The molecular mechanisms that instigate a healthy cell to become malignant are fueled by (epi)genetic alterations in so-called driver genes. While the Holy Grail of precision medicine is to... Show moreThe molecular mechanisms that instigate a healthy cell to become malignant are fueled by (epi)genetic alterations in so-called driver genes. While the Holy Grail of precision medicine is to identify these genetic dependencies and to target them with specific compounds in a personalized fashion, this has proven a daunting task, as tumors are exquisitely characterized by genetic instability and a mutator phenotype. Genetically engineered mouse models (GEMMs) are uniquely suited for functional in vivo validation of genotype-phenotype relationships, as they enable in vivo assessment of de novo tumorigenesis in a mammalian organism with intact immune and stromal compartments upon perturbation of (combinations of) oncogenes and/or tumor suppressor genes. Somatic modeling of cancer using CRISPR technology in vivo proved to be a true game-changing tool, allowing for rapid functional validation of candidate cancer genes enrolling from forward genetic screens and catalogs of alterations in human tumors. In this work, I showed how CRISPR approaches were deployed to precisely engineer tumorigenic events in the mouse mammary gland for dissecting oncogenic cascades, unraveling new therapeutic vulnerabilities and mechanisms of therapy resistance in different breast cancer subtypes. Show less
In this thesis I have firstly applied gene transfer technologies to the redirection of T cell specificity, by trying to overcome limitations related to non-viral gene transfer systems. In the... Show moreIn this thesis I have firstly applied gene transfer technologies to the redirection of T cell specificity, by trying to overcome limitations related to non-viral gene transfer systems. In the second part of my PhD work, I focused on genetic screens, which, I applied to understanding molecular mechanisms of escape from T cell attack and to reveal mechanisms of PD-L1 regulation. The work presented in this thesis may on the one hand facilitate the clinical application of non-viral-based gene transfer systems in T cells. On the other hand, the more fundamental discoveries related to IFN-γ-mediated tumor cell killing and PD-L1 regulation may help to further understand resistance toward immunotherapies and how to overcome them. Show less
