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Published on: May 12, 2017
A functional CRISPR/Cas9 screen identifies kinases that modulate FGFR inhibitor response in gastric cancer
Jiamin Chen1, John Bell2, Billy T Lau2
1Division of Oncology, Department of Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Abstract:
Some gastric cancers have FGFR2 amplifications, making them sensitive to FGFR inhibitors. However, cancer cells inevitably develop resistance despite initial response. The underlying resistance mechanism to FGFR inhibition is unclear. In this study, we applied a kinome-wide CRISPR/Cas9 screen to systematically identify kinases that are determinants of sensitivity to a potent FGFR inhibitor AZD4547 in KatoIII cells, a gastric cancer cell line with FGFR2 amplification. In total, we identified 20 kinases, involved in ILK, SRC, and EGFR signaling pathways, as determinants that alter cell sensitivity to FGFR inhibition. We functionally validated the top negatively selected and positively selected kinases, ILK and CSK, from the CRISPR/Cas9 screen using RNA interference. We observed synergistic effects on KatoIII cells as well as three additional gastric cancer cell lines with FGFR2 amplification when AZD4547 was combined with small molecular inhibitors Cpd22 and lapatinib targeting ILK and EGFR/HER2, respectively. Furthermore, we demonstrated that GSK3b is one of the downstream effectors of ILK upon FGFR inhibition. In summary, our study systematically evaluated the kinases and associated signaling pathways modulating cell response to FGFR inhibition, and for the first time, demonstrated that targeting ILK would enhance the effectiveness of AZD4547 treatment of gastric tumors with amplifications of FGFR2.
Insights
Gastric cancer cells resistant to FGFR inhibitors can be resensitized by targeting ILK. Combining FGFR inhibitor AZD4547 with an ILK inhibitor synergistically reduced cancer cell viability in FGFR2-amplified gastric cancers.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Genomics
Background:
- Gastric cancers with FGFR2 amplifications initially respond to FGFR inhibitors but develop resistance.
- The mechanisms of resistance to FGFR inhibition in gastric cancer remain largely unknown.
- Identifying novel therapeutic targets is crucial for overcoming treatment resistance.
Purpose of the Study:
- To systematically identify kinases that determine sensitivity to FGFR inhibition in gastric cancer.
- To elucidate resistance mechanisms and discover strategies to enhance FGFR inhibitor efficacy.
- To evaluate the therapeutic potential of targeting identified kinases in combination with FGFR inhibitors.
Main Methods:
- A kinome-wide CRISPR/Cas9 screen was performed in KatoIII gastric cancer cells (FGFR2 amplified) using the FGFR inhibitor AZD4547.
- Functional validation of key kinases (ILK, CSK) was conducted using RNA interference.
- Synergistic effects of drug combinations (AZD4547 with ILK and EGFR/HER2 inhibitors) were assessed in multiple gastric cancer cell lines.
Main Results:
- A CRISPR/Cas9 screen identified 20 kinases, including those in ILK, SRC, and EGFR pathways, that modulate sensitivity to FGFR inhibition.
- ILK and CSK were validated as key determinants of sensitivity to FGFR inhibition.
- Combination therapy of AZD4547 with ILK inhibitor (Cpd22) or EGFR/HER2 inhibitor (lapatinib) demonstrated synergistic effects.
- GSK3b was identified as a downstream effector of ILK signaling following FGFR inhibition.
Conclusions:
- Targeting ILK, in combination with FGFR inhibitors like AZD4547, represents a promising strategy to overcome resistance in FGFR2-amplified gastric cancers.
- This study provides novel insights into the kinase-dependent mechanisms of resistance to FGFR inhibition.
- The findings support the development of combination therapies involving ILK inhibition for gastric cancer treatment.
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