TY - JOUR
T1 - Inducible mismatch repair streamlines forward genetic approaches to target identification of cytotoxic small molecules
AU - Nguyen, Thu P.
AU - Fang, Min
AU - Kim, Jiwoong
AU - Wang, Baiyun
AU - Lin, Elisa
AU - Khivansara, Vishal
AU - Barrows, Neha
AU - Rivera-Cancel, Giomar
AU - Goralski, Maria
AU - Cervantes, Christopher L.
AU - Xie, Shanhai
AU - Peterson, Johann M.
AU - Povedano, Juan Manuel
AU - Antczak, Monika I.
AU - Posner, Bruce A.
AU - Harvey, Colin J.B.
AU - Naughton, Brian T.
AU - McFadden, David G.
AU - Ready, Joseph M.
AU - De Brabander, Jef K.
AU - Nijhawan, Deepak
N1 - Publisher Copyright:
© 2023 Elsevier Ltd
PY - 2023/11/16
Y1 - 2023/11/16
N2 - Orphan cytotoxins are small molecules for which the mechanism of action (MoA) is either unknown or ambiguous. Unveiling the mechanism of these compounds may lead to useful tools for biological investigation and new therapeutic leads. In selected cases, the DNA mismatch repair-deficient colorectal cancer cell line, HCT116, has been used as a tool in forward genetic screens to identify compound-resistant mutations, which have ultimately led to target identification. To expand the utility of this approach, we engineered cancer cell lines with inducible mismatch repair deficits, thus providing temporal control over mutagenesis. By screening for compound resistance phenotypes in cells with low or high rates of mutagenesis, we increased both the specificity and sensitivity of identifying resistance mutations. Using this inducible mutagenesis system, we implicate targets for multiple orphan cytotoxins, including a natural product and compounds emerging from a high-throughput screen, thus providing a robust tool for future MoA studies.
AB - Orphan cytotoxins are small molecules for which the mechanism of action (MoA) is either unknown or ambiguous. Unveiling the mechanism of these compounds may lead to useful tools for biological investigation and new therapeutic leads. In selected cases, the DNA mismatch repair-deficient colorectal cancer cell line, HCT116, has been used as a tool in forward genetic screens to identify compound-resistant mutations, which have ultimately led to target identification. To expand the utility of this approach, we engineered cancer cell lines with inducible mismatch repair deficits, thus providing temporal control over mutagenesis. By screening for compound resistance phenotypes in cells with low or high rates of mutagenesis, we increased both the specificity and sensitivity of identifying resistance mutations. Using this inducible mutagenesis system, we implicate targets for multiple orphan cytotoxins, including a natural product and compounds emerging from a high-throughput screen, thus providing a robust tool for future MoA studies.
KW - Forward Genetics
KW - High throughput screens
KW - Mechanism of Action
KW - Molecular Glue
KW - Target Identification
KW - Targeted protein degradation
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U2 - 10.1016/j.chembiol.2023.07.017
DO - 10.1016/j.chembiol.2023.07.017
M3 - Article
C2 - 37607550
AN - SCOPUS:85176726927
SN - 2451-9456
VL - 30
SP - 1453-1467.e8
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 11
ER -