Gut-muscle axis: establishing the connection between microbial signaling and skeletal muscle function in a mouse model (mdx) of Duchenne muscular dystrophy

Abstract

The metabolic and endocrine nature of skeletal muscle allows for systemic signaling and influence over the enteric environment influencing gut microbial populations. Concomitantly, commensal microbial signaling can alter the composition and development of skeletal muscle forming a bidirectional gut-muscle axis. Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disease characterised by skeletal muscle atrophy, fibrosis, and ultimately cardiorespiratory failure. Previous studies have shown that gut microbial signaling can alter the pathways involved in muscle atrophy, enhancing atrophy when dysbiotic, or blunting the atrophic effect by restoring gut microbial populations to a healthy status; thus, we sought to characterize the metabolic signaling between the gut and the diaphragm in a mouse model of DMD ( mdx) to identify potential therapeutic targets. The study was divided into two arms and conducted following institutional ethical approval. The first arm investigated metabolomic signalling. 6-month-old male BL10 wildtype (WT; n = 12) and mdx (n = 12) mice were anaesthetized (induction by 5% isoflurane) and decapitated. Following collection of trunk blood, the diaphragm was excised, and colonic/cecal contents were collected. The samples were then processed by mass-spectrometry to isolate the metabolomic profiles of semi-polar, secondary bile acid, and short chain fatty acid (SCFA) targets in the gut, plasma and diaphragm of WT and mdx mice. The second arm of the study explored the gut microbial profile and diaphragm gene expression, structure and function. 6-month-old male WT (n = 15) and mdx (n = 15) were anesthetized (induction by 5% isoflurane) followed by cervical dislocation. The diaphragm was excised, and longitudinal strips were isolated for structural analysis (i.e., histological staining and immunohistochemistry for fibre type and size), PCR gene expression, and isometric force assessment ex vivo. Additionally, faecal pellets were collected and analysed via shotgun metagenomic sequencing to assess microbial populations. Diaphragm twitch and tetanic force in mdx were profoundly decreased compared to WT. Metabolic analysis of faecal metabolites indicated impairment in the three main tryptophan metabolic pathways in mdx mice: kynurenine, indole and serotonin. Metagenomic analysis indicated a profound genotype effect between WT and mdx gut microbial populations. Two main producers of the short chain fatty acid acetate (Lactobacillus and Bifidobacterium) were significantly decreased within the mdx group. Acetate serves as a substate in the TCA cycle contributing to ATP production and positively affects skeletal muscle via numerous pathways including the down regulation of key atrogenes associated with atrophy. Pathway enrichment analysis showed a global disruption to both purine and pyrimidine metabolism in all three tissue types. Purine/pyrimidine metabolism have been tightly linked with microbial metabolism and affect skeletal muscle health and function. Further understanding of the gut-muscle axis in dystrophic models may pave the way for novel microbiota-directed therapeutic strategies to ameliorate muscle dysfunction in DMD. Funded by Science Foundation Ireland SFI/19/6628 INSPIRE DMD. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.