368

LETTER TO JMG

J Med Genet: first published as 10.1136/jmg.39.5.368 on 1 May 2002. Downloaded from http://jmg.bmj.com/ on 7 October 2018 by guest. Protected by copyright.
Maternal MTHFR genotype contributes to the risk of
non-syndromic cleft lip and palate
N J Prescott, R M Winter, S Malcolm
.............................................................................................................................

J Med Genet 2002;39:368–369

R
ecently, we reported a whole genome scan in sib pairs
with non-syndromic cleft lip and palate (CLP), highlight- Table 1 Transmission disequilibrium test of the
ing several regions as possible susceptibility loci, one of MTHFR 677T variant allele
which is situated on 1p36.1 This region is of particular interest Transmitted Not transmitted χ2
in CLP as it harbours the gene encoding MTHFR, an enzyme
Maternal 44 52 0.67
fundamental in the metabolism of the biologically active form
Paternal 46 55 0.8
of folic acid. Dietary folic acid deficiency has been considered Total 90 107 1.46
as a candidate environmental factor in the aetiology of CLP in
several studies. A study of CL/P cases born in the Czech
Republic suggested that high dose (10 mg) folic acid
supplements taken by pregnant mothers could decrease their
chance of having a second affected CL/P child by 25-65%.2
Similar findings were reported in a case-control study in Cali- Table 2 MTHFR genotype frequencies subdivided by
fornia for a much lower dose supplementation (<1 mg).3 Also, normal and affected/carrier parents
evidence provided by a Hungarian prospective cohort study MTHFR genotype
and a study of the Hungarian Case-Control Surveillance of
Congenital Anomalies data set showed that high dose folic No CC CT TT
acid supplementation during the critical stages of craniofacial Mothers 226 103 (45%) 96 (42%) 27 (12%)
development was the most effective at reducing the occur- Affected 19 6 (31%) 7 (36%) 6 (31%)
rence of oral clefting.4 Healthy 207 97 (46%) 89 (43%) 21 (10%)
The MTHFR gene has a functional variant owing to the Fathers 210 90 (42%) 93 (44%) 27 (13%)
Affected 25 11 (44%) 9 (36%) 5 (20%)
C677T substitution. This leads to a reduced activity of the
Healthy 185 79 (43%) 84 (45%) 22 (12%)
enzyme after heating and homozygotes for this heat labile
variant have raised plasma homocysteine5 and an increased
risk for NTDs.6 Several case-control studies have attempted to
implicate this polymorphism in clefting aetiology but results
have not been encouraging. Associations have only been found
in small studies that lack corroboration when larger groups cohort showed no distortion from Hardy-Weinberg equilib-
are tested.7–10 rium, but maternal family history was considerably lower in
We have established MTHFR genotypes in 243 CLP affected our population with 19/226 (8%) mothers also being affected
subjects and their parents (226 mothers and 210 fathers) in and no others having any known family history. When fami-
order to determine if genetic susceptibility to folate deficiency lies were subdivided by parental affection status a significant
may play a role in the development of oral clefts. Families were distortion in Hardy-Weinberg equilibrium (χ2=6.07, p=0.018)
recruited from the white UK population via the Great Ormond was observed with an increased frequency of TT (31%) in the
Street Children’s Hospital, London. Appropriate ethical ap- genotypes of affected mothers of affected probands when
proval was obtained from the Great Ormond Street NHS Trust compared to healthy mothers (12%) (table 2). No such
Research Ethics Committee (No 94CG01). TDT analysis was relationship could be found with TT genotype in affected
carried out on subjects with heterozygous informative parents fathers of affected offspring. Overall this gave an odds ratio of
but showed no evidence for the association of CLP with the 4.09 (1.32-11.57) and relative risk of 3.11 (1.27-6.15) when
MTHFR T allele (table 1). Previous studies regarding the affected mothers have the TT genotype, which dropped to 1.91
MTHFR heat labile variant indicate that TT homozygotes have (0.73-5.61) and 1.29 (0.85-1.66), respectively, when hetero-
low normal serum folate and, therefore, genotype frequencies zygous mothers were also considered as carriers allowing for a
were calculated to ensure Hardy-Weinberg equilibrium among dominant model. This effect would not be observed in a TDT
affected subjects and their parents. All proband genotype fre- study as it involves homozygous parents, who are automati-
quencies obeyed Hardy-Weinberg equilibrium; C and T allele cally dropped from TDT analysis, as they are uninformative.
frequencies were 0.66 and 0.34, respectively, and comparable Consideration of paternal family history combined with
to control population frequencies in independent studies in maternal MTHFR genotype did not show any effect, indicating
the UK.11 12 that this interaction is strongest when both contributing fac-
A recent report by Martinelli et al13 observed an increase in tors (family history and MTHFR TT) are coincident through
TT homozygotes among mothers of CLP subjects indicating a the mother.
possible role for the influence of maternal genotype on fetal These findings indicate that maternal folate deficiency may
folate status with a risk ratio of 2.51 (1.00-6.14).13 The authors be a contributing factor to cleft aetiology when maternal fam-
also commented on the high degree of maternal family history ily history is observed and offer a possible explanation for the
of clefting in this cohort with approximately 50% of mothers conflicting results of previous studies investigating MTHFR
either themselves being affected or having an affected parent. genotype in relation to CLP. Such results would also suggest
Observation of maternal genotype frequencies within our the importance of collecting parental and grandparental data

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Letters 369

Correspondence to: Dr N J Prescott, Clinical and Molecular Genetics

• We genotyped 243 parent-case triads for the 5,10- Unit, Institute of Child Health, 30 Guilford Street, London WC1N 1EH,
methylenetetrahydrofolate reductase (MTHFR) C677T UK; n.prescott@ich.ucl.ac.uk

J Med Genet: first published as 10.1136/jmg.39.5.368 on 1 May 2002. Downloaded from http://jmg.bmj.com/ on 7 October 2018 by guest. Protected by copyright.
polymorphism to determine whether this functional
variant is responsible for non-syndromic cleft lip and pal-
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Weinberg equilibrium detected an over-representation of
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ACKNOWLEDGEMENTS C677T (thermolabile alanine/valine) polymorphism in
The authors would like to thank Sharon Thomas for her help with methylenetetrahydrofolate reductase (MTHFR): its frequency and impact
sample collection. We acknowledge the financial support of the UK on plasma homocysteine concentration in different European populations.
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Authors’ affiliations 13 Martinelli M, Scapoli L, Pezzetti F, Carinci F, Carinci P, Stabellini G,
N J Prescott, R M Winter, S Malcolm, Clinical and Molecular Genetics Bisceglia L, Gombos F, Tognon M. C677T variant form at the MTHFR
Unit, Institute of Child Health, 30 Guilford Street, London WC1N 1EH, gene and CL/P: a risk factor for mothers? Am J Med Genet
UK 2001;98:357-60.

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