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Updated: Jan 3, 2026
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
Published on: May 23, 2025
Genome-wide CRISPR screening reveals genetic modifiers of mutant EGFR dependence in human NSCLC
Hao Zeng1, Johnny Castillo-Cabrera1, Mika Manser2
1Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, United States.
Abstract:
EGFR-mutant NSCLCs frequently respond to EGFR tyrosine kinase inhibitors (TKIs). However, the responses are not durable, and the magnitude of tumor regression is variable, suggesting the existence of genetic modifiers of EGFR dependency. Here, we applied a genome-wide CRISPR-Cas9 screening to identify genetic determinants of EGFR TKI sensitivity and uncovered putative candidates. We show that knockout of RIC8A, essential for G-alpha protein activation, enhanced EGFR TKI-induced cell death. Mechanistically, we demonstrate that RIC8A is a positive regulator of YAP signaling, activation of which rescued the EGFR TKI sensitizing phenotype resulting from RIC8A knockout. We also show that knockout of ARIH2, or other components in the Cullin-5 E3 complex, conferred resistance to EGFR inhibition, in part by promoting nascent protein synthesis through METAP2. Together, these data uncover a spectrum of previously unidentified regulators of EGFR TKI sensitivity in EGFR-mutant human NSCLC, providing insights into the heterogeneity of EGFR TKI treatment responses.
Insights
Genetic modifiers of EGFR tyrosine kinase inhibitor (TKI) sensitivity were identified in EGFR-mutant non-small cell lung cancer (NSCLC). RIC8A knockout enhanced TKI sensitivity, while ARIH2 knockout conferred resistance.
Area of Science:
- Genetics
- Molecular Biology
- Oncology
Background:
- EGFR-mutant non-small cell lung cancer (NSCLC) often responds to EGFR tyrosine kinase inhibitors (TKIs).
- Treatment responses are variable and not durable, indicating genetic factors influence EGFR dependency.
- Identifying these genetic modifiers is crucial for understanding treatment heterogeneity.
Purpose of the Study:
- To identify novel genetic determinants of EGFR TKI sensitivity and resistance in EGFR-mutant NSCLC.
- To elucidate the mechanisms by which these genetic factors regulate TKI response.
Main Methods:
- Genome-wide CRISPR-Cas9 screening was employed to identify genes affecting EGFR TKI sensitivity.
- Functional assays were performed to validate candidate genes and investigate their mechanisms of action.
Main Results:
- Knockout of RIC8A enhanced EGFR TKI-induced cell death by downregulating YAP signaling.
- Knockout of ARIH2 or Cullin-5 E3 complex components conferred resistance to EGFR inhibition.
- ARIH2-mediated resistance involves promoting nascent protein synthesis via METAP2.
Conclusions:
- RIC8A and ARIH2 are novel regulators of EGFR TKI sensitivity in EGFR-mutant NSCLC.
- These findings provide insights into the genetic basis of variable TKI treatment responses.
- Uncovered regulators offer potential therapeutic targets for overcoming TKI resistance.
