Project: HuMAD (Acronym)

Full Title: Humanized Mouse for Antibody Discovery

Coordinator: Takis

CO-FINANCING POR FESR 2014-2020, Lazio Region

PROJECT DESCRIPTION

The HuMAD project aims to develop a humanized murine model for the generation of fully human monoclonal antibodies. Over the years, monoclonal antibodies (mAbs) have demonstrated their effectiveness for clinical use in a wide range of conditions, including viral and bacterial infections, autoimmunity, inflammation, as well as in the treatment of oncological diseases. Due to their high selectivity, monoclonal antibodies represent an interesting option for selectively targeting tumor cells of interest with low toxicity. Today, several mAbs are available for clinical use, and many others are under development. The first approved antibodies of murine origin did not prove to be effective due to their immunogenicity. Humanization of these antibodies, by replacing the murine immunogenic domains with their human counterparts, significantly reduced the immune response against the antibody, thereby improving their clinical efficacy. However, generating completely human antibodies would eliminate the need for the complex process of humanizing murine antibodies, which still has its own issues.
The HuMAD project is part of the larger group of RSI projects, within the framework of the Common Project called “IMMUNO,” co-financed by the Lazio Region through Lazio Innova S.p.A. and coordinated by Takis. The project aims to achieve the following objectives:

• Develop a humanized mouse model based on the transfer of human hematopoietic stem cells into severely immunocompromised mice.
• Use the model to study the development of the human immune system.
• Use the optimized model for the generation of fully human antibodies directed against a tumor antigen of clinical interest, which can then be developed into a drug:

1. Based on the entire human immunoglobulin.
2. Use scFv fragments to generate a CAR-T.

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

PURPOSE

One of the current challenges in the industry is to accelerate the development of new drugs as much as possible and to facilitate the fast and efficient access of innovative products to patients. In recent years, antibodies have become the predominant class of new drugs thanks to the advancement of protein engineering, which has allowed the development of highly specific monoclonal antibodies with reduced adverse effects. As of December 2019, 79 therapeutic mAbs have been approved by the U.S. FDA, but there is still significant growth potential, and the therapeutic antibody market has experienced explosive growth after the approval of new drugs for the treatment of various diseases, including many types of cancer, autoimmune, metabolic, and infectious diseases.
Through the interaction between academic research and industrial innovation, the project aims to expand knowledge about the development of the immune system and to establish a platform for the development of innovative drugs.
The overall goal of the project is, therefore, the generation of a humanized murine model for studying B cell development and generating therapeutic human antibodies or highly innovative products such as bispecific antibodies and CAR-T cells after vaccination.

RESULTS

The project involves the generation and optimization of a humanized murine model using human stem cells. In collaboration with Plaisant, immunocompromised mice will be inoculated with human stem cells and treated with different human cytokines to promote the engraftment and development of human hematopoietic cells. This will characterize the effect of various cytokines on the development of blood cells, particularly B cells. Once the desired development of the human immune system is achieved, the mice will be vaccinated with a human antigen present predominantly in colon-rectal tumor cells.
In collaboration with Bambino Gesù Children’s Hospital, specific B cells for the antigen will be analyzed to determine the antibody sequence that recognizes the antigen, and based on this information, Takis will produce the fully human antibody using recombinant DNA technology.
The antibody, once produced and purified, will be characterized on samples from patients with colon-rectal cancer through the collaboration with Regina Elena National Cancer Institute.

FINANCIAL SUPPORT RECEIVED

The total cost of the project eligible for funding is €673,472.50, of which Lazio Innova has granted a maximum non-repayable grant of €471,874.88. The funding resources come from the public notice “Strategic Projects” Determination No. G04052 of 4/04/2019, application code Prot. A0320-2019-28102, approved by Determination on June 10, 2020, No. G04052, BURL No. 29 of 09/04/2020, CUP: E82F20000230002.