Effects of methyl donor nutrient supplementation on DNA methylation profile in patients with lupus and normal weight or excess body weight: a randomized clinical study

Author(s)

Jhulia Caroline Nunes Leal da Mota

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD)
• Defense date: 2024-06-25

Examining board members

Carolina Nicoletti Ferreira Fino; Fabiana Braga Benatti; Maria Aderuza Horst; Lígia Moriguchi Watanabe

• Advisor: Carolina Nicoletti Ferreira Fino

Abstract

Systemic lupus erythematosus (SLE) is a multifaceted chronic autoimmune condition characterized by a complex interplay of genetic, epigenetic, immunological, and environmental factors. Alterations in epigenetic patterns have been implicated in the development of autoimmunity and specific SLE´s phenotypic characteristics. Nutrients such as folic acid and vitamin B12 act as cofactors or methyl group donors, influencing DNA methylation profiles. In this double-blind controlled clinical trial, we aimed to investigate whether supplementation with folic acid and vitamin B12 could modulate the DNA methylation profile in the adipose tissue of SLE patients with either normal weight or excess body weight, and whether this modulation differed according to nutritional status. Fifty-one premenopausal women aged 18 to 45 years, in disease remission, receiving prednisone <10 mg/day and stable hydroxychloroquine dose, were divided into two groups based on BMI. The normal weight group comprised patients with adequate weight (age: 33.7±7.1 years; BMI: 22.1±2.4 kg/m²), while the excess body weight group included those with excess weight (age: 36.3±5.5 years; BMI: 30.4±3.6 kg/m²). Each group was randomized to receive either vitamin mix supplementation (2.000 mcg of vitamin B12 with 400 mcg of folic acid) or placebo. Clinical, anthropometric, metabolic, and dietary parameters were evaluated before and after 3 months of intervention, with samples of peripheral blood and subcutaneous adipose tissue collected. DNA methylation analysis was conducted using the Infinium Human Methylation EPIC Beadchip platform, and gene expression analysis was performed using real-time quantitative polymerase chain reaction by 2-Ct method. Both were performed with subcutaneous adipose tissue samples. Shapiro-Wilk, t-test for independent samples and chi-square test were used for baseline analyses, while the Generalized Estimating Equation (with Bonferroni post hoc) was used to analyze the effect of the intervention (p<0.05). Changes in methylation levels of CpGs were assessed, considering a minimum of 5%, p<0.001, and false discovery rate <0.05. While serum concentrations of folic acid and vitamin B12 increased post-supplementation in both groups, no significant alterations in phenotypic characteristics were noted. In patients with normal weight, 418 differentially methylated CpGs were identified post-supplementation, associated with metabolic pathways regulating NODtype receptors (NLRs) signaling, xenobiotic metabolism via cytochrome P450, and carcinogenesis pathways. Conversely, patients with excess body weight showed 657 differentially methylated CpGs, linked to tumor necrosis factor (TNF)- signaling, insulin regulation, and oxidative phosphorylation. Notably, there were no changes in gene expression of TNF-, IL-6, MTHFR, DNMT1, STAT3, ADIPOQ, and LEP following supplementation in either group. These findings suggest that methyl group donor micronutrients modulate adipose tissue DNA methylation profiles in women with SLE, and this effect of supplementation is different depending on the nutritional status. This study was registered at clinicaltrials.gov (nº.: NCT05097365) (AU)

• FAPESP's process: 21/09745-5 - Effect of methyl-donor nutrient supplementation on methylation profile of inflammatory-related genes in lupus patients with obesity: a clinical trial
• Grantee: Jhulia Caroline Nunes Leal da Mota
• Support Opportunities: Scholarships in Brazil - Master