Unravelling lung adenocarcinoma paralog dependencies using in vivo combinatorial CRISPR screens

• Applicant: Eiman Elwakeel, Ph.D.
• Subject Area: Hematology, Oncology ; General Genetics and Functional Genome Biology
• Term: since 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 512658514

Despite substantial advances in targeted therapeutic approaches to Lung adenocarcinoma (LuAD), the overall response of most patients with metastatic disease remains low. Genomic instability and the ensuing genetic heterogeneity is among the main factors that contributes to poor drug responses and poses an ongoing challenge to developing cancer therapeutics. Among the central oncogenic drivers of LuAD, are mutated Kirsten rat sarcoma virus (KRAS) and epidermal growth factor (EGFR). Given that the EGFR-RAS signaling pathway is comprised of many paralogous genes, perhaps it is not surprising that single gene knockouts can fail to recapitulate the effects of key therapeutic targets. Thus, identifying mutation-specific or context-specific paralog dependencies is an attractive strategy for discovering new therapeutic targets for the treatment of LuAD patients. The main goal of this proposal is to identify novel LuAD paralog gene dependencies in the setting of EGFR and KRAS mutations. I will employ a high complexity CRISPR paralog targeting library to investigate the viability effects of disrupting 4,547 genes, 8,003 paralog gene pairs. Moreover, I will develop an efficient screening system to validate such paralogous dependencies in vivo and monitor the effect of dual-gene depletion on tumor growth, metastasis and drug resistance. As the need for new therapeutic targets remains high in the setting of therapeutic resistance, I will also probe paralog gene dependencies in KRAS and EGFR-mutant LuAD cell lines that have acquired resistance to key RAS or EGFR targeting drugs (e.g., adagrasib and osimertinib). The proposed study will be the first to address LuAD paralog gene dependencies with the aim of discovering new therapeutic targets in these settings.

• DFG Programme: WBP Fellowship
• International Connection: USA
• Participating Institution: Max-Planck-Institut für Herz- und Lungenforschung; W.G. Kerkhoff-Institut
• Hosts: Professor Dr. Rajkumar Savai; Professor Dr. William Sellers