Petersburg Nuclear Physics Institute Named by B.P. Konstantinov of National Research Center «Kurchatov Institute», Gatchina 188300, Russia.
Department of Molecular Genetic and Nanobiological Technologies, Pavlov First Saint-Petersburg State Medical University, Saint-Petersburg 197022, Russia.
Int J Mol Sci. 2023 May 22;24(10):9105. doi: 10.3390/ijms24109105.
Mutations in the gene, encoding the lysosomal enzyme glucocerebrosidase (GCase), cause Gaucher disease (GD) and are the most common genetic risk factor for Parkinson's disease (PD). Pharmacological chaperones (PCs) are being developed as an alternative treatment approach for GD and PD. To date, NCGC00241607 (NCGC607) is one of the most promising PCs. Using molecular docking and molecular dynamics simulation we identified and characterized six allosteric binding sites on the GCase surface suitable for PCs. Two sites were energetically more preferable for NCGC607 and located nearby to the active site of the enzyme. We evaluated the effects of NCGC607 treatment on GCase activity and protein levels, glycolipids concentration in cultured macrophages from GD ( = 9) and GBA-PD ( = 5) patients as well as in induced human pluripotent stem cells (iPSC)-derived dopaminergic (DA) neurons from GBA-PD patient. The results showed that NCGC607 treatment increased GCase activity (by 1.3-fold) and protein levels (by 1.5-fold), decreased glycolipids concentration (by 4.0-fold) in cultured macrophages derived from GD patients and also enhanced GCase activity (by 1.5-fold) in cultured macrophages derived from GBA-PD patients with N370S mutation ( < 0.05). In iPSC-derived DA neurons from GBA-PD patients with N370S mutation NCGC607 treatment increased GCase activity and protein levels by 1.1-fold and 1.7-fold ( < 0.05). Thus, our results showed that NCGC607 could bind to allosteric sites on the GCase surface and confirmed its efficacy on cultured macrophages from GD and GBA-PD patients as well as on iPSC-derived DA neurons from GBA-PD patients.
基因突变 ，编码溶酶体酶葡萄糖脑苷脂酶（GCase），导致戈谢病（GD），是帕金森病（PD）最常见的遗传危险因素。药理学伴侣（PCs）被开发为 GD 和 PD 的替代治疗方法。迄今为止，NCGC00241607（NCGC607）是最有前途的 PCs 之一。我们使用分子对接和分子动力学模拟在 GCase 表面上鉴定并表征了六个适合 PCs 的别构结合位点。两个位点在能量上更适合 NCGC607，并且位于酶的活性位点附近。我们评估了 NCGC607 治疗对 GD（=9）和 GBA-PD（=5）患者培养的巨噬细胞以及 GBA-PD 患者诱导的人多能干细胞（iPSC）衍生的多巴胺能（DA）神经元中 GCase 活性和蛋白水平、糖脂浓度的影响。结果表明，NCGC607 治疗增加了 GD 患者来源的巨噬细胞中的 GCase 活性（增加了 1.3 倍）和蛋白水平（增加了 1.5 倍），降低了糖脂浓度（降低了 4.0 倍），并且还增强了 GBA-PD 患者来源的巨噬细胞中的 GCase 活性（增加了 1.5 倍）与 N370S 突变（ < 0.05）。在 GBA-PD 患者中，NCGC607 治疗增加了 N370S 突变的 iPSC 衍生 DA 神经元中的 GCase 活性和蛋白水平，分别增加了 1.1 倍和 1.7 倍（ < 0.05）。因此，我们的结果表明，NCGC607 可以与 GCase 表面的别构位点结合，并证实其对 GD 和 GBA-PD 患者培养的巨噬细胞以及 GBA-PD 患者 iPSC 衍生的 DA 神经元均有效。
