Bezrukova A I, Basharova K S, Emelyanov A K, Rybakov A V, Miliukhina I V, Pchelina S N, Usenko T S
Petersburg Nuclear Physics Institute named by B.P.Konstantinov of NRC «Kurchatov Institute», 1, mkr. Orlova roshcha, 188300, Gatchina, Russia.
Pavlov First Saint Petersburg State Medical University, 6-8 Lva Tolstogo Street, 197022, Saint Petersburg, Russia.
Biochem Genet. 2025 May 19. doi: 10.1007/s10528-025-11125-z.
Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the loss of dopaminergic neurons and abnormal aggregation of the alpha-synuclein protein. Disruption of the autophagy-lysosomal pathway is closely associated with PD pathogenesis. Here, using western-blot analysis we assessed the level of autophagy-related proteins, including phosphorylated mTOR (p-mTOR), phosphorylated RPS6 (p-RPS6), beclin-1 (BECN1), LC3B, p62, and cathepsin D (CTSD) in macrophages derived from peripheral blood mononuclear cells (PBMC-derived macrophages) of GBA1-PD (p.N370S/N, p.L444P/N), LRRK2-PD (p.G2019S/N), idiopathic PD (iPD) patients, and healthy controls. Our findings revealed mutation-specific disruptions in autophagy pathways among PD patients. In p.N370S-GBA1-PD, PBMC-derived macrophages exhibited elevated levels of p-RPS6, BECN1, LC3B-II and decreased mature form of CTSD levels suggesting more active mTOR-dependent autophagy initiation alongside potential autophagosome accumulation that may lead to downregulation of lysosomal degradation. p.L444P-GBA1-PD PBMC-derived macrophages showed increased levels of p-RPS6 and BECN1, coupled with decreased p62 levels and stable mature form of CTSD and LC3B-II, indicative of enhanced autophagy flux driven by mTOR activity without evident lysosomal dysfunction. In p.G2019S-LRRK2-PD patients, PBMC-derived macrophages demonstrated elevated p-RPS6, LC3B-II, and mature CTSD levels, alongside reduced p62 levels. These changes suggest higher basal autophagosome abundance in steady-state autophagy and turnover, potentially driven by lysosomal alterations rather than direct mTOR dysregulation. These mutation-dependent differences highlight distinct autophagy dynamics in GBA1-PD and LRRK2-PD, underscoring the critical role of genetic mutations in modulating PD pathogenesis. Our results emphasize the necessity for subtype-specific therapeutic strategies targeting autophagy and other mTOR-regulated pathways to address the heterogeneity of PD mechanisms.
帕金森病（PD）是一种常见的神经退行性疾病，其特征是多巴胺能神经元丧失以及α-突触核蛋白异常聚集。自噬-溶酶体途径的破坏与PD发病机制密切相关。在此，我们使用蛋白质免疫印迹分析评估了来自GBA1-PD（p.N370S/N、p.L444P/N）、LRRK2-PD（p.G2019S/N）、特发性PD（iPD）患者及健康对照者外周血单核细胞来源的巨噬细胞（PBMC来源的巨噬细胞）中自噬相关蛋白的水平，这些蛋白包括磷酸化的mTOR（p-mTOR）、磷酸化的RPS6（p-RPS6）、贝林1（BECN1）、LC3B、p62和组织蛋白酶D（CTSD）。我们的研究结果揭示了PD患者自噬途径中特定突变导致的破坏。在p.N370S-GBA1-PD中，PBMC来源的巨噬细胞表现出p-RPS6、BECN1、LC3B-II水平升高，而成熟形式的CTSD水平降低，这表明mTOR依赖性自噬起始更活跃，同时可能存在自噬体积累，这可能导致溶酶体降解下调。p.L444P-GBA1-PD的PBMC来源的巨噬细胞显示p-RPS6和BECN1水平升高，同时p62水平降低，CTSD和LC3B-II的成熟形式稳定，这表明由mTOR活性驱动的自噬通量增强，且无明显的溶酶体功能障碍。在p.G2019S-LRRK2-PD患者中，PBMC来源的巨噬细胞表现出p-RPS6、LC3B-II和成熟CTSD水平升高，同时p62水平降低。这些变化表明在稳态自噬和周转中基础自噬体丰度更高，这可能是由溶酶体改变而非直接的mTOR失调驱动的。这些依赖于突变的差异突出了GBA1-PD和LRRK2-PD中不同的自噬动态，强调了基因突变在调节PD发病机制中的关键作用。我们的结果强调了针对自噬和其他mTOR调节途径的亚型特异性治疗策略对于解决PD机制异质性的必要性。
