Combinatorial CRISPR-Cas9 Metabolic Screens Reveal Critical Redox Control Points Dependent on the KEAP1-NRF2 Regulatory Axis

被引：72

作者：

Zhao, Dongxin ^{[1
]}

Badur, Mehmet G. ^{[1
]}

Luebeck, Jens ^{[2
]}

Magana, Jose H. ^{[1
]}

Birmingham, Amanda ^{[3
]}

Sasik, Roman ^{[3
]}

Ahn, Christopher S. ^{[1
]}

Ideker, Trey ^{[4
,5
]}

Metallo, Christian M. ^{[1
,5
]}

Mali, Prashant ^{[1
]}

机构：

[1] Univ Calif San Diego, Dept Bioengn, La Jolla, CA 92093 USA

[2] Univ Calif San Diego, Bioinformat & Syst Biol Grad Program, La Jolla, CA 92093 USA

[3] Univ Calif San Diego, Ctr Computat Biol & Bioinformat, La Jolla, CA 92093 USA

[4] Univ Calif San Diego, Dept Med, Div Genet, La Jolla, CA 92093 USA

[5] Univ Calif San Diego, Moores Canc Ctr, La Jolla, CA 92093 USA

来源：

MOLECULAR CELL | 2018年 / 69卷 / 04期

基金：

美国国家科学基金会;

关键词：

OXIDATIVE STRESS; PYRUVATE-KINASE; CANCER; NRF2; GENES; IDENTIFICATION; PATHWAYS; DELETION; GLUCOSE; SERINE;

D O I：

10.1016/j.molcel.2018.01.017

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (PPP). 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 was validated using C-13 and H-2 isotope tracing, and these assays together revealed key nodes controlling redox homeostasis along the KEAP-NRF2 signaling axis. Specifically, targeting KEAP1 in combination with oxidative PPP genes 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.

引用

页码：699 / +

页数：16

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