Curcumin controls lysosomal acidification and exocytosis to ameliorate lysosomal cholesterol accumulation in Niemann–Pick type C disease

BACKGROUND AND PURPOSE: levels, and enhances lysosomal activation via mTOR suppression and TFEB activation. How CUR specifically targets lysosomal cholesterol has not been fully clarified. EXPERIMENTAL APPROACH: Effects of curcumin on lysosomal cholesterol accumulation were evaluated in NPC1 cell models. Filipin staining, immunofluorescence, surface LAMP1 and NAGase activity and Cathepsin B activity were investigated to evaluate lysosomal cholesterol, TFEB translocation, lysosomal exocytosis and hydrolytic activity. Lysosomal acidification was determined by Lysotracker Red, Oregon Green, and sfGFP/mCherry transfection. CRISPR/Cas9 and siRNA interference were used to investigate the role of TFEB/TFE3 and TRPML1 in curcumin-induced cholesterol reduction. KEY RESULTS: CUR alleviates lysosomal cholesterol accumulation in NPC1 cells in a TFEB- and TFE3-dependent manner. CUR enhanced lysosomal acidity and promoted calcium-dependent lysosomal exocytosis, which contributed to CUR-mediated lysosomal cholesterol clearance. The combination of CUR with specific agonists (ML-SAs) of MCOLN1/TRPML1, a lysosomal cation channel required for lysosomal exocytosis, improved lysosomal cholesterol clearance. CONCLUSION AND IMPLICATIONS: - and TRPML1-dependent lysosomal exocytosis. These findings support curcumin and its analogues as potential therapeutics for NPCD and other diseases of lysosomal storage.