TY - JOUR
T1 - Characterization of a small molecule inhibitor of disulfide reductases that induces oxidative stress and lethality in lung cancer cells
AU - Johnson, Fraser D.
AU - Ferrarone, John
AU - Liu, Alvin
AU - Brandstädter, Christina
AU - Munuganti, Ravi
AU - Farnsworth, Dylan A.
AU - Lu, Daniel
AU - Luu, Jennifer
AU - Sihota, Tianna
AU - Jansen, Sophie
AU - Nagelberg, Amy
AU - Shi, Rocky
AU - Forcina, Giovanni C.
AU - Zhang, Xu
AU - Cheng, Grace S. W.
AU - Spencer Miko, Sandra E.
AU - de Rappard-Yuswack, Georgia
AU - Sorensen, Poul H.
AU - Dixon, Scott J.
AU - Guha, Udayan
AU - Becker, Katja
AU - Djaballah, Hakim
AU - Somwar, Romel
AU - Varmus, Harold
AU - Morin, Gregg B.
AU - Lockwood, William W.
N1 - Publisher Copyright: © 2022
PY - 2022/2/8
Y1 - 2022/2/8
N2 - Phenotype-based screening can identify small molecules that elicit a desired cellular response, but additional approaches are required to characterize their targets and mechanisms of action. Here, we show that a compound termed LCS3, which selectively impairs the growth of human lung adenocarcinoma (LUAD) cells, induces oxidative stress. To identify the target that mediates this effect, we use thermal proteome profiling (TPP) and uncover the disulfide reductases GSR and TXNRD1 as targets. We confirm through enzymatic assays that LCS3 inhibits disulfide reductase activity through a reversible, uncompetitive mechanism. Further, we demonstrate that LCS3-sensitive LUAD cells are sensitive to the synergistic inhibition of glutathione and thioredoxin pathways. Lastly, a genome-wide CRISPR knockout screen identifies NQO1 loss as a mechanism of LCS3 resistance. This work highlights the ability of TPP to uncover targets of small molecules identified by high-throughput screens and demonstrates the potential therapeutic utility of inhibiting disulfide reductases in LUAD.
AB - Phenotype-based screening can identify small molecules that elicit a desired cellular response, but additional approaches are required to characterize their targets and mechanisms of action. Here, we show that a compound termed LCS3, which selectively impairs the growth of human lung adenocarcinoma (LUAD) cells, induces oxidative stress. To identify the target that mediates this effect, we use thermal proteome profiling (TPP) and uncover the disulfide reductases GSR and TXNRD1 as targets. We confirm through enzymatic assays that LCS3 inhibits disulfide reductase activity through a reversible, uncompetitive mechanism. Further, we demonstrate that LCS3-sensitive LUAD cells are sensitive to the synergistic inhibition of glutathione and thioredoxin pathways. Lastly, a genome-wide CRISPR knockout screen identifies NQO1 loss as a mechanism of LCS3 resistance. This work highlights the ability of TPP to uncover targets of small molecules identified by high-throughput screens and demonstrates the potential therapeutic utility of inhibiting disulfide reductases in LUAD.
KW - glutathione
KW - lung cancer
KW - reactive oxygen species
KW - redox homeostasis
KW - small molecule screen
KW - thermal proteome profiling
KW - thioredoxin
UR - http://www.scopus.com/inward/record.url?scp=85124220911&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.celrep.2022.110343
DO - https://doi.org/10.1016/j.celrep.2022.110343
M3 - Article
C2 - 35139387
SN - 2211-1247
VL - 38
JO - Cell reports
JF - Cell reports
IS - 6
M1 - 110343
ER -