Yuan Ning-Ning, Cai Cui-Zan, Wu Ming-Yue, Zhu Qi, Su HuanXing, Li Min, Ren JiaoYan, Tan Jie-Qiong, Lu Jia-Hong
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.
Mr. and Mrs. Ko Chi Ming Centre for Parkinson's Disease Research, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China.
Front Pharmacol. 2019 Jan 28;10:16. doi: 10.3389/fphar.2019.00016. eCollection 2019.
Parkinson's disease (PD) is the second most common neurodegenerative disorder characterized by the accumulation of protein aggregates (namely Lewy bodies) in dopaminergic neurons in the region of the brain. Alpha-synuclein (α-syn) is the major component of Lewy bodies in PD patients, and impairment of the ubiquitin-proteasome system has been linked to its accumulation. In this work, we developed a tetracycline-inducible expression system, with simultaneous induced expression of α-syn-EGFP and a bright red fluorescent protein marker (mCherry) to screen for potential compounds for degrading α-syn. We identified canthin-6-one as an α-syn lowering compound which promoted both wild type and mutants α-syn degradation in an ubiquitin-proteasome-system (UPS) dependent manner. By CRISPR/Cas9 genome-wide screening technology, we identified RPN2/PSMD1, the 26S proteasome non-ATPase regulatory subunit 1, as the targeting gene for pharmacological activity of canthin-6-one. Finally, we showed that canthin-6-one up-regulates PSMD1 and enhances UPS function by activating PKA.
帕金森病（PD）是第二常见的神经退行性疾病，其特征是大脑区域多巴胺能神经元中蛋白质聚集体（即路易小体）的积累。α-突触核蛋白（α-syn）是帕金森病患者路易小体的主要成分，泛素-蛋白酶体系统的损伤与它的积累有关。在这项工作中，我们开发了一种四环素诱导表达系统，同时诱导表达α-syn-EGFP和一种亮红色荧光蛋白标记物（mCherry），以筛选降解α-syn的潜在化合物。我们鉴定出6-酮基-canthin为一种降低α-syn的化合物，它以泛素-蛋白酶体系统（UPS）依赖的方式促进野生型和突变型α-syn的降解。通过CRISPR/Cas9全基因组筛选技术，我们鉴定出26S蛋白酶体非ATP酶调节亚基1（RPN2/PSMD1）为6-酮基-canthin药理活性的靶向基因。最后，我们表明6-酮基-canthin通过激活蛋白激酶A（PKA）上调PSMD1并增强UPS功能。
