Canthin-6-One Accelerates Alpha-Synuclein Degradation by Enhancing UPS Activity: Drug Target Identification by CRISPR-Cas9 Whole Genome-Wide Screening Technology
Canthin-6-One Accelerates Alpha-Synuclein Degradation by Enhancing UPS Activity: Drug Target Identification by CRISPR-Cas9 Whole Genome-Wide Screening Technology
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DOI：
10.3389/fphar.2019.00016
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发表时间：
2019-01
影响因子：
5.6
通讯作者：
Ning-Ning Yuan-Ning;Cui-Zan Cai;Ming-Yue Wu;Qi Zhu;Huanxing Su;Min Li;Jiaoyan Ren;Jieqiong Tan;Jiahong Lu
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文献类型：
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作者：
Ning-Ning Yuan-Ning;Cui-Zan Cai;Ming-Yue Wu;Qi Zhu;Huanxing Su;Min Li;Jiaoyan Ren;Jieqiong Tan;Jiahong Lu
帕金森病 (PD) 是第二常见的神经退行性疾病，其特征是大脑黑质区域的多巴胺能神经元中蛋白质聚集体（即路易体）的积累。 α-突触核蛋白 (α-syn) 是 PD 患者路易体的主要成分，泛素-蛋白酶体系统的损伤与其积累有关。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 α-syn-EGFP 和亮红色荧光蛋白标记物 (mCherry) 的表达，以筛选潜在的降解 α-syn 的化合物。我们确定canthin-6-one是一种α-syn降低化合物，它以泛素蛋白酶体系统（UPS）依赖的方式促进野生型和突变体α-syn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作为canthin-6-one药理活性的靶基因。最后，我们发现canthin-6-one通过激活PKA上调PSMD1并增强UPS功能。
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 substantia nigra 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.