Abstract
Objectives
Folate plays key role during embryogenesis. It is proved that folate deficiency is a risk factor for birth defects mainly exerting its effects through DNA methylation. Here, we want to discover the epigenetic process of DNA methylation in the regulation of miRNAs and thereby regulates neural tube closure through actions on target genes.
Methods
40 fetuses with spina bifida and 46 control fetuses from Shanxi Province in China were enrolled to study miR-324–5p alternations with folate deficiency. Meanwhile, mice model with folate deficiency diet were built to study the effects and possible mechanisms of pregnancy folate deficiency. Overexpressed miR-324–5p zebras model were used to detected the role of miR-324–5p during embryogenesis. And disturbed folic acid metabolism cell and mice model were used to confirm the mechanisms of miR-324–5p in neural tube defects induced by folate deficiency.
Results
We observed decreased miR-324–5p methylation in the brains of individuals with spina bifida. Hypomethylation of miR-324–5 increased the risk of spina bifida, with an odds ratio of 7.0 (95% CI: 2.10–23.36; P < 0.0001). A positive correlation between miR-324–5p methylation and folate levels were verified, and higher folate concentration reversed miR-324–5p alternations. We also observed increased miR-324–5p expression in the folate deficiency group, with concomitant decreased expression of its putative target genes, GLI1 and SMO. Furthermore, overexpression of miR-324–5p induced neural tube defects with inhibition of cell proliferation and migration. Similarly, we confirmed suppression of GLI1 and SMO expression in the brain of individuals with spina bifida.
Conclusions
The effect of folate deficiency on neural tube development may be mediated by miR-324–5p and its target genes.
Funding Sources
The National Natural Science Fund of China (81,670,802/H0724), National Key Research and Developmwnt Program (2016YFC1000502).