Polymeric nanoparticles delivery of AMPK activator 991 prevents its toxicity and improves muscle homeostasis in Duchenne Muscular Dystrophy

Abstract

AbstractMuscular dystrophies, such as Duchenne muscular dystrophy (DMD), are caused by permanent muscle injuries leading to chronic inflammation. In that context, macrophages harbor an altered inflammatory profile that contributes to fibrosis through the secretion of the profibrotic cytokine TGFβ1. We previously showed that AMP-activated protein kinase (AMPK) activation reduces TGFβ1 secretion by macrophages and improves muscle homeostasis and muscle force in a mouse model of DMD. This makes AMPK an attractive therapeutic target for treating chronic inflammation and fibrosis in DMD. However, potent direct AMPK activators like compound 991 show strong adverse effectsin vivo,preventing their direct use. Here, we encapsulated 991 into biodegradable polymeric poly(lactic-co-glycolic) acid (PLGA) nanoparticles forin vivodelivery, in an attempt to overcome toxicity issues. We show that 991-loaded PLGA nanoparticles retained drug activity on fibrotic macrophagesin vitro, by reducing their secretion of TGFβ1. In the D2-mdx pre-clinical DMD mouse model, intravenously injected PLGA nanoparticles reached gastrocnemius and diaphragm muscles, which are the most affected muscles in this model. Chronic intravenous injections of 991-loaded PLGA nanoparticles decreased inflammation in both muscles, which was associated with fibrosis reduction and increase in myofiber size and muscle mass in the gastrocnemius. No impact on blood cell counts and liver enzymes was observed. These results demonstrate that nanomedicine is an efficient strategy to deliver AMPK activatorsin vivoto target inflammation and improve the dystrophic muscle phenotype.