Project: GEMMA (Acronym)

Full Title: Generation of new CAR T and BiTEs to convert the Tumor Microenvironment

Coordinator: Istituti Fisioterapici Ospitalieri (Hospital Physiotherapy Institutes) – Regina Elena National Cancer Institute IRCCS

CO-FINANCING POR FESR 2014-2020, Lazio Region

PROJECT DESCRIPTION

The GEMMA project sets various ambitious goals in the field of adoptive immunotherapy of genetically modified T lymphocytes expressing chimeric antigen receptors (CAR), a therapeutic strategy against tumors that has proven effective in hematological malignancies. However, this therapeutic approach has not found application in solid tumors, mainly due to the presence of a tumor microenvironment (TME) composed of cellular components, including cancer-associated fibroblasts (CAF), extracellular matrix, and soluble factors that make it refractory and immunosuppressive. The infiltration of T lymphocytes is hindered by a physical barrier constituted by components of the extracellular matrix, of which CAFs are the major producers. Identifying CAF-related targets could influence the efficacy and duration of CAR T cell therapy, especially considering the greater genetic stability of CAFs compared to tumor cells, a characteristic that makes them less prone to the loss of antigenic expression and immune evasion.
The GEMMA project is part of a larger group of RSI projects within the Common Project called “IMMUNO,” co-financed by the Lazio Region through Lazio Innova S.p.A. and coordinated by Bambino Gesù Children’s Hospital. The project aims to achieve the following objectives:

• Identify in silico targets involved in the exclusion of T lymphocytes from the tumor.
• Validate the identified targets in a collection of tumor-associated fibroblasts isolated from samples obtained from patients with non-small-cell lung carcinoma (NSCLC).
• Construct specific antibodies against newly identified TME-related targets.
• Generate CAR T cells from single chains and perform in vitro functionality tests on 3D models.
• Develop methods for the production of bispecific antibodies (BiTEs – bi-specific T-cell engagers).
• Study the effectiveness of the treatment in an animal model of NSCLC.

The conceptual idea and the related investment program stem from the active collaboration of two Scientific Research and Treatment Institutes (Bambino Gesù Children’s Hospital and Regina Elena National Cancer Institute), a large biopharmaceutical company (Menarini Biotech), and two small companies in the biotechnology/pharmaceutical sector (Takis and Plaisant), partners in Effective Collaboration for the realization of this project.

PURPOSE

The current challenge in oncology is to pursue new paths of precision medicine, adopting innovative therapeutic strategies that combine the excellence of industrial and academic research to define treatments that take into account individual variability, molecular characteristics of the tumor and its microenvironment, and the host’s immune system.
The project aims not only to increase knowledge about the mechanisms limiting the effectiveness of CAR T cell therapy in patients with solid tumors but also to generate new drugs for innovative therapeutic strategies that modulate the tumor microenvironment.
The overall goal of the project is the identification of TME-related targets to produce reagents and drugs that can transform a “cold” TME into an environment capable of promoting the infiltration of immune cells with anti-tumor activity, particularly the penetration of administered CAR T cells.

RESULTS

IRE’s project involves the implementation of innovative computer models to identify the main characteristic factors of the TME involved in the immune exclusion process. These in silico studies will be validated in extensive cases of tumor tissues available at the IRE Biobank, with particular attention to the study of non-small-cell lung carcinoma (NSCLC), a neoplasm that remains a “big killer” despite the new successes of immunological checkpoint-blocking drugs.
We expect to identify a panel of molecules related to a TME hostile to the infiltration of T lymphocytes, which could be considered putative therapeutic targets for the industrial development of specific CAR T and BiTEs. To demonstrate the validity of the identified targets, we will create CAF models in co-culture with tumor cells in 3D, an important preclinical model for studying the TME of lung neoplasms and for evaluating the therapeutic efficacy of new molecules aimed at transforming a TME hostile to drug penetration.
In collaboration with Takis, bispecific antibodies directed against a limited panel of therapeutic targets will be developed. The generated antibodies will be particularly activating T cell antibodies (BiTE – Bispecific T-Cell Engager) and will be tested in vitro to evaluate their efficacy and specificity. Subsequently, in collaboration with OPBG, CAR T vectors directed against the selected therapeutic target will be generated. After functional in vitro studies, the efficacy of CAR T cells will be tested on a murine model of NSCLC in collaboration with Plaisant. Additionally, thanks to the collaboration with Menarini, production processes for large-scale BiTEs will be implemented.

FINANCIAL SUPPORT RECEIVED

The total cost of the project eligible for funding is €1,015,908.72, of which Lazio Innova has granted a maximum non-repayable grant of €740,690.55. The funding resources come from the public notice “Strategic Projects” Determination No. G04052 of 4/04/2019, application code Prot. A0320-2019-28097, approved by Determination on June 10, 2020, No. G06734, BURL No. 75 of 11/06/2020, CUP: E82F20000220002.