PO.CH01.02 · 化学
mtKO:用于线粒体氧化还原生物学研究的专用向导RNA文库
mtKO: A dedicated guide RNA library for mitochondria redox biology research
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作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
线粒体是多功能细胞器,对生理调节和病理进展都至关重要。在恶性癌细胞中,线粒体重编程建立了支持肿瘤生长的代谢基础,这对于癌细胞克服内在代谢异常和应激至关重要。为揭示参与癌症发展的关键线粒体通路,我们开发了线粒体基因敲除(mtKO)——一个稳健、灵活且无偏倚的CRISPR/Cas9向导RNA筛选平台,旨在系统地鉴定关键的线粒体相关功能。mtKO文库靶向参与多种线粒体引导过程的基因,包括生物合成、跨膜转运、氧化磷酸化和氧化还原调节。通过mtKO缺失(dropout)筛选,我们鉴定出线粒体抗氧化酶超氧化物歧化酶2(SOD2)对于携带异柠檬酸脱氢酶1(IDH1)致癌突变的癌细胞的适应性和存活是不可或缺的。从机制上讲,SOD2减轻了IDH1突变型胶质瘤和软骨肉瘤细胞中由功能失调的Krebs循环活动产生的线粒体活性氧(ROS)。在功能上,SOD2维持氧化还原稳态并保持线粒体完整性,从而在体外和体内控制疾病表现。总体而言,我们的研究引入了一种强大的功能基因组学方法来探究线粒体生物学,并揭示了Krebs循环缺陷型癌症中一种选择性的线粒体氧化还原脆弱性,凸显了线粒体氧化还原失衡作为潜在治疗靶点的价值。
查看英文原文 English abstract
Mitochondria are multifunctional organelles essential for both physiological regulation and pathological progression. In malignant cancer cells, mitochondrial reprogramming establishes a metabolic foundation that supports tumor growth, which is essential for cancer cells to overcome intrinsic metabolic abnormalities and stress. To uncover key mitochondrial pathways involved in cancer development, we developed mitochondrial Knockout (mtKO) - a robust, flexible, and unbiased CRISPR/Cas9 guide RNA screening platform designed to systematically identify critical mitochondria-associated functions. The mtKO library target genes involved in diverse mitochondrial-guided processes, including biosynthesis, transmembrane transport, oxidative phosphorylation, and redox regulation. Through a mtKO dropout screen, we identified the mitochondrial antioxidant enzyme superoxide dismutase 2 (SOD2) as indispensable for the fitness and survival of cancer cells harboring oncogenic mutations in isocitrate dehydrogenase 1 (IDH1). Mechanistically, SOD2 mitigates mitochondrial reactive oxygen species (ROS) generated by dysfunctional Krebs cycle activity in IDH1-mutant glioma and chondrosarcoma cells. Functionally, SOD2 maintains redox homeostasis and preserves mitochondrial integrity thereby controlling disease manifestation both in vitro and in vivo . Overall, our study introduces a powerful functional genomics approach to interrogate mitochondrial biology and uncovers a selective mitochondrial redox vulnerability in Krebs cycle-deficient cancers, highlighting mitochondrial redox imbalance as a potential therapeutic target.
利益披露 Disclosure
C. Yang, None.