Disrupting the Feed-Forward Cycle of RyR1 Calcium Leak and Oxidative Stress Mitigates Doxorubicin-Induced Skeletal Myopathy

Abstract

Doxorubicin (DOX), a highly effective and widely used chemotherapeutic agent used to treat various types of cancer. Unfortunately, DOX also has some undesirable and off-target effects, particularly debilitating muscle weakness and fatigue. The mechanism behind this DOX-induced skeletal myotoxicity (DISM) remains unclear. Here, we show that acute DOX exposure, at clinically relevant concentrations, impairs isometric force production and accelerates fatigue in ex vivo murine flexor digitorum brevis (FDB) muscles. Mechanistically, we found that DOX increases the open probability of single RyR1 and disrupts calcium (Ca2+)-dependent inactivation (CDI). This results in a persistent sarcoplasmic reticulum (SR) Ca2+ leak, elevated basal cytosolic Ca2+, and abnormal Ca2+ release during action potentials. This abnormal intracellular Ca2+ handling ultimately leads to increased mitochondrial reactive oxygen species (ROS) production, which, in turn, exacerbates the functional instability of RyR1. Interestingly, the cytosolic basal Ca2+ elevation precedes ROS generation, suggesting that it initiates a destructive cross-talk between Ca2+ dysregulation and oxidative stress. Notably, pharmacological stabilization of the RyR1-FKBP12 complex with novel triazole compounds, MP-001 and MP-034, normalizes RyR1 function, Ca2+ and ROS homeostasis, as well as muscle force and fatigue resistance. Our findings indicate that DISM is initiated by DOX destabilization of the RyR1-FKBP12 complex (abnormal SR Ca2+ leak) and then exacerbated by the Ca-ROS vicious cycle. Limiting RyR1-mediated Ca2+ leak with MP-001 represents a promising therapeutic strategy for anti-DISM, aiming to normalize muscle function in patients undergoing DOX chemotherapy.