CRISPR-activation screen identified potassium channels for protection against mycotoxins through cell cycle progression and mitochondrial function.

IF 4.1 Q2 CELL BIOLOGY
Yulong Tang, Simeng Liao, Zhuyuan Nie, Guangwei Kuang, Chunxiao Ji, Dan Wan, Liuqin He, Fengna Li, Xiangfeng Kong, Kai Zhan, Bie Tan, Xin Wu, Yulong Yin
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引用次数: 0

Abstract

Zearalenone (ZEA) exposure has carcinogenic effects on human and animal health by exhibiting intestinal, hepatic, and renal toxicity. At present, the underlying mechanisms on how ZEA induces apoptosis and damage to tissues still remain unclear. In this study, we aimed to identify genes that modulate the cellular response to ZEA using clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screening, and further validate novel gene functions to elucidate molecular mechanisms underlying particular biological processes in vivo and in vitro. Two ZEA-resistant cell lines, designated Ov-KCNJ4 and Ov-KCNJ12, were yielded by CRISPR activation screening which had significant changes in ZEA resistance and growth rates. Results showed that ZEA could interact with the cell membrane proteins KCNJ4 and KCNJ12, inducing cell cycle arrest, disruption of DNA replication and base excision repair. Overexpression of KCNJ4 and KCNJ12 was involved in ZEA resistance by regulating cell cycle to neutralize toxicity, sustaining mitochondrial morphology and function via attenuating the damage from oxidative stress in the KCNJ4-mitoKATP pathway. In vivo experiments showed that AAV-KCNJ4 delivery significantly improved ZEA-induced renal impairment and increased antioxidative enzyme activity by improving mitochondrial function. Our findings suggest that increasing potassium channel levels may be a putative therapeutic target for mycotoxin-induced damage.

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crispr激活筛选通过细胞周期进程和线粒体功能鉴定了防止霉菌毒素的钾通道。
玉米赤霉烯酮(ZEA)暴露通过表现出肠道、肝脏和肾脏毒性对人类和动物健康具有致癌作用。目前,ZEA诱导细胞凋亡和组织损伤的潜在机制尚不清楚。在这项研究中,我们旨在通过聚集规律间隔短回文重复(CRISPR)-Cas9筛选来鉴定调节ZEA细胞反应的基因,并进一步验证新的基因功能,以阐明体内和体外特定生物过程的分子机制。通过CRISPR激活筛选获得了2株抗ZEA细胞株Ov-KCNJ4和Ov-KCNJ12,这2株细胞株对ZEA的抗性和生长速率发生了显著变化。结果表明,ZEA可与细胞膜蛋白KCNJ4和KCNJ12相互作用,诱导细胞周期阻滞、DNA复制中断和碱基切除修复。KCNJ4和KCNJ12的过表达通过调节细胞周期来中和毒性,通过减弱KCNJ4- mitokatp通路中的氧化应激损伤来维持线粒体形态和功能,从而参与ZEA抗性。体内实验表明,AAV-KCNJ4通过改善线粒体功能,显著改善zea诱导的肾损害,提高抗氧化酶活性。我们的研究结果表明,增加钾通道水平可能是真菌毒素引起的损伤的一个假定的治疗靶点。
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来源期刊
Cell Stress
Cell Stress Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
13.50
自引率
0.00%
发文量
21
审稿时长
15 weeks
期刊介绍: Cell Stress is an open-access, peer-reviewed journal that is dedicated to publishing highly relevant research in the field of cellular pathology. The journal focuses on advancing our understanding of the molecular, mechanistic, phenotypic, and other critical aspects that underpin cellular dysfunction and disease. It specifically aims to foster cell biology research that is applicable to a range of significant human diseases, including neurodegenerative disorders, myopathies, mitochondriopathies, infectious diseases, cancer, and pathological aging. The scope of Cell Stress is broad, welcoming submissions that represent a spectrum of research from fundamental to translational and clinical studies. The journal is a valuable resource for scientists, educators, and policymakers worldwide, as well as for any individual with an interest in cellular pathology. It serves as a platform for the dissemination of research findings that are instrumental in the investigation, classification, diagnosis, and therapeutic management of major diseases. By being open-access, Cell Stress ensures that its content is freely available to a global audience, thereby promoting international scientific collaboration and accelerating the exchange of knowledge within the research community.
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