Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92122.
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92122.
Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2320257121. doi: 10.1073/pnas.2320257121. Epub 2024 Aug 16.
Lysosomal degradation pathways coordinate the clearance of superfluous and damaged cellular components. Compromised lysosomal degradation is a hallmark of many degenerative diseases, including lysosomal storage diseases (LSDs), which are caused by loss-of-function mutations within both alleles of a lysosomal hydrolase, leading to lysosomal substrate accumulation. Gaucher's disease, characterized by <15% of normal glucocerebrosidase function, is the most common LSD and is a prominent risk factor for developing Parkinson's disease. Here, we show that either of two structurally distinct small molecules that modulate PIKfyve activity, identified in a high-throughput cellular lipid droplet clearance screen, can improve glucocerebrosidase function in Gaucher patient-derived fibroblasts through an MiT/TFE transcription factor that promotes lysosomal gene translation. An integrated stress response (ISR) antagonist used in combination with a PIKfyve modulator further improves cellular glucocerebrosidase activity, likely because ISR signaling appears to also be slightly activated by treatment by either small molecule at the higher doses employed. This strategy of combining a PIKfyve modulator with an ISR inhibitor improves mutant lysosomal hydrolase function in cellular models of additional LSD.
溶酶体降解途径协调清除多余和受损的细胞成分。溶酶体降解功能受损是许多退行性疾病的标志,包括溶酶体贮积症(LSD),这是由溶酶体水解酶的两个等位基因中的功能丧失突变引起的,导致溶酶体底物积累。戈谢病的特征是葡萄糖脑苷脂酶功能正常的<15%,是最常见的 LSD,也是发展为帕金森病的一个显著危险因素。在这里,我们通过促进溶酶体基因翻译的 MiT/TFE 转录因子,表明在高通量细胞脂滴清除筛选中鉴定的两种结构上不同的、调节 PIKfyve 活性的小分子中的任何一种,都可以改善戈谢病患者来源的成纤维细胞中的葡萄糖脑苷脂酶功能。在与 PIKfyve 调节剂联合使用的整合应激反应(ISR)拮抗剂进一步提高了细胞葡萄糖脑苷脂酶活性,这可能是因为在较高剂量下,这两种小分子的治疗都能轻微激活 ISR 信号。这种将 PIKfyve 调节剂与 ISR 抑制剂相结合的策略,改善了其他 LSD 的细胞模型中突变型溶酶体水解酶的功能。