The role of DNA Methylation in Anderson Fabry Disease pathogenesis: novel nutrigenomic frontier in a CRISPR/cas9-based cellular model

Anderson Fabry disease (AFD) is a rare X-linked lysosomal storage disorder caused by partial or total deficiency of the enzyme α-galactosidase A (α-GAL A), resulting in the accumulation of globotriaosylceramide (GB3) in lysosomes. Although research has mainly focused on genetic mutations of the GLA gene, growing evidence suggests that epigenetic mechanisms, particularly DNA methylation, may also contribute to disease heterogeneity, especially among female carriers.This study investigated the role of DNA methylation in AFD pathogenesis by analyzing global and gene-specific methylation profiles in peripheral blood mononuclear cells (PBMCs) from female AFD patients and healthy controls. The results revealed significant global DNA hypomethylation in AFD patients, indicative of genomic instability and chronic inflammation, together with hypermethylation of the GLA promoter region, potentially responsible for further gene silencing and reduced α-GAL A expression. These findings highlight a dual mechanism, genetic and epigenetic, underlying disease severity and variability.To explore these mechanisms in vitro, a CRISPR/Cas9-engineered cellular model of AFD, called HEK-GLAKD, was developed, reproducing the characteristic hypomethylated and inflammatory profile of the disease. This model was subsequently used to assess the effects of plant-derived extracellular vesicles (FicoVes), previously reported to possess anti-inflammatory properties. Treatment with FicoVes restored global methylation levels, reduced Interleukin-6 (IL-6) and Interleukin-8 (IL-8) gene expression and secretion and decreased lipid accumulation, indicating an overall anti-inflammatory and epigenetically stabilizing effect. Importantly, these findings suggest that FicoVes may represent a valuable adjuvant co-treatment to current enzyme replacement therapy (ERT), acting through complementary epigenetic and anti-inflammatory pathways. Collectively, this work demonstrates that DNA methylation plays a critical role in AFD pathophysiology and highlights the therapeutic potential of epigenetic modulation. Moreover, the established AFD cellular model provides a robust platform for future mechanistic studies and for the development of innovative adjuvant strategies targeting both genetic and epigenetic aspects of the disease. Moreover, by modulating DNA methylation and cytokine expression, FicoVes could enhance the efficacy of existing therapeutic approaches and contribute to improved clinical outcomes in AFD patients.