@article{42545e07d9734b7bbf2b0a7be052993c,
title = "GCN2 adapts protein synthesis to scavenging-dependent growth",
abstract = "Pancreatic cancer cells with limited access to free amino acids can grow by scavenging extracellular protein. In a murine model of pancreatic cancer, we performed a genome-wide CRISPR screen for genes required for scavenging-dependent growth. The screen identified key mediators of macropinocytosis, peripheral lysosome positioning, endosome-lysosome fusion, lysosomal protein catabolism, and translational control. The top hit was GCN2, a kinase that suppresses translation initiation upon amino acid depletion. Using isotope tracers, we show that GCN2 is not required for protein scavenging. Instead, GCN2 prevents ribosome stalling but without slowing protein synthesis; cells still use all of the limiting amino acids as they emerge from lysosomes. GCN2 also adapts gene expression to the nutrient-poor environment, reorienting protein synthesis away from ribosomes and toward lysosomal hydrolases, such as cathepsin L. GCN2, cathepsin L, and the other genes identified in the screen are potential therapeutic targets in pancreatic cancer.",
keywords = "Cathepsin L, GCN2, PDAC, lysosomes, macropinocytosis, protein scavenging, protein synthesis, translation",
author = "Michel Nofal and Tim Wang and Lifeng Yang and Jankowski, {Connor S.R.} and {Hsin-Jung Li}, Sophia and Seunghun Han and Lance Parsons and Frese, {Alexander N.} and Zemer Gitai and Anthony, {Tracy G.} and Martin W{\"u}hr and Sabatini, {David M.} and Rabinowitz, {Joshua D.}",
note = "Funding Information: The authors thank Britt Adamson, Amanda Amodeo, Eileen White, Shawn Davidson, Bryan King, and Robert Marmion for useful discussion, Lillia Ryazanova (supported by the Lewis-Sigler Collaboration Fund and the Princeton Catalysis Initiative) for help with proteomics, and Gary Laevsky for imaging support. The authors also thank Craig Thompson and Scott Lowe for generously providing cell lines. lentiCRISPR v2 was a gift from Feng Zhang (Addgene plasmid # 52961). This work was supported by grants from the NIH (R01 CA163591 and DP1 DK113643) and from the Stand Up To Cancer (Dream Team Translational research grant SU2C-AACR-DT2016) to J.D.R. by fellowship support from the NIH (F31 CA186513) to M.N. by support from the Howard Hughes Medical Institute to D.M.S. by an NIH grant (R35GM128813) to M.W. by support from the NIH (T32GM007388) to A.N.F. by an NIH grant (R01 DK109714) to T.G.A. and by an NIH grant (DP1 AI124669) to Z.G. Stand Up To Cancer is a program of the Entertainment Industry Foundation, administered by the American Association for Cancer Research. M.N. T.W. D.M.S. and J.D.R. designed the genome-wide screens; M.N. and T.W. carried out the screens; M.N. and T.W. analyzed the CRISPR screen data. M.N. and S.H. generated knockout and re-expression cell lines. M.N. carried out the proliferation studies and analyzed the data. M.N. S.H.-J.L. Z.G. T.G.A. and J.D.R. designed the polysome profiling and ribosome profiling experiments. M.N. S.H.-J.L. and A.N.F. carried out the polysome profiling and ribosome profiling experiments. M.N. analyzed the polysome profiling data. M.N. and L.P. analyzed the ribosome profiling data. M.N. and J.D.R. designed the stable isotope tracer experiments. M.N. carried out the stable isotope tracer experiments and analyzed the data. M.N. and J.D.R. designed the radioactive tracer experiments. M.N. and C.S.R.J. carried out the radioactive tracer experiments and analyzed the data. M.N. M.W. and J.D.R. designed the proteomics experiments. M.N. carried out the proteomics experiments. M.N. and M.W. analyzed the proteomics data. M.N. and J.D.R. designed the inhibitor experiments. M.N. and L.Y. carried out the inhibitor experiments and analyzed the data. M.N. and J.D.R. wrote the paper. J.D.R. is an advisor and stockholder in Kadmon Pharmaceuticals, Colorado Research Partners, L.E.A.F. Pharmaceuticals, Bantam Pharmaceuticals, Barer Institute, and Rafael Pharmaceuticals; a paid consultant of Pfizer; a founder, director, and stockholder of Farber Partners, Serien Therapeutics, and Sofro Pharmaceuticals; a founder and stockholder in Toran Therapeutics and Raze Therapeutics; inventor of patents held by Princeton University; and a director of the Princeton University-PKU Shenzhen collaboration. D.M.S. and T.W. are co-founders of KSQ Therapeutics, which is using CRISPR-based genetic screens to identify drug targets. Z.G. is the founder of ArrePath. Funding Information: The authors thank Britt Adamson, Amanda Amodeo, Eileen White, Shawn Davidson, Bryan King, and Robert Marmion for useful discussion, Lillia Ryazanova (supported by the Lewis-Sigler Collaboration Fund and the Princeton Catalysis Initiative) for help with proteomics, and Gary Laevsky for imaging support. The authors also thank Craig Thompson and Scott Lowe for generously providing cell lines. lentiCRISPR v2 was a gift from Feng Zhang (Addgene plasmid # 52961). This work was supported by grants from the NIH ( R01 CA163591 and DP1 DK113643 ) and from the Stand Up To Cancer (Dream Team Translational research grant SU2C-AACR-DT2016 ) to J.D.R., by fellowship support from the NIH ( F31 CA186513 ) to M.N., by support from the Howard Hughes Medical Institute to D.M.S., by an NIH grant ( R35GM128813 ) to M.W., by support from the NIH (T32GM007388) to A.N.F., by an NIH grant ( R01 DK109714 ) to T.G.A., and by an NIH grant ( DP1 AI124669 ) to Z.G. Stand Up To Cancer is a program of the Entertainment Industry Foundation, administered by the American Association for Cancer Research. Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2022",
month = feb,
day = "16",
doi = "10.1016/j.cels.2021.09.014",
language = "English (US)",
volume = "13",
pages = "158--172.e9",
journal = "Cell Systems",
issn = "2405-4712",
publisher = "Cell Press",
number = "2",
}