Scientific Institute IRCCS E. Medea, Laboratory of Molecular Biology, 23842 Bosisio Parini, Lecco, Italy.
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Largo E. Meneghetti 2, Padova, Italy.
Brain. 2023 Mar 1;146(3):1103-1120. doi: 10.1093/brain/awac308.
SPG15 is a hereditary spastic paraplegia subtype caused by mutations in Spastizin, a protein encoded by the ZFYVE26 gene. Spastizin is involved in autophagosome maturation and autophagic lysosome reformation and SPG15-related mutations lead to autophagic lysosome reformation defects with lysosome enlargement, free lysosome depletion and autophagosome accumulation. Symptomatic and rehabilitative treatments are the only therapy currently available for patients. Here, we targeted autophagy and lysosomes in SPG15 patient-derived cells by using a library of autophagy-modulating compounds. We identified a rose of compounds affecting intracellular calcium levels, the calcium-calpain pathway or lysosomal functions, which reduced autophagosome accumulation. The six most effective compounds were tested in vivo in a new SPG15 loss of function Drosophila model that mimicked the reported SPG15 phenotype, with autophagosome accumulation, enlarged lysosomes, reduced free lysosomes, autophagic lysosome reformation defects and locomotor deficit. These compounds, namely verapamil, Bay K8644, 2',5'-dideoxyadenosine, trehalose, Small-Molecule Enhancer of Rapamycin 28 and trifluoperazine, improved lysosome biogenesis and function in vivo, demonstrating that lysosomes are a key pharmacological target to rescue SPG15 phenotype. Among the others, the Small-Molecule Enhancer of Rapamycin 28 was the most effective, rescuing both autophagic lysosome reformation defects and locomotor deficit, and could be considered as a potential therapeutic compound for this hereditary spastic paraplegia subtype.
SPG15 是一种遗传性痉挛性截瘫亚型,由 Spastizin 基因突变引起,该蛋白由 ZFYVE26 基因编码。Spastizin 参与自噬体成熟和自噬溶酶体再形成,SPG15 相关突变导致自噬溶酶体再形成缺陷,溶酶体增大、游离溶酶体耗竭和自噬体积累。目前,患者仅可接受对症和康复治疗。在这里,我们通过使用自噬调节化合物文库靶向 SPG15 患者来源细胞中的自噬和溶酶体。我们鉴定出了一组影响细胞内钙水平、钙钙蛋白酶途径或溶酶体功能的化合物,这些化合物可减少自噬体的积累。六种最有效的化合物在一种新的 SPG15 功能丧失的果蝇模型中进行了体内测试,该模型模拟了报道的 SPG15 表型,表现为自噬体积累、溶酶体增大、游离溶酶体减少、自噬溶酶体再形成缺陷和运动缺陷。这些化合物,即维拉帕米、Bay K8644、2',5'-二脱氧腺苷、海藻糖、雷帕霉素小分子增强剂 28 和三氟拉嗪,改善了体内溶酶体生物发生和功能,证明溶酶体是挽救 SPG15 表型的关键药物靶点。在其他化合物中,雷帕霉素小分子增强剂 28 最为有效,可同时挽救自噬溶酶体再形成缺陷和运动缺陷,可被认为是这种遗传性痉挛性截瘫亚型的潜在治疗化合物。