Restoration of Lysosomal Hydrolase Activities by LRRK2 Inhibition in GBA1 ‐ and LRRK2 ‐Associated Parkinson's Disease Patient‐Derived Cells

作者：Tatiana Usenko, Katerina Basharova, Anastasia Bezrukova, Е. В. Григорьева, С. В. Павлова, Galina Baydakova, Михаил Николаев, Ivan Gaponenko, O. V. Khegay, Irina Miliukhina, Ekaterina Zakharova, С. Н. Пчелина · 发表于：Journal of Neurochemistry · 年份：2025 · DOI：10.1111/jnc.70214 · 被引用次数：2 · 研究领域：Lysosomal Storage Disorders Research、Parkinson's Disease Mechanisms and Treatments、Autophagy in Disease and Therapy

Mutations in the GBA1 gene, encoding the lysosomal enzyme glucocerebrosidase (GCase), and the LRRK2 gene, encoding leucine-rich repeat kinase 2 (LRRK2) are the most common genetic risk factors for Parkinson's disease (PD). The potential use of LRRK2 inhibitors for treating not only LRRK2-associated PD (LRRK2-PD) but also GBA1-associated PD (GBA1-PD) is currently under discussion. In the present study, we aimed to evaluate whether LRRK2 inhibition affects lysosomal hydrolase enzymatic activities, autophagy, and alpha-synuclein levels in various cell types derived from LRRK2-PD and GBA1-PD patients, including macrophages derived from peripheral blood mononuclear cells (PBMC-derived macrophages), dopaminergic (DA) neurons derived from induced pluripotent stem cells (iPSC-derived DA neurons), and SH-SY5Y cells. We first characterized PBMC-derived macrophages from patients with LRRK2-PD. Confocal microscopy revealed reduced colocalization of GCase with the lysosomal marker LAMP2, indicating impaired lysosomal translocation of the enzyme. In parallel, we observed decreased activity of galactosylceramidase (GALC), increased levels of lysosphingolipids (HexSph, LysoGb3, LysoSM) as measured by LC-MS/MS, and decreased pro-form of cathepsin D, as assessed by Western blot analysis, compared to controls. However, no significant differences in GCase activity and protein levels were observed in PBMC-derived macrophages of LRRK2-PD patients and controls. In subsequent experiments, we found t...