DTA

Corso Ordinario Ciclo Unico 6 Anni

ROSSARI, FEDERICO

etd-11112020-000844

Step back to go beyond: syngeneic mouse modelling of CAR T-cell therapies to overcome their limitations arisen in glioblastoma clinical trials

Cl. Sc. Sperimentali - Medicina

SCIENZE MEDICHE - SCIENZE MEDICHE

Tutor Dott.ssa ANGELONI, DEBORA
Presidente Prof. EMDIN, MICHELE
Relatore Prof. NALDINI, LUIGI
Relatore Dott.ssa COLTELLA, NADIA
Membro Dott.ssa CASIERI, VALENTINA
Membro Prof. COCEANI, FLAVIO
Membro Prof. LIONETTI, VINCENZO
Membro Prof. PASSINO, CLAUDIO
Membro Prof. RECCHIA, FABIO ANASTASIO
Membro Dott. MEOLA, MARIO
Membro Dott.ssa PETRUCCI, ILARIA
Membro Dott. GIANNONI, ALBERTO

• CAR T cells
• Chimeric antigen receptor
• GD2
• Glioblastoma multiforme
• Solid tumors
• Syngeneic models

14/12/2020;

completa

Chimeric Antigen Receptor (CAR)-engineered T cells are a cutting-edge immunotherapeutic approach for both blood and solid malignancies. While different CAR products are already or close to be clinically approved for the former, some major hurdles are limiting their function against the latter. The poor tumor trafficking and short persistence of T cells, the presence of an immunosuppressive tumor microenvironment, the occurrence of antigen loss or other forms of immune evasion and that of unpredicted toxicities represent the key limiting factors for CAR T therapies against solid cancers. Since most of those observations are directly made during clinical trials, the most widely used pre-clinical models are likely to be suboptimal. In fact, they almost entirely rely on immunocompromised mouse models with xenografted tumors and CAR T cells. Despite the clinical relevance of the targeted antigen, the CAR manufacturing process, the T cell biology and the tumor histopathology, they poorly recapitulate the fine tumor-immune system interaction, which is key to evaluate both safety and efficacy of immunotherapies, such as CAR T cells. In comparison, syngeneic mouse models, which preserve the host immune system, would be ideal to properly investigate the crosstalk among tumor cells, T cells and other immune actors; however, they commonly lack the relevance of the humanized models, therefore being poorly utilized, so far.
Given the intrinsic limitations of immunocompromised models, a multifaceted approach is here proposed to reliably investigate CAR T-cell function against glioblastoma, as candidate disease, in a syngeneic setting. First, a detailed optimization of mouse T cell culturing and lentiviral transduction is presented; second, different target antigens are identified by a “bedside to bench” approach, selecting human-mouse cross-reactive antigens that are already targeted in glioblastoma clinical trials, e.g. GD2; third, a GD2-redirected murine CAR construct is designed maintaining the target-binding region unchanged from the human counterpart; last, the function of mouse CAR T cells targeting the GD2 antigen is investigated in vivo in a new syngeneic and clinically reliable high-grade glioma model, decreasing its growth and highlighting the possibility of toxicities to occur.
Overall, the present work would serve as an alternative and improved preclinical platform to investigate new and old CAR constructs. Furthermore, this could allow for the evaluation of combination strategies in a more reliable manner, in order to gain efficacy insights to be brought to the clinic.

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