Abstract

A critical question in Duchenne muscular dystrophy (DMD) research is whether regulatory mechanisms beyond dystrophin loss contribute to impaired muscle regeneration. Through integrative analysis of proteomics and single-nucleus RNA sequencing database, we identify the upregulation of ANXA11 , a gene encoding a Ca²⁺-dependent phospholipid-binding protein, in MYH3⁺ regenerative myofibers from both mdx mice and DMD patients. This upregulation disrupts the maturation of regenerative myofibers, preventing adequate compensation for muscle loss in mdx mice due to dysregulation of the mTOR pathway. Suppression of Anxa11 via genetic knockout or AAV9-mediated knockdown significantly enhanced MYH3⁺ myofiber maturation, accompanied by restored S6 phosphorylation and robust functional muscle recovery in mdx mice. These results establish ANXA11 as a key regulator of muscle regeneration and a potential therapeutic target for DMD.