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Title
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Profiling targeted therapy responses in ROS1+ non-small cell lung cancer using genomically engineered patient-derived cell lines
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Author
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Abstract
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| Lung cancer is a heterogeneous disease that requires tailored therapeutic approaches. Significant advances in molecular understanding have led to the development of targeted therapies, such as tyrosine kinase inhibitors (TKIs) for epidermal growth factor receptor (EGFR) or anaplastic lymphoma kinase (ALK)-mutated non-small cell lung cancer (NSCLC). However, resistance to these therapies often develops, complicating treatment. This thesis focuses on addressing resistance in ROS1-rearranged NSCLC, a rare molecular subtype. ROS1-rearranged tumors, though only 2% of NSCLC cases, present unique challenges due to limited experimental models and the difficulty of studying rare mutations. The research aimed to create novel patient-derived cell lines carrying ROS1 mutations associated with TKI resistance, allowing for high-throughput drug screening. The CRISPR/Cas9 technique was used to introduce three resistance-conferring mutations (G2032R, L2026M and S1986Y) into the HCC-78 cell line expressing SLC34A2-ROS1. Drug assays using the 5 clinically approved ROS1 TKIs showed varied responses, with G2032R mutation leading to the most aggressive phenotype and highest resistance, while S1986Y showed no resistance. The mutations were then introduced into two additional patient-derived cell lines: CUTO-28 (TPM3-ROS1) and CUTO-37 (CD74-ROS1), revealing that the effect of mutations is dependent on the ROS1 fusion partner. Additionally, transcriptomic analysis of ROS1+ NSCLC revealed an upregulation in pathways related to nucleotide synthesis and cell adhesion, with IL20RB identified as a potential target for treating bone metastases. The study also profiled gene expression changes in response to repotrectinib, a next-generation TKI, and identified overexpressed genes like complement 3 (C3) and epithelial-to-mesenchymal transition (EMT)-related genes that may contribute to drug resistance in G2032R mutant cases. This thesis demonstrates the feasibility of using CRISPR/Cas9 to model rare ROS1 mutations and highlights the importance of fusion partner context in drug resistance. The new panel of 9 TKI-resistant, patient-derived ROS1+ NSCLC cell lines unveiled potential new therapeutic targets for patients with TKI-refractory or bone metastatic ROS1+ NSCLC disease. |
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Language
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English
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Publication
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Antwerp
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University of Antwerp, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Department of Medical Genetics
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2024
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DOI
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10.63028/10067/2105480151162165141
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Volume/pages
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308 p.
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Note
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Vandeweyer, Geert [Supervisor]
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op de Beeck, Ken [Supervisor]
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Van Camp, Guy [Supervisor]
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Full text (open access)
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