Combinatorial CRISPR-Cas9 Metabolic Screens Reveal Critical Redox Control Points Dependent on the KEAP1-NRF2 Regulatory Axis.
Combinatorial CRISPR-Cas9 Metabolic Screens Reveal Critical Redox Control Points Dependent on the KEAP1-NRF2 Regulatory Axis.
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DOI：
10.1016/j.molcel.2018.01.017
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发表时间：
2018-02-15
期刊：
影响因子：
16
通讯作者：
Mali P
中科院分区：
文献类型：
--
作者：
Zhao D;Badur MG;Luebeck J;Magaña JH;Birmingham A;Sasik R;Ahn CS;Ideker T;Metallo CM;Mali P
促进肿瘤生长的代谢途径已经被很好地表征，但转化事件对网络拓扑和酶的必要性的具体影响仍然知之甚少。为此，我们对代表糖酵解和戊糖磷酸途径的51个碳水化合物代谢基因进行了组合CRISPR-Cas9筛选。这种高通量方法使代谢基因的可缺性、相互作用和跨多种细胞类型的补偿的系统级审讯成为可能。利用13C和2H同位素示踪验证了特定组合敲除的代谢影响，这些分析共同揭示了沿KEAP1-NRF2信号轴控制氧化还原稳态的关键节点。具体来说，将KEAP1与氧化PPP酶结合，可以减轻这些基因敲除对生长速度的有害影响。这些结果表明，我们的整合框架，结合遗传、转录组学和通量测量，可以改善代谢网络改变的阐明，并指导基于肿瘤遗传学的代谢脆弱性的精确靶向。Zhao等人使用组合CRISPR筛选来阐明基因在癌症代谢网络中的必要性和相互作用。对细胞类型特异性必要性的检查揭示了沿KEAP1-NRF2信号轴的氧化还原代谢的关键调控。
The metabolic pathways fueling tumor growth have been well characterized, but the specific impact of transforming events on network topology and enzyme essentiality remains poorly understood. To this end, we performed combinatorial CRISPR-Cas9 screens on a set of 51 carbohydrate metabolism genes that represent glycolysis and the pentose phosphate pathway. This high-throughput methodology enabled systems-level interrogation of metabolic gene dispensability, interactions, and compensation across multiple cell types. The metabolic impact of specific combinatorial knockouts were validated using 13C and 2H isotope tracing, and, these assays together revealed key nodes controlling redox homeostasis along the KEAP1-NRF2 signaling axis. Specifically, targeting KEAP1 in combination with oxidative PPP enzymes mitigated the deleterious effects of these knockouts on growth rates. These results demonstrate how our integrated framework, combining genetic, transcriptomic, and flux measurements, can improve elucidation of metabolic network alterations, and guide precision targeting of metabolic vulnerabilities based on tumor genetics. Zhao et al. used combinatorial CRISPR screening to elucidate gene essentiality and interactions in the cancer metabolic network. Examination of cell type-specific essentiality revealed a critical regulation of redox metabolism along KEAP1-NRF2 signaling axis.
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作者：
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作者：
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