Lysosomal Ca2+ as a mediator of palmitate-induced lipotoxicity

Abstract

While the mechanism of lipotoxicity by palmitic acid (PA), an effector of metabolic stress in vitro and in vivo, has been extensively investigated, molecular details of lipotoxicity are still not fully characterized. Since recent studies reported that PA can exert lysosomal stress in addition to well-known ER and mitochondrial stress, we studied the role of lysosomal events in lipotoxicity by PA, focusing on lysosomal Ca²⁺. We found that PA induced accumulation of mitochondrial ROS and that mitochondrial ROS induced release of lysosomal Ca²⁺ due to lysosomal Ca²⁺ exit channel activation. Lysosomal Ca²⁺ release led to increased cytosolic Ca²⁺ which induced mitochondrial permeability transition (mPT). Chelation of cytoplasmic Ca²⁺ or blockade of mPT with olesoxime or decylubiquinone (DUB) suppressed lipotoxicity. Lysosomal Ca²⁺ release led to reduced lysosomal Ca²⁺ content which was replenished by ER Ca²⁺, the largest intracellular Ca²⁺ reservoir (ER → lysosome Ca²⁺ refilling), which in turn activated store-operated Ca²⁺ entry (SOCE). Inhibition of ER → lysosome Ca²⁺ refilling by blockade of ER Ca²⁺ exit channel using dantrolene or inhibition of SOCE using BTP2 inhibited lipotoxicity in vitro. Dantrolene or DUB also inhibited lipotoxic death of hepatocytes in vivo induced by administration of ethyl palmitate together with LPS. These results suggest a novel pathway of lipotoxicity characterized by mPT due to lysosomal Ca²⁺ release which was supplemented by ER → lysosome Ca²⁺ refilling and subsequent SOCE, and also suggest the potential role of modulation of ER → lysosome Ca²⁺ refilling by dantrolene or other blockers of ER Ca²⁺ exit channels in disease conditions characterized by lipotoxicity such as metabolic syndrome, diabetes, cardiomyopathy or nonalcoholic steatohepatitis.