Accumulation of α-synuclein is a main underlying pathological feature of Parkinson's disease and α-synucleinopathies, for which lowering expression of the α-synuclein gene (SNCA) is a potential therapeutic avenue. Using a cell-based luciferase reporter of SNCA expression we performed a quantitative high-throughput screen of 155,885 compounds and identified A-443654, an inhibitor of the multiple functional kinase AKT, as a potent inhibitor of SNCA. HEK-293 cells with CAG repeat expanded ATXN2 (ATXN2-Q58 cells) have increased levels of α-synuclein. We found that A-443654 normalized levels of both SNCA mRNA and α-synuclein monomers and oligomers in ATXN2-Q58 cells. A-443654 also normalized levels of α-synuclein in fibroblasts and iPSC-derived dopaminergic neurons from a patient carrying a triplication of the SNCA gene. Analysis of autophagy and endoplasmic reticulum stress markers showed that A-443654 successfully prevented α-synuclein toxicity and restored cell function in ATXN2-Q58 cells, normalizing the levels of mTOR, LC3-II, p62, STAU1, BiP, and CHOP. A-443654 also decreased the expression of DCLK1, an inhibitor of α-synuclein lysosomal degradation. Our study identifies A-443654 and AKT inhibition as a potential strategy for reducing SNCA expression and treating Parkinson's disease pathology.

中文翻译：

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AKT 调节剂 A-443654 可降低 α-突触核蛋白表达并使 ER 应激和自噬正常化。

α-突触核蛋白的积累是帕金森病和 α-突触核蛋白病的主要潜在病理特征，为此降低 α-突触核蛋白基因 (SNCA) 的表达是一种潜在的治疗途径。使用基于细胞的 SNCA 表达荧光素酶报告基因，我们对 155,885 种化合物进行了定量高通量筛选，并鉴定出 A-443654（一种多功能激酶 AKT 抑制剂）是一种有效的 SNCA 抑制剂。具有 CAG 重复扩增 ATXN2 的 HEK-293 细胞（ATXN2-Q58 细胞）具有增加的 α-突触核蛋白水平。我们发现 A-443654 使 ATXN2-Q58 细胞中 SNCA mRNA 和 α-突触核蛋白单体和寡聚体的水平正常化。A-443654 还使来自携带三倍 SNCA 基因的患者的成纤维细胞和 iPSC 衍生的多巴胺能神经元中的 α-突触核蛋白水平正常化。自噬和内质网应激标记物分析表明，A-443654 成功阻止了 ATXN2-Q58 细胞中的 α-突触核蛋白毒性并恢复了细胞功能，使 mTOR、LC3-II、p62、STAU1、BiP 和 CHOP 水平正常化。A-443654 还降低了 DCLK1 的表达，DCLK1 是 α-突触核蛋白溶酶体降解的抑制剂。我们的研究确定 A-443654 和 AKT 抑制是降低 SNCA 表达和治疗帕金森病病理的潜在策略。