Folate and Clefts of the Lip and Palate—A U.K.-Based Case-Control Study: Part II: Biochemical and Genetic Analysis

Abstract

Objective:

To investigate associations between nonsyndromic oral clefts and biochemical measures of folate status and the MTHFR C677T variant in the United Kingdom, where there has been no folic acid fortification program.

Method:

Dietary details were obtained from the mothers of 112 cases of cleft lip with or without cleft palate (CL±P), 78 cleft palate only (CP) cases, and 248 unaffected infants. Infant and parental MTHFR C677T genotype was determined. Red blood cell (RBC) and serum folate and homocysteine levels were assessed in 12-month postpartum blood samples from a subset of mothers. The data were analyzed by logistic and log-linear regression methods.

Results:

There was an inverse association between CL±P and maternal MTHFR CT (odds ratio [OR] = 0.5, 95% confidence interval [CI] = 0.31–0.95) and TT (OR = 0.6, 95% CI = 0.21–1.50) genotypes, with similar risk estimates for CP. There was no clear association with infant MTHFR genotype. Higher levels of maternal postpartum RBC and serum folate were associated with a lower risk for CL±P and an increased risk for CP. Higher levels of serum homocysteine were associated with a slightly increased risk for both CL±P and CP.

Conclusion:

While the inverse relation between the mother's having the MTHFR C677T variant and both CL±P and CP suggests perturbation of maternal folate metabolism is of etiological importance, contrasting relations between maternal postpartum levels of RBC and serum folate by type of cleft are difficult to explain.

Keywords

case control folic acid MTHFR cleft lip cleft palate polymorphism red cell folate serum folate homocysteine

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References

Badovinac

R.L.

Werler

M.M.

Williams

P.L.

Kelsey

K.T.

Hayes

Folic acid–containing supplement consumption during pregnancy and risk for oral clefts: a meta-analysis.

Birth Defects Res A. 2007; 79: 8–15.

Blanton

S.H.

Kolle

B.S.

Hecht

J.T.

Mulliken

J.B.

Martin

E.R.

No evidence supporting MTHFR as a risk factor in the development of familial NSCLP.

Am J Med Genet. 2000; 92: 370–371.

Blanton

S.H.

Patel

Hecht

J.T.

Mulliken

J.B.

MTHFR is not a risk factor in the development of isolated nonsyndromic cleft lip and palate.

Am J Genet. 2002; 110: 404–405.

Botto

L.D.

Yang

5,10-Methylenetetrahydrofolate reductase gene variants and congenital anomalies: a HuGE review.

Am J Epidemiol. 2000; 151: 862–877.

Chevrier

Perret

Bahuau

Zhu

Nelva

Herman

Francannet

Robert-Gnansia

Finnell

R.H.

Cordier

Fetal and maternal MTHFR C677T genotype, maternal folate intake and the risk of nonsyndromic oral clefts.

Am J Med Genet A. 2007; l143A: 248–257.

Devine

C.M.

Bove

C.F.

Olson

C.M.

Continuity and change in women's weight orientations and lifestyle practices through pregnancy and the postpartum period: the influence of life course trajectories and transitional events.

Soc Sci Med. 2000; 50: 567–582.

Gaspar

D.A.

Matioli

S.R.

de Cassia Pavanello

Araujo

B.C.

Alonso

Wyszynski

Passos-Bueno

M.R.

Maternal MTHFR interacts with the offspring's BCL3 genotypes, but not with TGFα, in increasing risk to nonsyndromic cleft lip with or without cleft palate.

Eur J Hum Genet. 2004; 12: 521–526.

Gaspar

D.A.

Pavanello

R.C.

Zatz

Passos-Bueno

M.R.

Andre

Steman

Wyszynski

D.F.

Matiolli

S.R.

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.

Girelli

Friso

Trabetti

Olivieri

Russo

Pessotto

Faccini

Pignatti

P.F.

Mazzucco

Corrocher

Methylenetetrahydrofolate reductase C677T mutation, plasma homocysteine, and folate in subjects from northern Italy with or without angiographically documented severe coronary atherosclerotic disease: evidence for an important genetic-environmental interaction.

Blood. 1998; 91: 4158–4163.

10.

Hashmi

S.S.

Waller

D.K.

Langlois

Canfield

Hecht

J.T.

Prevalence of nonsyndromic oral clefts in Texas: 1995–1999.

Am J Med Genet A. 2005; 134: 368–372.

11.

Hill

A.E.

FitzPatrick

D.R.

MS-PCR assay to detect 677C→T mutation in the 5,10-methylenetetrahydrofolate reductase gene.

J Inherit Metab Dis. 1998; 21: 694–695.

12.

Holland

Welch

A.A.

Unwin

I.D.

McCance and Widdowson's The Composition of Foods. 5th revised and extended edition. Cambridge, UK: Royal Society of Chemistry and Ministry of Agriculture, Fisheries and Food; 1991.

13.

Hosmer

D.W.

Lemeshow

Applied Logistic Regression. New York: Wiley; 1989.

14.

Hsieh

C.C.

Maisonneuve

Boyle

Macfarlane

G.J.

Robertson

Analysis of quantitative data by quantiles in epidemiologic studies: classification according to cases, noncases, or all subjects?

Epidemiology. 1991; 2: 137–140.

15.

Jacques

P.F.

Bostom

A.G.

Williams

R.R.

Ellison

R.C.

Eckfeldt

J.H.

Rosenberg

I.H.

Selhub

Rozen

Relation between folate status, a common mutation in methylenetetrahydrofolate reductase, and plasma homocysteine concentrations.

Circulation. 1996; 93: 7–9.

16.

Jugessur

Wilcox

A.J.

Lie

R.T.

Murray

J.C.

Taylor

J.A.

Ulvik

Drecon

C.A.

Vindenes

H.A.

Abyholm

F.E.

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 Epidemiol. 2003; 157: 1083–1091.

17.

Khoury

M.J.

Stewart

Beaty

T.H.

The effect of genetic susceptibility on causal inference in epidemiologic studies.

Am J Epidemiol. 1987; 126: 561–567.

18.

Lareu

M.V.

Ruiz-Ponte

Genotyping SNPs with the LightCycler.

Methods Mol Biol. 2005; 297: 127–140.

19.

Leck

Iles

C.A.

Sharman

I.M.

Toe

Wadsworth

G.R.

Maternal diet and nutrition during early pregnancy and after delivery in North London. In: Dobbing

, ed. Prevention of Spina Bifida and Other Neural Tube Defects. London: Academic Press; 1983: 197–215.

20.

Little

Cardy

Munger

R.G.

Tobacco smoking and oral clefts: a meta-analysis.

Bull World Health Organ. 2004; 82: 213–218.

21.

Little

Gilmour

Mossey

P.A.

FitzPatrick

Cardy

Clayton-Smith

Fryer

A.E.

on behalf of the ITS MAGIC collaboration. Folate and clefts of the lip and palate—a UK based case-control study: part I: dietary and supplemental folate. Cleft Palate Craniofac J. 2008; 45.

22.

Stampfer

M.J.

Christensen

Giovannucci

Hunter

D.J.

Chen

Willett

W.C.

Selhub

Hennekens

C.H.

Gravel

. A polymorphism of the methionine synthase gene: association with plasma folate, vitamin B12, homocyst(e)ine, and colorectal cancer risk. Cancer Epidemiol Biomarkers Prev. 1999; 8: 825–829.

23.

Stampfer

M.J.

Giovannucci

Christensen

Willett

W.C.

Rozen

Methylenetetrahydrofolate reductase polymorphism, dietary interactions and risk of colorectal cancer.

Cancer Res. 1997; 57: 1098–1102.

24.

Mannino

D.M.

Mulinare

Ford

E.S.

Schwartz

Tobacco smoke exposure and decreased serum and red blood cell folate levels: data from the Third National Health and Nutrition Examination survey.

Nicotine Tob Res. 2003; 5: 357–362.

25.

Martinelli

Scapoli

Pezzetti

Carinci

Stabellini

Bisceglia

Gombos

Tognon

C677T variant form at the MTHFR gene and CL/P: a risk factor for mothers?

Am J Med Genet. 2001; 98: 357–360.

26.

Masson

L.F.

McNeill

Tomany

T.O.

Simpson

J.A.

Peace

H.S.

Wei

Grubb

D.A.

Bolton-Smith

Statistical approaches for assessing the relative validity of a food-frequency questionnaire: use of correlation coefficients and the kappa statistic.

Public Health Nutr. 2003; 6: 313–321.

27.

Mitchell

A.A.

Cutler

D.J.

Chakravarti

Undetected genotyping errors cause apparent overtransmission of common alleles in the transmission/disequilibrium test.

Am J Hum Genet. 2003; 72: 598–610.

28.

Molloy

A.M.

Daly

Mills

J.L.

Whitehead

A.S.

Kirke

P.N.

Scott

J.M.

Thermolabile variant of 5,10-methylenetetrahydrofolate reductase associated with low red-cell folates: implications for folate intake recommendations.

Lancet. 1997; 349: 1591–1593.

29.

Molloy

A.M.

Mills

J.L.

Kirke

P.N.

Whitehead

A.S.

Weir

D.G.

Scott

J.M.

Whole-blood folate values in subjects with different methylenetetrahydrofolate reductase genotypes: differences between the radioassay and microbiological assays.

Clin Chem. 1998; 44: 186–188.

30.

Molloy

A.M.

Scott

J.M.

Microbiological assay for serum, plasma, and red cell folate using cryopreserved, microtiter plate method.

Methods Enzymol. 1997; 281: 43–53.

31.

Mostowska

Hozyasz

Jagodzinski

P.P.

Maternal MTR genotype contributes to the risk of non-syndromic cleft lip and palate in the Polish population.

Clin Genet. 2006; 69: 512–517.

32.

Munger

Maternal nutrition and oral clefts. In: Wyszynski

D.F.

, ed. Cleft Lip and Palate: From Origin to Treatment. New York: Oxford University Press; 2002: 170–192.

33.

Munger

R.G.

Sauberlich

H.E.

Corcoran

Nepomuceno

Daack-Hirsch

Solon

F.S.

Maternal vitamin B-6 and folate status and risk of oral cleft birth defects in the Philippines.

Birth Defects Res. 2004; 70: 464–471.

34.

Munger

Sauberlich

Murray

Layda

Villaneuva

Magee

Abnormal folate and retinol levels in Filipino mothers of children with orofacial clefts.

FASEB J. 1996; 10: 21–25.

35.

Narayanan

McConnell

Little

Sharp

Piyathilake

C.J.

Powers

Basten

Duthie

S.J.

Associations between two common variants C677T and A1298C in the methylenetetrahydrofolate reductase gene and measures of folate metabolism and DNA stability (strand breaks, misincorporated uracil and DNA methylation status) in human lymphocytes in vivo.

Cancer Epidemiol Biomarkers Prev. 2004; 13: 1436–1443.

36.

Nurk

Tell

G.S.

Refsum

Ueland

P.M.

Vollset

S.E.

Associations between maternal methylenetetrahydrofolate reductase polymorphisms and adverse outcomes of pregnancy: the Hordaland Homocysteine Study.

Am J Med. 2004; 117: 26–31.

37.

O'Connor

Molloy

A.M.

Daly

Scott

J.M.

Deriving a useful packed cell volume estimate from haemoglobin analysis.

J Clin Pathol. 1994; 47: 78–79.

38.

Pezzetti

Martinelli

Scapoli

Carinci

Palmieri

Marchesini

Carinci

Caramelli

Rullo

Gombos

. Maternal MTHFR variant forms increase the risk in offspring of isolated nonsyndromic cleft lip with or without cleft palate. Hum Mutat. 2004; 24: 104–105.

39.

Ray

J.G.

Meier

Vermeulen

M.J.

Wyatt

P.R.

Cole

D.E.

Association between folic acid food fortification and congenital orofacial clefts.

J Pediatr. 2003; 143: 805–807.

40.

Risch

Teng

The relative power of family-based and case-control designs for linkage disequilibrium studies of complex human diseases 1. DNA pooling.

Genome Res. 1998; 8: 1273–1288.

41.

Rozen

Genetic predisposition to hyperhomocysteinemia: deficiency of methylenetetrahydrofolate reductase (MTHFR).

Thromb Haemost. 1997; 78: 523–526.

42.

Salanti

Amountza

Ntzani

E.E.

Ioannidis

J.P.

Hardy-Weinberg equilibrium in genetic association studies: an empirical evaluation of reporting, deviations, and power.

Eur J Hum Genet. 2005; 13: 840–848.

43.

Shaw

G.M.

Rozen

Finnell

R.H.

Todoroff

Lammer

E.J.

Infant C677T mutation in MTHFR, maternal periconceptional vitamin use, and cleft lip.

Am J Med Genet. 1998; 80: 196–198.

44.

Shaw

G.M.

Todoroff

Finnell

R.H.

Rozen

Lammer

E.J.

Maternal vitamin use, infant C677T mutation in MTHFR, and isolated cleft palate risk.

Am J Med Genet. 1999; 85: 84–85.

45.

Shi

Caprau

Dagle

Christiansen

Christensen

Murray

J.C.

Application of kinetic polymerase chain reaction and molecular beacon assays to pooled analyses and high-throughput genotyping for candidate genes.

Birth Defects Res A Clin Mol Teratol. 2004; 70: 65–74.

46.

Shotelersuk

Ittiwut

Siriwan

Angspatt

Maternal 677CT/1298AC genotype of the MTHFR gene as a risk factor for cleft lip.

J Med Genet. 2003; 40: e64.

47.

van Rooij

I.A.

Vermeij-Keers

Kluijtmans

L.A.

Ocke

M.C.

Zielhuis

G.A.

Goorhuis-Brouwer

S.M.

van der Biezen

J.J.

Kuijpers-Jagtman

A.M.

Steegers-Theunissen

R.P.

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.

48.

Vieira

A.R.

Murray

J.C.

Trembath

Orioli

I.M.

Castilla

E.E.

Cooper

M.E.

Marazita

M.L.

Lennon-Graham

Speer

Studies of reduced folate carrier 1 (RFC1) A80G and 5, 10-methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms with neural tube and orofacial cleft defects.

Am J Med Genet. 2005; 135: 220–223.

49.

Wan

W.D.

Wang

L.J.

Zhou

X.P.

Zhou

D.L.

Zhang

Q.G.

Huang

J.L.

Wang

X.N.

Relationship between nonsyndromic cleft lip with or without cleft palate (NSCL/P) and genetic polymorphisms of MTHFR C677T and A1298C.

Zhonghua Zheng Xing Wai Ke Za Zhi. 2006; 22: 8–11.

50.

Weinberg

C.R.

Allowing for missing parents in genetic studies of case-parent triads.

Am J Hum Genet. 1999; 64: 1186–1193.

51.

Weinberg

C.R.

Wilcox

A.J.

Lie

R.T.

A log-linear approach to case-parent-triad data: assessing effects of disease genes that act either directly or through maternal effects and that may be subject to parental imprinting.

Am J Human Genet. 1998; 62: 969–978.

52.

Wilcox

A.J.

Lie

R.T.

Solvoll

Taylor

McConnaughey

D.R.

Vindenes

Vollset

S.E.

Drevon

C.A.

Folic acid supplements and risk of facial clefts: national population based case-control study.

BMJ. 2007; 334: 433–434.

53.

Wilcox

A.J.

Weinberg

C.R.

Lie

R.T.

Distinguishing the effects of maternal and offspring genes through studies of “case-parent triads.”

Am J Epidemiol. 1998; 148: 893–901.

54.

Willett

W.C.

Howe

G.R.

Kushi

L.H.

Adjustment for total energy intake in epidemiologic studies.

Am J Clin Nutr. 1997; 65(4 suppl): 1220S–1228S.

55.

Witte

J.S.

Gauderman

W.J.

Thomas

D.C.

Asymptotic bias and efficiency in case-control studies of candidate genes and gene-environment interactions: basic family designs.

Am J Epidemiol. 1999; 149: 693–705.

56.

Wong

W.Y.

Eskes

T.K.A.B.

Kuijpers-Jagtman

A.M.

Spauwen

P.H.M.

Steegers

E.A.P.

Thomas

C.M.

Hamel

B.C.

Blom

H.J.

Steegers-Theunissen

R.P.

Nonsyndromic orofacial clefts: association with maternal hyperhomocysteinemia.

Teratology. 1999; 60: 253–257.

57.

Yazdy

M.M.

Honein

M.A.

Xing

Reduction in orofacial clefts following folic acid fortification of the U.S. grain supply.

Birth Defects Res A Clin Mol Teratol. 2007; 79: 16–23.

58.

Zhu

Ren

Hao

Pei

Liu

Zhu

Finnell

R.H.

Variable contribution of the MTHFR C677T polymorphism to non-syndromic cleft lip and palate risk in China.

Am J Med Genet A. 2006; 140: 551–557.