Investigating functional pathway dependencies in mutation EGFR lung cancer (360G-Wellcome-207017_Z_17_Z)

EGFR inhibitors (EGFRi) are successfully used to treat non-small cell lung cancer (NSCLC), however, 10-20% of patients with EGFR mutations initially fail to respond to first-line EGFRi treatment. The mechanisms underlying intrinsic resistance to EGFRi in NSCLC patients are unclear. This project will investigate the signalling pathways essential for the survival of EGFRi-resistant NSCLC. Targeted siRNA screens have been performed in the laboratory using unique established and patient-derived cell line models which model intrinsic EGFRi resistance. The key goals of this 8 week project will be to validate preliminary RNAi screening data using an orthogonal approach. An inducible CRISPR interference (CRISPRi) system will be developed to validate the cytotoxic response observed upon knockdown of specific genes in combination with EGFRi treatment. An advantage of CRISPRi is that it is scalable, providing flexibility to examine the global signalling alterations arising from lethal interactions. CRISPRi will facilitate large-scale molecular profiling techniques including RNA-Seq and mass spectrometry based proteomics. This will provide the groundwork to ultimately define the bypass signalling pathways driving intrinsic resistance in mutant EGFR lung cancer. With this knowledge, novel therapeutic strategies can be developed as effective salvage treatments for lung cancer patients who do not respond to EGFRi treatment.