Project Funding Details

Glioblastoma (GBM) is one of the most frequent and aggressive forms of brain cancer, with a median survival of 15 months. Standard treatments for GBM comprise surgery, radiotherapy, and Temozolomide-based chemotherapy, but despite great progresses in each of them, survival rates have not been significantly improved (Wu et al., 2021). As a result, there is growing interest in developing new approaches, and among them, the most promising one is cancer immunotherapy. Research in immunotherapy against GBM is thriving, and emerging data show promising results for Chimeric Antigen Receptor - T (CAR-T) cells therapy (Brown et al., 2024; Choi et al., 2024). CAR-T cells therapy, involving genetically engineered T cells targeting cancer cells, has shown potential in pre-clinical trials primarily using mouse models. However, challenges such as inadequate human-like models, molecular inter- and intra-tumoral heterogeneity, local and systemic immunosuppression (Kirschenbaum et al., 2024) have hindered progress. In this context, patient-derived organoids (PDOs) emerged as reliable preclinical 3D models, accurately reflecting tissues' in vivo functionality, genetics, molecular characteristics, and cellular structures (Schutgens et al., 2020; Lago et al., 2023). Based on this evidence, PDOs might offer a promising platform for testing CAR-T cells therapies and identifying new GBM antigens. Given the information from the literature, I believe that implementing CAR-T cells therapy in a model of patient-derived GBM organoids will increase the chances to establish more effective therapies with the ultimate goal of bringing them to clinic. The aim of this project is to implement CAR-T cells therapy for the treatment of adult glioblastoma. I will test this immunotherapeutic approach on patient-derived glioblastoma organoids, after their in deep characterization. The project is focused on three main Objectives: 1. Glioblastoma organoids (GBOs) characterization at different timepoints, via single-cell and bulk RNA sequencing (preliminary data), searching for putative cancer-associated antigens. 2. Cancer-associated antigens validation through mass spectrometry and immunohistological approaches. 3. GBOs co-culture with properly engineered CAR-T cells, alone or in mixed populations, finding the best culture conditions and assessing their killing potential. I firmly believe in the feasibility of this proposal, since I have already started establishing and characterizing patient-derived GBM organoids. Single cell and bulk RNA sequencing analyses are confirming the reliability of the GBOs culture, while giving new powerful insights regarding targetable cancer-associated antigens (Jacobs et al., 2020). Furthermore, I began co-culturing organoids with Cytokine-Induced Killer (CIK) cells, solving the initial possible technical issues and verifying their killing potential in order to implement this technique for CAR-T cells' application. Glioblastoma, the most common primary brain tumor in adults, has poor prognosis, with less than 5% of patients surviving 5 years post diagnosis. Current therapies are not significantly improving the survival outcome, constantly highlighting the need of developing new approaches. I strongly believe that immunotherapy is the most innovative and promising strategy that must be implemented for solid tumors. Moreover, patient-derived organoids unlock the possibility to exploit it considering inter- and intra-tumoral heterogeneity in proper human-recapitulating cellular and genetical assets. This project is pivotal in initiating the against-cancer personalized medicine era.