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A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors
Published on: December 7, 2014
Genome-wide CRISPR Screening Identifies NFκB and c-MET as Druggable Targets to Sensitize Lenvatinib Treatment in
Ting-Chi Rebecca Wan1, Lai Wei1, Lai-Hung Cheng1
1Department of Pathology, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China; State Key Laboratory of Liver Research, The University of Hong Kong, Hong Kong, China.
Background & Aims:
Hepatocellular carcinoma (HCC), the dominant form of liver cancer, is a leading cause of cancer death worldwide. Sorafenib and lenvatinib have long been the 2 limited options of first-line treatments for patients with unresectable advanced HCC. However, the single-drug treatment strategy only shows modest survival benefit, mostly because of the survival ability of cancer cells to activate alternative pathways for compensation. In this study, we aim to identify druggable targets contributing to lenvatinib resistance and evaluate the efficacy of combining respective inhibitors and lenvatinib on HCC.
Methods:
Genome-scale clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 knockout library screening was applied on the vehicle group and lenvatinib treatment group. Identified druggable candidates were validated individually on HCC cell models. Therapeutic effects of the combined treatment of inhibitors of candidate genes and lenvatinib were evaluated in vitro and in vivo.
Results:
We successfully identified NFKB1 and MET as critical drivers for the development of lenvatinib resistance in HCC cells. By perturbing the 2 genes with either CRISPR knockout or RNA interference approaches, lenvatinib treatments were significantly sensitized. Moreover, using small molecules QNZ and cabozantinib to target NFKB1 and MET, respectively, this together with lenvatinib could synergistically induce apoptosis and suppress HCC growth in vitro and in vivo.
Conclusion:
Our results demonstrated that genome-wide CRISPR/Cas9 screening is a powerful tool for the design of rational combinational cancer therapy and provided candidate genes possible for combined treatments with lenvatinib to improve therapy efficacy.
Insights
Researchers identified NFKB1 and MET as key drivers of lenvatinib resistance in liver cancer. Combining inhibitors of these genes with lenvatinib synergistically suppressed hepatocellular carcinoma (HCC) growth, offering a promising new combination therapy strategy.
Area of Science:
- Hepatocellular carcinoma (HCC) research
- Cancer drug resistance mechanisms
- Genomic screening technologies
Background:
- Hepatocellular carcinoma (HCC) is a leading cause of cancer death, with limited first-line treatment options.
- Current treatments like lenvatinib offer modest survival benefits due to cancer cell resistance.
- Identifying targets to overcome lenvatinib resistance is crucial for improving HCC patient outcomes.
Purpose of the Study:
- To identify druggable targets that contribute to lenvatinib resistance in HCC.
- To evaluate the efficacy of combining inhibitors of these targets with lenvatinib for HCC treatment.
Main Methods:
- Genome-scale CRISPR/Cas9 knockout library screening was employed in HCC models.
- Candidate genes identified were validated using CRISPR knockout and RNA interference.
- Combination therapies using small molecule inhibitors and lenvatinib were tested in vitro and in vivo.
Main Results:
- NFKB1 and MET were identified as critical drivers of lenvatinib resistance in HCC.
- Targeting NFKB1 and MET sensitized HCC cells to lenvatinib treatment.
- Combined treatment with lenvatinib and inhibitors of NFKB1 (QNZ) and MET (cabozantinib) synergistically induced apoptosis and suppressed tumor growth.
Conclusions:
- Genome-wide CRISPR/Cas9 screening is effective for designing rational combination cancer therapies.
- NFKB1 and MET are promising targets for combination therapy with lenvatinib to enhance HCC treatment efficacy.
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