The anti-retroviral therapy emtricitabine affects skeletal muscle DNA methylation and transcriptome patterns in a male HIV mouse model

Abstract

BackgroundAlthough antiretroviral therapies (ART) have substantially reduced HIV-associated mortality, the increased lifespan achieved by widespread ART deployment has revealed that HIV infection is linked to an unexplained earlier onset and increased incidence of aging-associated conditions like sarcopenia. Complex syndromes, like sarcopenia, often arise from a combination of genetic and environmental factors, so in this study, we investigated effects of short-term treatment with emtricitabine (2,3-dideoxy-5-fluoro-3-thiacytidine; FTC), an FDA approved ART, on skeletal muscle DNA methylation patterns and transcriptome-wide responses in a male murine model of HIV phenotypic biology (Tg26 mice). MethodsWe treated 6 month old male Tg26 (+/-) mice or wildtype (WT) littermates on a C57BL/6 genetic background with FTC in the drinking water for one month; control groups received drinking water vehicle alone (VEH). Muscle function and body composition were measured longitudinally. Skeletal muscle methylation patterns, transcriptional changes, and histological features were quantified at sacrifice. ResultsAlthough neither gross structural nor functional muscle deficits were observed in this short-term study with ART usage, relative decreases in muscle endurance measured by hang time over the study were 2-fold more severe in the Tg26 as compared to WT mice (p=0.0453), and markers of myogenic cell maturation (Myf5, Myf6) were disrupted in the Tg26 HIV model as compared to WT littermates in a manner exacerbated by FTC treatment. Fat mass, measured by DXA, also tended (p=0.085) to be uniquely increased by FTC treatment in the Tg26 mice over the study. Differential methylation patterns and pathway enrichment data suggested that the presence of an HIV phenotype and exposure to an ART regimen altered the methylation status in skeletal muscle genes such as Camk2B, Pcolce2 and Lima1 in a manner consistent with promoting eventual functional impairment in muscle. Additionally, RNAseq revealed differential gene expression profiles and key regulatory pathways including cellular differentiation, regulation of lipid metabolism, and neuroactive ligand-receptor interactions. Lipodystrophy-related genes including LEP, ADIPOQ and PPAR involved in fat distribution and metabolism along with skeletal genes related to regulation of muscle strength were affected by the presence of an HIV phenotype and ART treatment. ConclusionsThe current study provides insights into mechanisms by which a clinically relevant ART may influence DNA methylation and transcriptome changes in skeletal muscle in the context of HIV biology. The differentially regulated pathways suggest novel targets for understanding, and eventually abrogating, the harmful effects of long-term ART use in PLWH on skeletal muscle mass and function.