Murine Methyl Donor Deficiency Impairs Early Growth in Association with Dysmorphic Small Intestinal Crypts and Reduced Gut Microbial Community Diversity

Abstract

BackgroundFolate and choline are essential methyl donor nutrients throughout the life span; however, the adverse effects of combined deficiency on early growth, intestinal epithelial morphology, and the gut microbiome remain only partially understood.ObjectivesWe investigated the effects of dietary folate and choline deficiency on early growth, small intestinal (SI) epithelial architecture, and the gut microbiota of mice. To explore potential mechanisms for adverse effects on gut epithelial morphology, we also evaluated gene expression and DNA methylation in mouse intestinal epithelial organoids (enteroids) maintained in methyl donor–deficient (MDD) conditions.MethodsPregnant dams were administered 1 of 4 diets: 1) control diet (CD−), 2) an isocaloric MDD− diet, or 3) CD+ and 4) MDD+ formulations containing 1% succinylsulfathiazole to inhibit folate-producing gut bacteria. We weaned pups to their dams’ diet at 3 wk of age and monitored body weight and tail length pre- and postweaning. We measured serum folate, SI crypt morphology, and microbiota composition at 7 wk of age.ResultsBoth MDD+ and MDD− diets impaired early ponderal and linear growth, lowered serum folate concentrations, and produced patchy areas of increased crypt depth throughout the SI. Succinylsulfathiazole increased crypt depth independently of diet. MDD or succinylsulfathiazole, alone or in combination, altered the gut microbiome, with decreased Bacteroidales and Clostridiales, increased Lactobacillales and Erysipelotrichaceae taxa, and decreased α-diversity indexes. Enteroids maintained in MDD media displayed dysmorphic crypt domains, altered expression of stem cell and secretory differentiation genes, and decreased DNA methylation of the glycosylation genes Beta-1,4-N-Acetyl-Galactosaminyltransferase-1 (B4galnt1) and Phosphoethanolamine/Phosphocholine-Phosphatase (Phospho1).ConclusionMDD impairs ponderal and linear growth in mice in association with dysmorphic SI crypts and reduced gut microbial diversity. In vitro methyl donor deficiency similarly induced dysmorphic crypts in mouse enteroids in conjunction with altered gene expression and DNA methylation.

Mucosal Genomics Implicate Lymphocyte Activation and Lipid Metabolism in Refractory Environmental Enteric Dysfunction

“…Supplementation with folate—an essential methyl donor nutrient—is an effective adjunct therapy for persistent diarrhea in children with malnutrition. 31 Further, intestinal stem cell–specific deletion of DNA methyltransferase 1 32 or a diet deficient in folate and choline 33 recapitulates several features of EED in mice. Further, the abundance of differentially methylated genes detected in our study suggests fecal intestinal epithelial cell methylation screens might be developed for EED to provide a noninvasive stool-based approached for detection and monitoring of EED, as is currently done for colorectal cancer.…”

DNA methylation as a regulator of intestinal gene expression

“…Morphological changes due to MD supplementation/deficiency have also been observed in the intestines. Silva et al, 2019 found sporadic regions throughout the small intestine with increased crypt depth in the offspring of dams submitted to a methyl deficient diet during gestation until weaning (110) .…”

How To Build a Bone: PHOSPHO1, Biomineralization, and Beyond

“…PHOSPHO1 may therefore be an appealing candidate to act within a secondary mechanism of choline homeostasis in these and other tissues. Indeed, gut enteroids maintained in choline‐deficient media exhibited hypomethylation at 3′ CpG islands within the Phospho1 gene, potentially regulating gene expression …”

“…Indeed, gut enteroids maintained in choline-deficient media exhibited hypomethylation at 3′ CpG islands within the Phospho1 gene, potentially regulating gene expression. (142) One system within which PHOSPHO1 plays a significant role in this regard is erythropoiesis. An expression quantitative trait locus (eQTL) analysis initially found that single-nucleotide polymorphisms (SNPs) associated with β-thalassemia in human peripheral blood samples were also associated with changes in PHOSPHO1 expression.…”