Proteomic Profiling to Investigate the Effects of HDAC8 Inhibition Combined to the Activation of Sirtuin1 in a Zebrafish Model of Duchenne Muscular Dystrophy

Duchenne Muscular Dystrophy (DMD) is an X-linked degenerative genetic disorder caused by mutations of the DMD gene encoding dystrophin protein. DMD has an incidence of 1 in 3,500-5,000 live male births and a prevalence of 7.1 in 100,000 males worldwide. It is characterized by progressive skeletal muscle degeneration, inflammation, and fibrosis. While gene- and cell-mediated therapies hold promise, significant hurdleslimit their widespread application. Consequently, pharmacologicalstrategies aimed at reducing muscle loss and fibrosis and mitigating inflammation. Among the pharmacological candidates, histone deacetylase modulators (HDACi) have emerged as potential treatments. Specifically, HDAC8 plays a role in cytoskeletal remodelling, while Sirtuin1 (SIRT1) addresses mitochondrial biogenesis and energy metabolism. In this work, we demonstrated that the combination of HDAC8 inhibition with SIRT1 activation synergistically improves muscle structure and function and limits inflammation. In addition, in combined treatments it is possible to scale down the doses of a single drug, reducing side effects. To accomplish this, we took advantage of the dmd zebrafish model, and we compared the effects on skeletal muscle recovery and induction of anti-inflammatory, antifibrotic, and antioxidant effects. We analysed the acetylation profile of HDAC8 inhibited and SIRT1 activated zebrafish embryos to identify the respective acetylation targets. The identification of common and different targets of HDAC8 and SIRT1 from the acetylation profile holds significant potential for advancing therapeutic strategies explaining the mechanism of actions of the two compounds