ETD

• car-t
• monitoring
• outcome

Chimeric Antigen Receptor T-cell (CAR-T) therapy has emerged as an advanced treatment for patients with relapsed or refractory hematologic malignancies, achieving high response rates in diseases with otherwise limited therapeutic options, such as aggressive B-cell lymphomas and acute lymphoblastic leukemia. Despite its remarkable efficacy, CAR-T therapy is characterized by a complex toxicity profile that may significantly influence clinical outcomes and requires careful monitoring and management. Among treatment-related adverse events, cytokine release syndrome (CRS), immune effector cell–associated neurotoxicity syndrome (ICANS) and immune effector cell–associated haematotoxicity (IC-HAT) represent the most clinically relevant complications. These toxicities arise from excessive immune activation and dysregulated inflammatory responses and show substantial interpatient variability in terms of incidence, severity and timing. Their occurrence may impact not only treatment-related morbidity and mortality but also the durability of response and overall clinical benefit. In parallel with toxicity assessment, the evaluation of clinical outcomes remains central to determining the real-world effectiveness of CAR-T therapy. Increasing evidence suggests that these outcomes are closely linked to CAR-T cell expansion, persistence and functional activity in vivo. This thesis focuses on the integration of clinical outcomes and molecular monitoring in patients treated with CAR-T therapy. A droplet digital PCR (ddPCR)–based assay was developed to quantitatively monitor CAR-T cell kinetics in peripheral blood. In addition, the expression of two inflammation-related genes, Rho-Associated Coiled-Coil Containing Protein Kinase 2 (ROCK2) and Insulin-Like Growth Factor Binding Protein 6 (IGFBP6), was analyzed before and after CAR-T infusion. These genes, previously associated with immune and inflammatory pathways, were explored as potential molecular correlates of treatment-related toxicities and clinical response. By combining molecular data, clinical outcomes and toxicity profiles, this work aims to provide a more comprehensive understanding of CAR-T therapy dynamics, improve risk stratification and contribute to the development of more personalized and outcome-oriented management strategies for patients undergoing CAR-T cell treatment.