Nonsyndromic cleft lip with or without cleft palate (NSCLP) is a common birth defect with substantial clinical and social impact. Folate deficiency is one of the factors that have been associated with increased risk for NSCLP. Polymorphisms in folate and homocysteine pathway genes may act as susceptibility factors.
Objective
The objective of this study was to evaluate prevalence estimates of cystathionine beta-synthase (CBS) insertion of 68-bp (c.844ins68) polymorphisms and their correlation with NSCLP.
Material and Methods
A total of 236 unrelated individuals from seven Indian populations and an additional 355 cases with NSCLP and 357 controls without NSCLP were included in this study. We investigated the CBS c.844ins68 polymorphism in all samples. Genotyping was performed with polymerase chain reaction and electrophoresis. The data were statistically analyzed using the chi-square test.
Results
The CBS c.844ins68 allele is present in six of the seven populations analyzed, and allele frequencies range from 1.5% in Balija to 9.1% in Sugali populations. The CBS c.844ins68 polymorphism showed a significant protective effect on NSCLP at both genotype (WW versus WI: odds ratio [OR] = 0.54, 95% confidence interval [CI] = 0.31 to 0.95, P = .149) and allele levels (W versus I: OR = 0.56, 95% CI = 0.32 to 0.96, P = .033).
Conclusions
The current study observed significant differences in the frequency of the CBS 844ins68 allele across populations. There is a significant association between CBS c.844ins68 polymorphism and cleft lip and palate in the Indian population. Additional studies are warranted to identify the functional variants in the genes controlling homocysteine as etiological contributors to the formation of oral clefts.
AlessioA.C., SiqueiraL.H., BydlowskiS.P., HoehrN.F., Annichino-BizzacchiJ.M.Polymorphisms in the CBS gene and homocysteine, folate and vitamin B12 levels: association with polymorphisms in the MTHFR and MTRR genes in Brazilian children. Am J Med Genet A.2008; 146A: 2598–2602.
2.
AyalaC., GarciaR., CruzE., PrietoK., BermudezM.Homocysteine levels and polymorphisms of MTHFR and CBS genes in Colombian patients with superficial and deep venous thrombosis [in Spanish]. Biomedica.2010; 30: 259–267.
3.
BhaskarL.V., MurthyJ., Venkatesh BabuG.Polymorphisms in genes involved in folate metabolism and orofacial clefts. Arch Oral Biol.2011; 56: 723–737.
4.
BirnbaumS., ReutterH., MendeM., Diaz-LacavaA., HenschkeH., BergeS.J., BraumannB., LausterC., HemprichA., WenghoeferM.. A family-based association study in Central Europeans: no evidence for the cystathionine beta-synthase c.844ins68 gene variant as a risk factor for non-syndromic cleft lip and palate. Am J Med Genet A.2007; 143: 205–207.
5.
BlantonS.H., HenryR.R., YuanQ., MullikenJ.B., StalS., FinnellR.H., HechtJ.T.Folate pathway and nonsyndromic cleft lip and palate. Birth Defects Res A Clin Mol Teratol.2011; 91: 50–60.
6.
BoylesA.L., WilcoxA.J., TaylorJ.A., MeyerK., FredriksenA., UelandP.M., DrevonC.A., VollsetS.E., LieR.T.Folate and one-carbon metabolism gene polymorphisms and their associations with oral facial clefts. Am J Med Genet A.2008; 146A: 440–449.
7.
BoylesA.L., WilcoxA.J., TaylorJ.A., ShiM., WeinbergC.R., MeyerK., FredriksenA., UelandP.M., JohansenA.M., DrevonC.A.. Oral facial clefts and gene polymorphisms in metabolism of folate/one-carbon and vitamin A: a pathway-wide association study. Genet Epidemiol.2009; 33: 247–255.
8.
ChiquetB.T., HenryR., BurtA., MullikenJ.B., StalS., BlantonS.H., HechtJ.T.Nonsyndromic cleft lip and palate: CRISPLD genes and the folate gene pathway connection. Birth Defects Res A Clin Mol Teratol.2011; 91: 44–49.
9.
CubellsJ.F., KobayashiK., NagatsuT., KiddK.K., KiddJ.R., CalafellF., KranzlerH.R., IchinoseH., GelernterJ.Population genetics of a functional variant of the dopamine beta-hydroxylase gene (DBH). Am J Med Genet.1997; 74: 374–379.
10.
DasarathyJ., GrucaL.L., BennettC., ParimiP.S., DuenasC., MarczewskiS., FierroJ.L., KalhanS.C.Methionine metabolism in human pregnancy. Am J Clin Nutr.2010; 91: 357–365.
11.
DuttaS., ChatterjeeA., SinhaS., ChattopadhyayA., MukhopadhyayK.Correlation between cystathionine beta synthase gene polymorphisms, plasma homocysteine and idiopathic mental retardation in Indian individuals from Kolkata. Neurosci Lett.2009; 453: 214–218.
12.
DuttaS., SinhaS., ChattopadhyayA., GangopadhyayP.K., MukhopadhyayJ., SinghM., MukhopadhyayK.Cystathionine beta-synthase T833C/844INS68 polymorphism: a family-based study on mentally retarded children. Behav Brain Funct.2005; 1: 25.
13.
EussenS.J., VollsetS.E., HustadS., MidttunO., MeyerK., FredriksenA., UelandP.M., JenabM., SlimaniN., FerrariP.. Vitamins B2 and B6 and genetic polymorphisms related to one-carbon metabolism as risk factors for gastric adenocarcinoma in the European prospective investigation into cancer and nutrition. Cancer Epidemiol Biomarkers Prev.2010; 19: 28–38.
14.
FrancoR., MaffeiF., LourencoD., PiccinatoC., MorelliV., ThomaziniI., ZagoM.The frequency of 844ins68 mutation in the cystathionine beta-synthase gene is not increased in patients with venous thrombosis. Haematologica.1998a; 83: 1006–1008.
15.
FrancoR.F., ElionJ., LavinhaJ., KrishnamoorthyR., TavellaM.H., ZagoM.A.Heterogeneous ethnic distribution of the 844ins68 in the cystathionine beta-synthase gene. Hum Hered.1998b; 48: 338–342.
16.
GalbiattiA.L., RuizM.T., RaposoL.S., ManigliaJ.V., Pavarino-BertelliE.C., Goloni-BertolloE.M.The association between CBS 844ins68 polymorphism and head and neck squamous cell carcinoma risk: a case-control analysis. Arch Med Sci.2010; 6: 772–779.
17.
GeY., JensenT., JamesS.J., BectonD.L., MasseyG.V., WeinsteinH.J., RavindranathY., MatherlyL.H., TaubJ.W.High frequency of the 844ins68 cystathionine-beta-synthase gene variant in Down syndrome children with acute myeloid leukemia. Leukemia.2002; 16: 2339–2341.
18.
GolimbetV., KorovaitsevaG., AbramovaL., KaledaV.The 844ins68 polymorphism of the cystathionine beta-synthase gene is associated with schizophrenia. Psychiatry Res.2009; 170: 168–171.
19.
GrobelnyB.T., DucruetA.F., DeRosaP.A., KotchetkovI.S., ZachariaB.E., HickmanZ.L., FernandezL., NarulaR., ClaassenJ., LeeK.. Gain-of-function polymorphisms of cystathionine beta-synthase and delayed cerebral ischemia following aneurysmal subarachnoid hemorrhage. J Neurosurg.2011; 115: 101–107.
20.
GrossmannR., SchwenderS., GeisenU., SchambeckC., MeratiG., WalterU.CBS 844ins68, MTHFR TT677 and EPCR 4031ins23 genotypes in patients with deep-vein thrombosis. Thromb Res.2002; 107: 13–15.
21.
HozyaszK.K., MostowskaA., Szaflarska-PoplawskaA., LianeriM., JagodzinskiP.P.Polymorphic variants of genes involved in homocysteine metabolism in celiac disease. Mol Biol Rep.2012; 39: 3123–3130.
22.
KoutmosM., KabilO., SmithJ.L., BanerjeeR.Structural basis for substrate activation and regulation by cystathionine beta-synthase (CBS) domains in cystathionine {beta}-synthase. Proc Natl Acad Sci U S A.2010; 107: 20958–20963.
23.
LiA.F., ZhengH., XuY.M., ZhaoX.J., ZhangX.M.The association between gene polymorphisms of homocysteine metabolism-related enzymes and ischemic cerebrovascular diseases in Chinese Henan Han population. Life Sci J.2012; 9: 403–408.
24.
MartinelliM., MasieroE., CarinciF., MorselliP.G., PezzettiF., ScapoliL.New evidence for the role of cystathionine beta-synthase in nonsyndromic cleft lip with or without cleft palate. Eur J Oral Sci.2011; 119: 193–197.
25.
MostowskaA., HozyaszK.K., WojcickiP., DziegelewskaM., JagodzinskiP.P.Associations of folate and choline metabolism gene polymorphisms with orofacial clefts. J Med Genet.2010; 47: 809–815.
26.
MunkeM., KrausJ.P., OhuraT., FranckeU.The gene for cystathionine beta-synthase (CBS) maps to the subtelomeric region on human chromosome 21q and to proximal mouse chromosome 17. Am J Hum Genet.1988; 42: 550–559.
27.
MurthyJ., BhaskarL.Current concepts in genetics of nonsyndromic clefts. Indian J Plast Surg.2009; 42: 68–81.
28.
OttN., GeddertH., SarbiaM.Polymorphisms in methionine synthase (A2756G) and cystathionine beta-synthase (844ins68) and susceptibility to carcinomas of the upper gastrointestinal tract. J Cancer Res Clin Oncol.2008; 134: 405–410.
29.
PepeG., VanegasO.C., RickardsO., GiustiB., ComeglioP., BrunelliT., MarcucciR., PriscoD., GensiniG.F., AbbateR.World distribution of the T833C/844INS68 CBS in cis double mutation: a reliable anthropological marker. Hum Genet.1999; 104: 126–129.
30.
PoppR., CrisanT., RotarI., FarcasM., TrifaA., MilitaruM., PopI.Cystathionine β-synthase 844ins68 genetic polymorphism in spontaneous abortion susceptibility. Appl Med Inform.2011; 29: 34–40.
31.
PrudovaA., BaumanZ., BraunA., VitvitskyV., LuS.C., BanerjeeR.S-adenosylmethionine stabilizes cystathionine beta-synthase and modulates redox capacity. Proc Natl Acad Sci U S A.2006; 103: 6489–6494.
32.
RatnamS., MacleanK.N., JacobsR.L., BrosnanM.E., KrausJ.P., BrosnanJ.T.Hormonal regulation of cystathionine beta-synthase expression in liver. J Biol Chem.2002; 277: 42912–42918.
33.
ReltonC.L., WildingC.S., PearceM.S., LafflingA.J., JonasP.A., LynchS.A., TawnE.J., BurnJ.Gene-gene interaction in folate-related genes and risk of neural tube defects in a UK population. J Med Genet.2004; 41: 256–260.
34.
RubiniM., BrusatiR., GarattiniG., MagnaniC., LivieroF., BianchiF., TarantinoE., MasseiA., PollastriS., CarturanS.. Cystathionine beta-synthase c.844ins68 gene variant and non-syndromic cleft lip and palate. Am J Med Genet A.2005; 136A: 368–372.
35.
SambrookJ., RussellD.W.Molecular Cloning: A Laboratory Manual.3rd ed.Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;2001.
36.
SenemarS., DoroudchiM., PezeshkiA.M., BazrgarM., Torab-JahromiA., GhaderiA.Frequency of cystathionine beta-synthase 844INS68 polymorphism in Southern Iran. Mol Biol Rep.2009; 36: 353–356.
37.
ShannonB., GnanasampanthanS., BeilbyJ., IacopettaB.A polymorphism in the methylenetetrahydrofolate reductase gene predisposes to colorectal cancers with microsatellite instability. Gut.2002; 50: 520–524.
38.
SongK.S., ChoiJ.R., KimH.K., GeeS.H., ShimW.H.Presence of 844ins68 in the cystathionine beta-synthase gene in Asians (Koreans). Thromb Haemost.2001; 86: 1130.
39.
SpeerM.C., NyeJ., McLoneD., WorleyG., MelvinE.C., VilesK.D., FranklinA., DrakeC., MackeyJ., GeorgeT.M.Possible interaction of genotypes at cystathionine beta-synthase and methylenetetrahydrofolate reductase (MTHFR) in neural tube defects. NTD Collaborative Group. Clin Genet.1999; 56: 142–144.
40.
SperandeoM.P., de FranchisR., AndriaG., SebastioG.A 68-bp insertion found in a homocystinuric patient is a common variant and is skipped by alternative splicing of the cystathionine beta-synthase mRNA. Am J Hum Genet.1996; 59: 1391–1393.
41.
SummersC.M., HammonsA.L., MitchellL.E., WoodsideJ.V., YarnellJ.W., YoungI.S., EvansA., WhiteheadA.S.Influence of the cystathionine beta-synthase 844ins68 and methylenetetrahydrofolate reductase 677C>T polymorphisms on folate and homocysteine concentrations. Eur J Hum Genet.2008; 16: 1010–1013.
42.
TsaiM.Y., BignellM., SchwichtenbergK., HansonN.Q.High prevalence of a mutation in the cystathionine beta-synthase gene. Am J Hum Genet.1996; 59: 1262–1267.
43.
TsaiM.Y., GargU., KeyN.S., HansonN.Q., SuhA., SchwichtenbergK.Molecular and biochemical approaches in the identification of heterozygotes for homocystinuria. Atherosclerosis.1996; 122: 69–77.
44.
UrreiztiR., AsteggianoC., VilasecaM.A., CorbellaE., PintoX., GrinbergD., BalcellsS.A CBS haplotype and a polymorphism at the MSR gene are associated with cardiovascular disease in a Spanish case-control study. Clin Biochem.2007; 40: 864–868.
45.
WeinerA.S., BoyarskikhU.A., VoroninaE.N., SeleznevaI.A., SinkinaT.V., LazarevA.F., PetrovaV.D., FilipenkoM.L.Polymorphisms in the folate-metabolizing genes MTR, MTRR, and CBS and breast cancer risk. Cancer Epidemiol.2012; 36: e95–e100.
46.
YakubM., MotiN., ParveenS., ChaudhryB., AzamI., IqbalM.P.Polymorphisms in MTHFR, MS and CBS genes and homocysteine levels in a Pakistani population. PLoS ONE.2012; 7: e33222.
47.
ZhangG., DaiC.Gene polymorphisms of homocysteine metabolism-related enzymes in Chinese patients with occlusive coronary artery or cerebral vascular diseases. Thromb Res.2001; 104: 187–195.
