Our group studies the cellular and molecular mechanisms that control the commitment and differentiation along the T-cell lineage of hematopoietic progenitors seeding the human thymus. Our goal is to obtain mechanistic information about how dysregulation of particular intrathymic physiological pathways leads to T-cell acute lymphoblastic leukemia (T-ALL), with the final aim of identifying specific molecular targets for therapeutic intervention. Focusing on the NOTCH1 pathway as a major driver of T-ALL, we recently developed a novel in vivo model of de novo generation of human T-ALL, which revealed the contribution of several NOTCH1 targets at distinct stages of T-ALL pathogenesis. In the last two years, this information was used together with proprietary monoclonal antibodies (mAbs) and patented technology to develop state-of-the-art therapeutic strategies targeting T-ALL-specific molecules, including chimeric antigen receptors- (CAR-) armed T cells, therapeutic antibody-drug conjugates (ADCs), and secreting T-cell engaging antibodies (STAbs). We also generated cutting-edge preclinical models for in vivo validation of our T-ALL immunotherapy strategies, as an obligatory step preceding clinical translation. Our results have provided promising preclinical results as proof of concept of the suitability of new targets for implementation of next-generation efficacious and safe strategies overcoming current challenges of T-ALL immunotherapy.