Knockdown of Crispld2 in zebrafish identifies a novel network for nonsyndromic cleft lip with or without cleft palate candidate genes","volume":"26","author":"Chiquet","year":"2018","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"826","DOI":"10.1177\/154405910208101206","article-title":"Matrix metalloproteinases have a role in palatogenesis","volume":"81","author":"Brown","year":"2002","journal-title":"J. Dent. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1111\/jop.12470","article-title":"Brazilian multicenter study of association between polymorphisms in CRISPLD2 and JARID2 and non-syndromic oral clefts","volume":"46","author":"Messetti","year":"2017","journal-title":"J. Oral Pathol. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1002\/ajmg.a.35830","article-title":"Polymorphisms at regions 1p22.1 (rs560426) and 8q24 (rs1530300) are risk markers for nonsyndromic cleft lip and\/or palate in the Brazilian population","volume":"161","author":"Bagordakis","year":"2013","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1002\/bdra.23204","article-title":"Analysis of susceptibility polymorphisms for nonsyndromic cleft lip with or without cleft palate in the Brazilian population","volume":"100","author":"Messetti","year":"2014","journal-title":"Birth Defects Res. A Clin. Mol. Teratol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2344","DOI":"10.1002\/ajmg.a.37181","article-title":"Genetic risk factors for nonsyndromic cleft lip with or without cleft palate in a Brazilian population with high African ancestry","volume":"167","author":"Hoshi","year":"2015","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1590\/S0103-64402007000200012","article-title":"A simple and cost-effective protocol for DNA isolation from buccal epithelial cells","volume":"18","author":"Aidar","year":"2007","journal-title":"Braz. Dent. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1093\/genetics\/155.2.945","article-title":"Inference of population structure using multilocus genotype data","volume":"155","author":"Pritchard","year":"2000","journal-title":"Genetics"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2198","DOI":"10.1093\/bioinformatics\/btq352","article-title":"mbmdr: An R package for exploring gene-gene interactions associated with binary or quantitative traits","volume":"26","author":"Calle","year":"2010","journal-title":"Bioinformatics"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1111\/j.1600-0722.2010.00743.x","article-title":"CRISPLD2 polymorphisms are associated with non-syndromic cleft lip with or without cleft palate in a northern Chinese population","volume":"118","author":"Shi","year":"2010","journal-title":"Eur. J. Oral Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1245","DOI":"10.1177\/0022034516657003","article-title":"Association Studies and Direct DNA Sequencing Implicate Genetic Susceptibility Loci in the Etiology of Nonsyndromic Orofacial Clefts in Sub-Saharan African Populations","volume":"95","author":"Gowans","year":"2016","journal-title":"J. Dent. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1055\/s-0040-1713166","article-title":"CRISPLD2 Gene Polymorphisms with Nonsyndromic Cleft Lip Palate in Indian Population","volume":"7","author":"Neela","year":"2020","journal-title":"Glob. Med. Genet."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1111\/j.1600-0722.2010.00801.x","article-title":"No evidence for a role of CRISPLD2 in non-syndromic cleft lip with or without cleft palate in an Italian population","volume":"119","author":"Girardi","year":"2011","journal-title":"Eur. J. Oral Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.mod.2018.05.006","article-title":"Fos metamorphoses: Lessons from mutants in model organisms","volume":"154","year":"2018","journal-title":"Mech. Dev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1186\/s13046-020-01686-9","article-title":"Selective antagonism of cJun for cancer therapy","volume":"39","author":"Brennan","year":"2020","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/nprot.2015.123","article-title":"SIFT missense predictions for genomes","volume":"11","author":"Vaser","year":"2016","journal-title":"Nat. Protoc."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"D886","DOI":"10.1093\/nar\/gky1016","article-title":"CADD: Predicting the deleteriousness of variants throughout the human genome","volume":"47","author":"Rentzsch","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1093\/hmg\/ddm060","article-title":"Loss of MMP-2 disrupts skeletal and craniofacial development and results in decreased bone mineralization, joint erosion and defects in osteoblast and osteoclast growth","volume":"16","author":"Mosig","year":"2007","journal-title":"Hum. Mol. Genet."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1091\/mbc.12.5.1457","article-title":"TGF-beta3-induced palatogenesis requires matrix metalloproteinases","volume":"12","author":"Blavier","year":"2001","journal-title":"Mol. Biol. Cell"},{"key":"ref_30","first-page":"129","article-title":"Apoptosis and MMP-2, TIMP-2 expression in cleft lip and palate","volume":"15","author":"Smane","year":"2013","journal-title":"Stomatologija"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1042\/bj20020707","article-title":"A functional activating protein 1 (AP-1) site regulates matrix metalloproteinase 2 (MMP-2) transcription by cardiac cells through interactions with JunB-Fra1 and JunB-FosB heterodimers","volume":"369","author":"Bergman","year":"2003","journal-title":"Biochem. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1002\/bdra.23026","article-title":"Association of MMP3 and TIMP2 promoter polymorphisms with nonsyndromic oral clefts","volume":"94","author":"Letra","year":"2012","journal-title":"Birth Defects Res. A Clin. Mol. Teratol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1558","DOI":"10.1002\/bdr2.1768","article-title":"Cellular and developmental basis of orofacial clefts","volume":"112","author":"Ji","year":"2020","journal-title":"Birth Defects Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1306","DOI":"10.1111\/odi.14174","article-title":"Revisiting the embryogenesis of lip and palate development","volume":"28","author":"Hammond","year":"2022","journal-title":"Oral Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1177\/0022034511408613","article-title":"Tgf-beta-mediated FasL-Fas-Caspase pathway is crucial during palatogenesis","volume":"90","author":"Huang","year":"2011","journal-title":"J. Dent. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"L391","DOI":"10.1152\/ajplung.00119.2014","article-title":"LGL1 modulates proliferation, apoptosis, and migration of human fetal lung fibroblasts","volume":"308","author":"Zhang","year":"2015","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."}],"container-title":["Dentistry Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-6767\/11\/1\/7\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:51:52Z","timestamp":1760147512000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-6767\/11\/1\/7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,26]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["dj11010007"],"URL":"https:\/\/doi.org\/10.3390\/dj11010007","relation":{},"ISSN":["2304-6767"],"issn-type":[{"value":"2304-6767","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,26]]}}}