Abstract
The 677 C → T polymorphism in the 5-10 methylenetetrahydrofolate reductase (MTHFR) gene has been associated with nonsyndromic cleft lip with or without cleft palate (CL/P) in some populations, but not others. Previous studies (ie, case–control and transmission disequilibrium tests (TDT)) in Brazilian families with CL/P have been unable to replicate this putative association. However, our group observed a lower proportion of CT heterozygotes among the mothers of CL/P probands, suggesting that the maternal genotype for this polymorphism might influence predisposition to CL/P. In order to further examine this issue, we performed a case–control study of the 677 C → T/MTHFR polymorphism in families with CL/P ascertained in two regions of Brazil: 172 from São Paulo (SP) and 252 from Ceará (CE). The control samples included 243 individuals from SP and 401 from CE. TDT was carried out in 102 patients with CL/P and their parents. No evidence of an association was observed between the 677 C → T/MTHFR polymorphism and CL/P using the case–control design, while borderline significance was obtained with the TDT (P=0.055). We have also looked for an interaction between maternal MTHFR genotypes and the propositi offspring's genotypes at two candidate susceptibility loci for CL/P, TGFA and BCL3. Interestingly, we observed an interaction between the maternal MTHFR and offspring's BCL3 genotypes (OR: 2.3; 95% CI: 1.1–4.8; P=0.03) but not with the offspring's TGFA genotypes. Therefore, our results reinforce the idea that the maternal MTHFR genotype plays a significant role in susceptibility to CL/P, but its teratogenic effect depends on the genotype of the offspring.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Mossey PA, Little J : Epidemiology of oral clefts: an international perspective; in Wyszynski DF (ed): Cleft lip and palate: From origin to treatment. New York: Oxford University Press; 2002, pp 127–158.
Wyszynski DF, Beaty TH, Maestri NE : Genetics of nonsyndromic oral clefts revisited. Cleft Palate Craniofac J 1996; 33: 406–417.
Gorlin RJ, Cohen Jr MM, Levin LS : Syndrome of the head and neck. New York: Oxford University Press; 2001, 4th edn. pp 850–859.
Murray JC : Gene/environment causes of cleft lip and/or palate. Clin Genet 2002; 61: 248–256.
Khoury MJ, Gomez-Farial M, Mulinare J : Does maternal cigarette smoking during pregnancy cause cleft lip and palate in offspring? Am J Dis Child 1989; 143: 333–337.
Shaw GM, Lammer EJ, Wasserman CR, O'Malley CD, Tolarova MM : Risks of orofacial clefts in children born to women using multivitamins containing folic acid periconceptionally. Lancet 1995; 354: 393–396.
Czeizel AE, Toth M, Rockenbauer M : Population based case–control study of folic acid supplementation during pregnancy. Teratology 1996; 53: 345–351.
Itikala PR, Waltkins ML, Mulinare J, Moore CA, Liu Y : Maternal multivitamin use and orofacial clefts in offspring. Teratology 2001; 63: 79–86.
Czeizel AE : Prevention of oral clefts through the use of folic acid and multivitamin supplements: evidence and gaps; in Wyszynski DF (ed): Cleft lip and palate: From origin to treatment. New York: Oxford University Press; 2002, pp 443–457.
Tolarova MM, Van Rooij M, Pastor M et al: A common mutation in the MTHFR gene is a risk factor for nonsyndromic cleft lip and palate anomalies. Am J Hum Genet 1998; 63: A27.
Shaw GM, Rozen R, Finnell RH, Todoroff K, Lammer EJ : Infant C677T mutation in MTHFR, maternal periconceptional vitamin use and cleft lip. Am J Med Genet 1998; 80: 196–198.
Mills JL, Kirke PN, Molloy AM et al: Methylenetetrahydrofolate reductase thermolabile variant and oral clefts. Am J Med Genet 1999; 86: 71–74.
Gaspar DA, Pavanello RC, Zatz M et al: Role of the C677T polymorphism at the MTHFR gene on risk to nonsyndromic cleft lip with/without cleft palate: results from a case–control study in Brazil. Am J Med Genet 1999; 87: 197–199.
Martinelli M, Scapoli L, Pezzetti F et al: Linkage analysis of the three candidate regions of chromosome 1 in nonsyndromic familial orofacial cleft. Ann Hum Genet 2001; 65: 465–471.
Prescott NJ, Winter RM, Malcolm S : Maternal MTHFR genotype contributes to the risk of nonsyndromic cleft lip and palate. J Med Genet 2002; 39: 368–369.
Jugessur A, Wilcox AJ, Lie RT et al: Exploring the effects of methylenetetrahydrofolate reductase gene variants C677T and A1298C on the risk of orofacial clefts in 261 Norwegian case-parent triads. Am J Epidemol 2003; 157: 1083–1091.
van Rooij IALM, Vermeij-Keers C, Kluijtmans LAJ et al: Does the interaction between maternal folate intake and the methylenetetrahydrofolate reductase polymorphisms affect the risk of cleft lip with or without cleft palate? Am J Epidemiol 2003; 157: 583–591.
Ardinger HH, Buetow KH, Bell GI, Bardach J, Van-Demark DR, Murray JC : Association of genetic variation of the transforming growth factor-alpha gene with cleft lip and palate. Am J Hum Genet 1989; 45: 348–353.
Stein J, Mulliken JB, Stal S et al: Nonsyndromic cleft lip with or without cleft palate: evidencce of linkage to BCL3 in 17 multigenerational families. Am J Hum Genet 1995; 57: 257–272.
Gaspar DA, Matiolli S, Pavanello RC et al: Evidence that BCL3 plays a role in the etiology of nonsyndromic oral clefts in Brazilian families. Genet Epidemiol 2002; 23: 364–374.
Wyszynski DF : Locating genes for oral clefts in humans; In Wyszynski DF (ed): Cleft lip and palate: From origin to treatment. New York: Oxford University Press; 2002, pp 255–264.
Passos-Bueno MR, Gaspar DA, Kamiya T et al: Transforming growth factor alpha (TGFA) and nonsyndromic cleft lip with or without palate (CL/P) in Brazilian patients: results of a large case–control study. Cleft Palate J 2004, in press.
Azevedo ES : Subgroup studies of black admixture within a mixed population of Bahia, Brazil. Ann Hum Genet 1980; 44: 56–60.
Miller AS, Dykes DD, Polesky HF : A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988; 16: 1215.
Richards B, Skolesky J, Shuber AP et al: Multiplex PCR amplification from the CFTR gene using DNA prepared from buccal brushes/ swabs. Hum Mol Genet 1993; 2: 159–163.
Frosst P, Blom HJ, Milos R et al: A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet 1995; 10: 111–113.
Spielman RS, MacGinnis RE, Ewens WJ : Transmission test for linkage disequilibrium: the insulin gene region and insulin-dependent diabetes mellitus (IDDM). Am J Hum Genet 1993; 52: 506–516.
Guo S, Thompson E : Performing the exact test of Hardy–Weinberg proportion for multiple alleles. Biometrics 1992; 48: 361–372.
Schneider S, Roessli D, Excoffier L : Arlequin 2.00, a software for population genetics data analyses. Geneva: Department of Anthropology and Ecology, University of Geneva; 2001.
Slatkin M, Excoffier L : Testing for linkage disequilibrium in genotypic data using the EM algorithm. Heredity 1996; 76: 377–383.
Yang Q, Khoury MJ, Sun F, Flanders WD : Case-only design to measure gene–gene interaction. Epidemiology 1999; 10: 167–170.
Arruda VR, Siqueira LH, Gonçalves MS et al: Prevalence of the mutation C677T in the methylenetetrahydrofolate reductase gene among distinct ethnic groups in Brazil. Am J Med Genet 1998; 78: 332–335.
Franco RF, Araújo AG, Guerreiro JF, Elion J, Zago M : Analysis of the C677T mutation of the methylenetetrahydrofolate reductase gene in different ethnic groups. Thromb Haemost 1998; 79: 119–121.
Perez ABA, DAlmeida V, Vergani N, Oliveira AC, Lima FT, Brunoni D : Methylenetetrahydrofolate reductase (MTHFR): Incidence of mutations C677T and A1298C in Brazilian population and its correlation with plasma homocysteine levels in spina bifida. Am J Med Genet 2003; 119A: 20–25.
Rozen R, Fraser FC, Shaw G : Decreased proportion of female newborn infants homozygous for the 677C → T mutation in methylenetetrahydrofolate reductase. Am J Med Genet 1999; 83: 142–143.
Beaty TH, Hetmanski JS, Zeiger JS et al: Testing candidate genes for nonsyndromic oral clefts using a case-parent trio design. Genet Epidemiol 2002; 22: 1–11.
Shotelersuk V, Ittiwut C, Sirivan P, Angspatt A : Maternal 677CT/1298AC genotype of the MTHFR gene as a risk factor for cleft lip. J Med Genet 2003; 40: e64.
Franzoso G, Bous V, Azarenko V et al: The oncoprotein Bcl-3 can facilitate NK-kB-mediated transactivation by removing inhibiting p50 homodimers from select kB sites. EMBO J 1993; 12: 3893–3901.
Amos C, Gasser D, Hecht JT : Nonsyndromic cleft lip with or without cleft palate: new BCL3 information. Am J Hum Genet 1996; 59: 743–744.
Wyszynski DF, Maestri N, McIntosh I et al: Evidence for an association between markers on chromosome 19q and non-syndromic cleft lip with or without cleft palate in two groups of multiplex families. Hum Genet 1997; 99: 22–26.
Martinelli M, Scapoli L, Pezzetti F et al: Suggestive linkage between markers on chromosome 19q13.2 and nonsyndromic orofacial cleft malformation. Genomics 1998; 51: 177–181.
Fujita T, Nolan GP, Liou H-C, Scott ML, Baltimore D : The candidate proto-oncogene bcl-3 encodes a transcriptional coactivator that activates through NF-kB p50 homodimers. Genes Dev 1993; 7: 1354–1363.
Wong W, Eskes TKAB, Kuijpers-Jagtman AM et al: Nonsyndromic orofacial clefts: assocaition with maternal hyperhomocysteinemia. Teratology 60: 253–257.
Knott L, Hartridge T, Brown NL, Mansell JP, Sandy JR : Homocysteine oxidation and apoptosis: a potential cause of cleft palate. In Vitro Cell. Dev. Biol. Anim 2003; 39: 98–115.
Acknowledgements
We thank all of the patients and their relatives; Andréa Sertié for reading the manuscript; Carlos Maranduba for help on the analysis of the SNuPE; Constância G Urbani for secretarial help; Faculty of Dentistry at University of São Paulo, Association of Orofacial Clefts of Concórdia, Santa Catarina, and Hospital Albert Sabin, Ceará, Brazil for referring some patients. This work was supported by grants from FAPESP/CEPID, PRONEX, CNPq (except DFW). Part of this work was funded by a contract from the Massachusetts Center for Birth Defects Research and Prevention of the Massachusetts Department of Public Health (to DFW) and by an Etiology Grant of the Cleft Palate Foundation (to DFW).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gaspar, D., Matioli, S., Pavanello, R. et al. Maternal MTHFR interacts with the offspring's BCL3 genotypes, but not with TGFA, in increasing risk to nonsyndromic cleft lip with or without cleft palate. Eur J Hum Genet 12, 521–526 (2004). https://doi.org/10.1038/sj.ejhg.5201187
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.ejhg.5201187
Keywords
This article is cited by
-
Genetic markers for non-syndromic orofacial clefts in populations of European ancestry: a meta-analysis
Scientific Reports (2022)
-
Methylenetetrahydrofolate reductase C677T polymorphism is not associated with the risk of nonsyndromic cleft lip/palate: An updated meta-analysis
Scientific Reports (2020)
-
Strong Association of C677T Polymorphism of Methylenetetrahydrofolate Reductase Gene With Nosyndromic Cleft Lip/Palate (nsCL/P)
Indian Journal of Clinical Biochemistry (2018)
-
Association between MTHFR C677T polymorphism and congenital heart disease
Herz (2015)
-
Oral cleft prevention program (OCPP)
BMC Pediatrics (2012)
