No effect of folic acid supplementation on global DNA methylation in men and women with moderately elevated homocysteine

Audrey Y Jung, Yvo Smulders, Petra Verhoef, Frans J Kok, Henk Blom, Robert M Kok, Ellen Kampman, Jane Durga, Audrey Y Jung, Yvo Smulders, Petra Verhoef, Frans J Kok, Henk Blom, Robert M Kok, Ellen Kampman, Jane Durga

Abstract

A global loss of cytosine methylation in DNA has been implicated in a wide range of diseases. There is growing evidence that modifications in DNA methylation can be brought about by altering the intake of methyl donors such as folate. We examined whether long-term daily supplementation with 0.8 mg of folic acid would increase global DNA methylation compared with placebo in individuals with elevated plasma homocysteine. We also investigated if these effects were modified by MTHFR C677T genotype. Two hundred sixteen participants out of 818 subjects who had participated in a randomized double-blind placebo-controlled trial were selected, pre-stratified on MTHFR C677T genotype and matched on age and smoking status. They were allocated to receive either folic acid (0.8 mg/d; n = 105) or placebo treatment (n = 111) for three years. Peripheral blood leukocyte DNA methylation and serum and erythrocyte folate were assessed. Global DNA methylation was measured using liquid chromatography-tandem mass spectrometry and expressed as a percentage of 5-methylcytosines versus the total number of cytosine. There was no difference in global DNA methylation between those randomized to folic acid and those in the placebo group (difference = 0.008, 95%CI = -0.05,0.07, P = 0.79). There was also no difference between treatment groups when we stratified for MTHFR C677T genotype (CC, n = 76; CT, n = 70; TT, n = 70), baseline erythrocyte folate status or baseline DNA methylation levels. In moderately hyperhomocysteinemic men and women, long-term folic acid supplementation does not increase global DNA methylation in peripheral blood leukocytes.ClinicalTrials.gov NCT00110604.

Conflict of interest statement

Competing Interests: The authors have read the journal's policy and have the following conflicts: Petra Verhoef works at Unilever R&D in Vlaardingen, The Netherlands. Jane Durga works at Nestlé Research Center in Lausanne, Switzerland. The work at both food companies entails examining the health benefits of a variety of food ingredients, including folic acid. Unilever and Nestlé market food products, some of which are enriched with folic acid. This does not alter the author's adherence to all the PLoS ONE policies on sharing data and materials. All other authors declare that no competing interests exist.

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