The GEMMA project has several ambitious goals in the area of adoptive immunotherapy of T lymphocytes genetically engineered to express chimeric antigen-specific receptors (CARs), an anti-tumor therapeutic strategy that has proven effective in hematologic malignancies. However, this therapeutic approach has not found its application in solid tumors mainly due to the presence of a tumor microenvironment (TME), composed of cellular components, including tumor-associated fibroblasts (CAFs), extracellular matrix, and soluble factors that make it refractory and immunosuppressive. Indeed, T lymphocyte infiltration is hindered by a physical barrier consisting of extracellular matrix components of which CAFs are the major producers. The identification of CAF-related targets could also affect the efficacy and duration of CAR T cell therapy because of the greater genetic stability of CAFs compared with cancer cells, a characteristic that makes them less prone to loss of antigenic expression and immunological evasion.

The GEMMA project is part of the larger group of CSR projects under the Joint Project called "IMMUNO," co-funded by the Lazio Region through Lazio Innova s.p.a. and coordinated by the Bambino Gesù Children's Hospital with the following objectives:

1. To identify in silico targets involved in the exclusion of T lymphocytes from the tumor
2. To validate the targets identified in a collection of tumor-associated fibroblasts isolated from samples obtained from patients with non-small cell lung cancer (NSCLC)
3. To construct specific antibodies to newly identified TME-related targets
4. To generate CAR T cells from single chain and perform in vitro functionality tests on 3D models
5. To develop methods for the production of bi-specific T-cell engagers (BiTEs) antibodies
6. To study the efficacy of treatment in the NSCLC animal model

The idea of the project and its related investment program arise from the effective collaboration of two Scientifically Oriented Research and Treatment Institutes (Ospedale Pediatrico Bambino Gesù and Istituto Nazionale Tumori Regina Elena), a large biopharmaceutical company (Menarini Biotech), and two small biotech/pharmaceutical companies (Takis and Plaisant), partners in an Effective Collaboration for realizing this project.

The current challenge in oncology is to achieve new precision medicine pathways by 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 immune system.

The project aims not only to increase knowledge about the mechanisms that limit the efficacy 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.

Thus, the overall goal of the project is to identify TME-related targets to produce reagents and drugs that can subvert a "cold" TME into a microenvironment capable of promoting the infiltration of immune system cells with antitumor activity and in particular the penetration of administered CAR T cells.

The IRE project involves the implementation of innovative computer models in order to identify the main characteristic factors of TME involved in the immune exclusion process. These in silico studies will be validated in large case series of tumor tissues available at the IRE Biobank focusing on the study of Non-small-cell lung cancer (NSCLC Non-small-cell lung carcinoma), a neoplasm that despite new successful steps of immune checkpoint blocking drugs remains a "big killer."

We plan to identify a panel of molecules related to a microenvironment hostile to T-lymphocyte infiltration that can be considered putative therapeutic targets for the industrial development of specific CAR T and BiTE. To demonstrate the validity of the identified targets, we will create models of CAFs co-cultured with tumor cells in 3D, an important preclinical model for studying the TME of lung malignancies and for evaluating the therapeutic efficacy of novel molecules aimed at subverting a TME hostile to drug penetration.

With the collaboration of Takis, bispecific antibodies directed against a narrow panel of therapeutic targets will be developed. The antibodies generated will be specifically bispecific T-cell activating antibodies (BiTE- Bispecific T-Cell Engager) and will be tested in vitro to evaluate their efficacy and specificity. Subsequently, CAR T vectors directed against the selected therapeutic target will be generated in collaboration with OPBG. After an in vitro functional study, the efficacy of CAR T cells will be tested on an NSCLC mouse model in collaboration with Plaisant. In addition, through the collaboration with Menarini, manufacturing processes for large-scale production of BiTEs will be implemented.

The total cost of the grant-eligible project is Euro 1,015,908.75, on the expenditure of which Lazio Innova has granted a maximum grant funding of Euro 740,690.55

The funding resources are from the public notice "Strategic Projects" Determination No. G04052 of 4/04/2019, application code Prot. A0320-2019-28097, approved by Determination of June 10, 2020, No. G06734, BURL No. 75 of 11/06/2020, CUP: E82F20000220002.

Oncohematology

• The Humanized Mouse for Antibody Discovery (HuMAD) project aims to develop a humanized mouse model for generating fully human monoclonal antibodies.
  HuMAD is coordinated by Takis s.r.l., in collaboration with the IRCCS Regina Elena National Cancer Institute.
  Project link
• The CARSA project aims to develop new therapies in oncohematology.
  CARSA is coordinated by the Ospedale Pediatrico Bambino Gesù in collaboration with the IRCCS Regina Elena National Cancer Institute.
  Project link
• The IMMUNO project aims to promote, manage, and disseminate the CARSA, GEMMA, HuMAD, and TraZimAb research and industrial development projects.
  IMMUNO is coordinated by the Ospedale Pediatrico Bambino Gesù in collaboration with the IRCCS Regina Elena National Cancer Institute.
  Link to the project