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

Renato Assis Machado; Lilianny Querino Rocha de Oliveira; Ana Lúcia Carrinho Ayroza Rangel; Silvia Regina de Almeida Reis; Rafaela Scariot; Daniella Reis Barbosa Martelli; Hercílio Martelli-Júnior; Ricardo D Coletta

doi:10.3390/dj11010007

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

Dent J (Basel). 2022 Dec 26;11(1):7. doi: 10.3390/dj11010007.

Affiliations

¹ Department of Oral Diagnosis, School of Dentistry, University of Campinas, Piracicaba 13414-018, São Paulo, Brazil.
² Hospital for Rehabilitation of Craniofacial Anomalies, University of São Paulo, Bauru 17012-900, São Paulo, Brazil.
³ Graduate Program in Oral Biology, School of Dentistry, University of Campinas, Piracicaba 13414-018, São Paulo, Brazil.
⁴ Center of Biological Sciences and of the Health, School of Dentistry, State University of Western Paraná, Cascavel 85819-110, Paraná, Brazil.
⁵ Department of Basic Science, Bahiana School of Medicine and Public Health, Salvador 40290-000, Bahia, Brazil.
⁶ Department of Oral and Maxillofacial Surgery, School of Health Science, Federal University of Paraná, Curitiba 80060-000, Parana, Brazil.
⁷ Stomatology Clinic, Dental School, State University of Montes Claros, Montes Claros 39401-089, Minas Gerais, Brazil.
⁸ Center for Rehabilitation of Craniofacial Anomalies, Dental School, University of Professor Edson Antônio Velano, Alfenas 37130-000, Minas Gerais, Brazil.

Abstract

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±P) 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±P 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 ≤0.01. 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±P reached only nominal p values, NSCLO risk was significantly higher in carriers of the FOS rs1046117 C allele (OR: 1.28, 95% CI: 1.10-1.64, p = 0.004), TC heterozygous genotype (OR: 1.59, 95% CI: 1.16-2.18, p = 0.003), and in the dominant model (OR: 1.50, 95% CI: 1.10-2.02, 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±P, 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.

Keywords: cleft lip; cleft lip and palate; genetic epistasis; single-nucleotide polymorphism.

Grants and funding

NA/Fundação de Amparo à Pesquisa do Estado de Minas Gerais