OMA1 High-Throughput Screen Reveals Protease Activation by Kinase Inhibitors,ACS Chemical Biology

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OMA1 High-Throughput Screen Reveals Protease Activation by Kinase Inhibitors
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2021-10-21 , DOI: 10.1021/acschembio.1c00350
Marcel V Alavi ₁

Affiliation

Mitochondrial proteases are interesting but challenging drug targets for multifactorial diseases, such as neurodegeneration and cancer. The mitochondrial inner membrane protease OMA1 is a bona fide drug target for heart failure supported by data from human linkage analysis and animal disease models, but presumably relevant for more indications. OMA1 acts at the intersection of energy metabolism and stress signaling. The protease cleaves the structural protein OPA1, which organizes the cristae, as well as the signaling peptide DELE1, which can stimulate the integrated stress response. OMA1 shows little activity under physiological conditions but hydrolyzes OPA1 in mitochondria destined for mitophagy and during apoptosis. Little is known about OMA1, its structure has not been solved, let alone its context-dependent regulation. Autocatalytic processing and the lack of OMA1 inhibitors are thereby creating the biggest roadblocks. This study introduces a scalable, cellular OMA1 protease assay suitable for high-throughput drug screening. The assay utilizes an engineered luciferase targeted to the inner membrane as artificial OMA1 substrate, whereby the reporter signal inversely correlates to OMA1 activity. Testing different screening protocols and sampling different compound collections validated the reporter and demonstrated that both OMA1 activators as well as OMA1 inhibitors can be identified with the assay. Ten kinase-targeted cancer drugs triggered OMA1 in the assays, which suggests─considering cardiotoxicity as a rather common side-effect of this class of drugs─cross-reactivity with the OMA1 pathway.

中文翻译：

OMA1 高通量筛选揭示了激酶抑制剂对蛋白酶的激活

线粒体蛋白酶是有趣但具有挑战性的多因素疾病药物靶点，例如神经退行性疾病和癌症。线粒体内膜蛋白酶 OMA1 是真正的心力衰竭药物靶标，得到人类连锁分析和动物疾病模型数据的支持，但可能与更多适应症相关。OMA1 作用于能量代谢和应激信号的交叉点。蛋白酶切割组织嵴的结构蛋白 OPA1，以及可以刺激综合应激反应的信号肽 DELE1。OMA1 在生理条件下几乎没有活性，但会水解线粒体中用于线粒体自噬和细胞凋亡过程中的 OPA1。关于 OMA1 知之甚少，其结构尚未解决，更不用说其上下文相关的调控了。因此，自催化加工和缺乏 OMA1 抑制剂是最大的障碍。本研究介绍了一种适用于高通量药物筛选的可扩展细胞 OMA1 蛋白酶测定。该测定利用靶向内膜的工程荧光素酶作为人工 OMA1 底物，报告信号与 OMA1 活性呈负相关。测试不同的筛选方案并对不同的化合物集合进行取样验证了报告者，并证明了 OMA1 激活剂和 OMA1 抑制剂都可以通过该检测进行鉴定。十种靶向激酶的抗癌药物在试验中触发了 OMA1，这表明──将心脏毒性视为此类药物相当常见的副作用──与 OMA1 通路存在交叉反应。

更新日期：2021-11-19

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