EPIDEMIOLOGICAL STUDY OF CLEFT PALATE IN THE STATE OF BAHIA, BRAZIL

IntroductionNon-syndromic cleft lip with or without cleft palate is a common worldwide birth defect due to a combination of environmental and genetic factors. Genome-wide association studies reported the rs7078160 of Vax1 is closely related to non-syndromic cleft lip with or without cleft palate in European populations. The following studies showed the same results in Mongolian, Japanese, Filipino, Vietnamese populations etc. However, conflicting research had been reported in Chinese population, ObjectiveThe aim of this study was to investigate the association between the rs7078160 polymorphism and non-syndromic cleft lip with or without cleft palate in Southern Chinese patients. MethodsIn this study, we investigated the polymorphism distribution of rs7078160 in 100 complete patient trios (39 patients with non-syndromic cleft lip and palate; 36 patients with non-syndromic cleft lip only; 25 had non-syndromic cleft palate only; and their parents) from Southern ethnic Han Chinese. 60 healthy trios were selected as control. Polymerase chain reaction and Sanger sequencing were used to genotype rs7078160 in Vax1; both case–control and family-based associations were analyzed. ResultsThe case–control analyses revealed the rs7078160 polymorphism was significant, associated with non-syndromic cleft lip with or without cleft palate (p=0.04) and non-syndromic cleft lip and palate (p=0.01), but not associated with non-syndromic cleft lip only and non-syndromic cleft palate only patients. The genotype composition of rs7078160 comprises mutated homozygous AA, heterozygous AG and wild homozygous GG. Cases with AG+AA genotypes compared with GG homozygotes showed an increased risk of non-syndromic cleft lip with or without cleft palate (p=0.04, OR=2.05, 95% CI: 1.01–4.16) and non-syndromic cleft lip and palate (p=0.01, OR=3.94, 95% CI: 1.34–11.54). In addition, we did not detect any transmission-disequilibrium in rs7078160 (p=0.68). ConclusionThis study suggests that rs7078160 polymorphism is a risk factor of non-syndromic cleft lip with or without cleft palate, and Vax1 is strongly associated with non-syndromic cleft lip with or without cleft palate in Southern Chinese Han populations.

Background Cleft lip with or without cleft palate (CL/P) is the most common facial birth defect, with a worldwide incidence of 1 in 700-1000 live births. CL/P can be divided into syndromic CL/P (SCL/P) and nonsyndromic CL/P (NSCL/P). Genetic factors are an important component to the etiology of NSCL/P. ARHGAP29, one of the NSCL/P disease-causing genes, mediates the cyclical regulation of small GTP binding proteins such as RhoA and plays an essential role in cellular shape, proliferation, and craniofacial development. Methods The present study investigated a Chinese family with NSCL/P and explored potential pathogenic variants using whole-exome sequencing (WES). Variants were screened and filtered through bioinformatic analysis and prediction of variant pathogenicity. Cosegregation was subsequently conducted. Results We identified a novel heterozygous missense variant of ARHGAP29 (c.2615C > T, p.A872V) in a Chinese pedigree with NSCL/P. Conclusion We detected the disease-causing variant in this NSCL/P family. Our identification expands the genetic spectrum of ARHGAP29 and contributes to novel approaches to the genetic diagnosis and counseling of CL/P families.

PURPOSE: Nonsyndromic cleft lip and/or palate (NSCLP) is the most common congenital craniofacial anomaly. Early recognition of any associated developmental delay is critical to counseling families and developing individualized treatment plans. Here we sought to identify developmental delays associated with NSCLP in a large population of children in order to begin identifying etiology and improve multidisciplinary management. METHODS: This is an IRB-approved, single-center retrospective analysis of all patients with a diagnosis of cleft lip and/or cleft palate between 5 and 21 years of age. Demographic and clinical variables were collected from this patient population as well as from children comprising the 2018 National Survey of Children’s Health database. RESULTS: All children with an identified or suspected genetic syndrome were excluded (160 in our cohort and 1383 in the National Survey of Children’s Health database). Subsequently, 619 children in our cohort and 29,147 in the National Survey of Children’s Health database were identified with NSCLP and included in our analysis. The mean birth weight amongst NSCLP children was lower than that in the national cohort (108.5 ± 24.8 oz versus 117.8 ± 19.1 oz; P < 0.0001). Nearly one-fourth (25.8%) of children with NSCLP were admitted to the NICU at birth. The distribution of cleft lip/palate diagnoses in the NSCLP cohort is shown. Compared with the national cohort, children with isolated cleft palate had significantly higher rates of intellectual disability (3.2% versus 0.5%, P < 0.00001), speech delay (70.8% versus 7.1%, P < 0.00001), global developmental delay (15.7% versus 5.8%, P < 0.00001), cerebral palsy (2.2% versus 0.3%, P < 0.00001), and hearing loss (25.9% versus 1.0%, P < 0.00001). Rates of learning disability (7.0% versus 5.9%, P = 0.529), behavioral delay (7.6% versus 11.4%, P = 0.1038), ADD/ADHD (2.7% versus 2.3%, P = 0.7032), autism (4.3% versus 5.5%, P = 0.5005), and vision loss (1.6% versus 1.2%, P = 0.5764) were comparable between those with isolated cleft palate and the national cohort. Children with cleft lip (with or without cleft palate) had significantly higher rates of ADD/ADHD compared with the normative national cohort: isolated cleft lip (7.7% versus 2.3%, P = 0.0092), unilateral cleft lip and palate (4.6% versus 2.3%, P = 0.0088), bilateral cleft lip & palate (5.9% versus 2.3%, P = 0.0153). CONCLUSIONS: Our study demonstrates, for the first time, higher rates of various developmental delays in children with NSCLP compared with the general pediatric population. This includes increased rates of intellectual disability, global delay, and cerebral palsy in children with nonsyndromic isolated cleft palate and increased ADD/ADHD in children with cleft lip (with or without cleft palate). The association of NSCLP diagnoses with developmental delays highlights the importance of proper risk assessment of patients, appropriate family counseling, and multi-disciplinary team management.

Nonsyndromic cleft lip and/or cleft palate (NSCLP) are common congenital anomalies in humans, the etiologies of which are complex and associated with both genetic and environmental factors. Previous data suggested single nucleotide polymorphisms (SNPs) of rs1546124, rs4783099, and rs16974880 of the CRISPLD2 gene were associated with an increased risk of NSCLP; however, subsequent studies have yielded conflicting results. This study aims to evaluate the associations of the aforementioned polymorphisms with NSCLP in a Northwestern Chinese population. Three CRISPLD2 SNPs were genotyped in a case-control study (n = 907), including 444 NSCLP patients and 463 healthy individuals, using polymerase chain reaction-denaturing high-performance liquid chromatography (PCR-DHPLC). The genotype and allele frequencies of rs1546124 (odds ratio [OR], 2.30; 95% confidence interval [CI], 1.58-3.34; p = 1 × 10(-5) ) and rs4783099 (OR, 0.73; 95% CI, 0.54-1.00; p = 0.05) were different in NSCLP patients compared with controls. Furthermore, the CC genotype at rs1546124 was associated with increased risk for cleft lip with or without cleft palate (CL/P; OR, 2.11; 95% CI, 1.41-3.15; p(correct) = 1.5 × 10(-4) ) and for cleft palate only (CPO; OR, 2.93; 95% CI, 1.69-5.07; p(correct) = 5.4 × 10(-4) ), whereas the T allele of rs4783099 was associated with decreased risk for CPO. Further gender stratification showed that the statistical association of these two loci is mainly in the male patients, and not in female patients. Our results suggest that the CRISPLD2 gene contributes to the etiology of NSCLP in the Northwestern Chinese population. SNP rs1546124 is significantly related to NSCLP, associated with both CL/P and CPO groups, and SNP rs4783099 is significantly associated with CPO.

Association between FOXE1 and non-syndromic orofacial clefts in a northeastern Chinese population

The nonsyndromic cleft lip and/or palate (NSCL/P) is a common congenital orofacial defect of multifactorial origin with involvement of genetic and nutritional factors. The presence of genetic polymorphisms in enzymes that metabolize folic acid and vitamin supplements used to prevent these anomalies have not yet been well established in the literature. The aim of this study was to conduct a review of the literature about the role of folate and gene polymorphisms in the metabolic pathway of folic acid as regards the occurrence of NSCL/P. We reviewed theoretical material that addressed data on these topics. According to the studies analyzed in the literature, there was no consensus with regard to the protective effect of supplemental folic acid taken in the periconceptional period in the prevention of NSCL/P. The studies that evaluated defects in genes involved in folate metabolism, found an association of some polymorphisms with NSCL/P in different populations.

Hypermethylation of WNT3A gene and non-syndromic cleft lip and/or palate in association with in utero exposure to lead: A mediation analysis

To analyse the sex ratio of non-syndromic cleft lip and/or palate (NSCL/P) in Hong Kong Chinese. All cases of NSCL/P delivered in a teaching hospital in Hong Kong from 2001–2014 were reviewed. The sex ratio for different kinds of NSCL/P was compared using chi square test. During the study period, 87,989 (45,562 male and 42,427 female) live births were delivered. There were 96 cases of NSCL/P, 50 male and 46 female. The incidence of NSCL/P was 1:916 live births (1:911 male and 1:922 female). There were 54 (56%) cleft lip and palate, 24 (25%) cleft lip only and 18 (19%) cleft palate only. Male was more frequently affected than female, with male: female (M:F) ratio of 1.1:1. This feature of male predominance was observed in cleft lip only, cleft lip and palate and cleft lip with or without cleft palate, with M:F ratio of 1.7:1, 1.3:1 and 1.4:1 respectively. However, the sex ratio was reversed in cleft palate only, with M:F ratio of 0.3:1. The difference is statistically significant (p = 0.0175). Non-syndromic cleft lip with or without palate is more common in male while non-syndromic cleft palate only is more common in female. This may indicate that they represent two different entities with totally different genetic basis and environmental trigger. Researchers working on NSCL/P may need to make a distinction between the two entities.

Comprehensive analysis of plasma miRNA and related ceRNA network in non-syndromic cleft lip and/or palate.

Background:Nonsyndromic cleft lip and/or palate (NSCLP) is the most common congenital craniofacial anomaly. Early recognition of developmental delays associated with NSCLP is critical for counseling and management. This study investigates developmental delays in a large population of children with NSCLP.Methods:This is an institutional review board–approved, retrospective analysis of children 5–21 years of age with a diagnosis of NSCLP. Demographic and clinical variables were collected for patients and a control group without NSCLP from the 2018 National Survey of Children’s Health (NSCH) database.Results:A total of 617 patients with NSCLP subjects and 29,147 NSCH participants were included. Among orofacial clefts, 45.2% were unilateral cleft lip and palate, followed by isolated cleft palate (30%), bilateral cleft lip and palate (16.4%), and isolated cleft lip (8.4%). NSCLP children with isolated cleft lip (odds ratio [OR]: 3.97), unilateral cleft lip and palate (OR: 2.17) and bilateral cleft lip and palate (OR: 2.91) had significantly higher odds of being diagnosed with attention-deficit hyperactivity disorder than the NSCH cohort. Rates of autism/pervasive developmental disorder were higher in children with isolated cleft lip than cleft lip and palate (11.5% versus 4.7%, P = 0.06), but this association was not significant. Children with isolated cleft palate had higher rates of intellectual disability, speech delay, global developmental delay, cerebral palsy, and hearing loss compared with the NSCH cohort (P < 0.05).Conclusions:Higher rates of attention disorders and developmental delays in children with NSCLP highlight the importance of proper risk assessment and multidisciplinary management for this population.

Behavioral assessment of auditory processing disorder in children with non-syndromic cleft lip and/or palate

Genomewide Scan for Nonsyndromic Cleft Lip and Palate in Multigenerational Indian Families Reveals Significant Evidence of Linkage at 13q33.1-34

The aim of this study was to evaluate the association of the polymorphisms in TCN2 (rs1801198) gene and in MTRR (rs1801394) gene with nonsyndromic cleft lip and/or palate (NSCL/P) in a Brazilian population. Genomic DNA was extracted from buccal cells. The polymorphisms in TCN2 (rs1801198) and MTRR (rs1801394) genes were genotyped by carrying out real-time PCR and Taqman assay. Chi-square test was used to determine the association between genotype and allele frequencies with NSCL/P and NSCL/P subgroups (cleft lip only, cleft lip and palate, and cleft palate only). Eight hundred and sixty seven unrelated individuals (401 cases with NSCL/P and 466 individuals without cleft) were evaluated. Genotype distributions of TCN2 and MTRR polymorphisms were in Hardy-Weinberg equilibrium. The TCN2 polymorphic genotype GG was identified in 16.7% of the NSCL/P group and in 14.1% of the non-cleft group (p>0.05). Similarly, the frequency of MTRR genotype (GG) was similar in NSCL/P group (15.5%) and control group (17.8%) (p>0.05). Multivariate analysis showed an association between MTRR and the subgroup that the mother smoked during pregnancy (p=0.039). Our findings did not demonstrate an association between TCN2 polymorphisms and NSCL/P, however suggests an association between MTRR and NSCL/P etiology.

Background:Nonsyndromic cleft lip and/or palate (NSCL/P) is the most common orofacial birth defect, often attributed to ethnic and environmental differences. Up to now, linkage analyses and genome-wide association studies have identified several genomic susceptibility regions for NSCL/P. The WNT genes including WNT3 are strong candidates for NSCL/P, since they are involved in regulating mid-face development and upper lip fusion. This study tested association of the WNT3 polymorphisms, rs-3809857 G/T and rs9890413 G/A, with the risk of NSCL/P in a population of Iranian infants.Methods:The allelic and genotypic frequencies for each participant were determined in 113 unrelated Iranian subjects with NSCL/P and 220 control subjects using PCR and restriction fragment length polymorphism (RFLP) methods. A p-value of ≤0.05 was considered statistically significant.Results:The WNT3 rs3809857 GT genotype was significantly lower (p=0.039, OR=0.55, 95% CI=0.30–0.97) in the NSCL/P (21.2%) than the control group (30.42%). For the WNT3 rs9890413 G/A polymorphism, neither genotype nor allele frequencies were significantly different between the case and control groups.Conclusion:Our results indicated that the WNT3 rs3809857 GT genotype may have a protective effect against NSCL/P in Iranian population.

To determine the association of single-nucleotide polymorphisms (SNPs) in genes related to craniofacial development, which were previously identified as susceptibility signals for nonsyndromic oral clefts, in Brazilians with nonsyndromic cleft lip and/or palate (NSCL/P). The SNPs rs748044 (TNP1), rs1106514 (MSX1), rs28372960, rs15251 and rs2569062 (TCOF1), rs7829058 (FGFR1), rs1793949 (COL2A1), rs11653738 (WNT3), and rs242082 (TIMP3) were assessed in a family-based transmission disequilibrium test (TDT) and a structured case-control analysis based on the individual ancestry proportions. The SNPs were initially analyzed by TDT, and polymorphisms showing a trend toward excess transmission were subsequently studied in an independent case-control sample. The study sample consisted of 189 case-parent trios of nonsyndromic cleft lip with or without cleft palate (NSCL±P), 107 case-parent trios of nonsyndromic cleft palate (NSCP), 318 isolated samples of NSCL±P, 189 isolated samples of NSCP, and 599 healthy controls. Association of alleles with NSCL/P pathogenesis. Preferential transmission of SNPs rs28372960 and rs7829058 in NSCL±P trios and rs11653738 in NSCP trios (P = .04) were observed, although the structured case-control analysis did not confirm these associations. The haplotype T-C-C formed by TCOF1 SNPs rs28372960, rs15251, and rs2569062 was more frequently transmitted from healthy parents to NSCL±P offspring, but the P value (P = .01) did not withstand Bonferroni correction for multiple tests. With the modest associations, our results do not support the hypothesis that TNP1, MSX1, TCOF1, FGFR1, COL2A1, WNT3, and TIMP3 variants are risk factors for nonsyndromic oral clefts in the Brazilian population.