Establishment of a bioprocess for functional CAR-T cell expansion: new frontier for the cancer treatment

• Grant number: 16/19741-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: December 01, 2016
• End date: July 31, 2020
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Dimas Tadeu Covas
• Grantee: Amanda Mizukami Martins
• Host Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 17/15840-5 - Strategies to enhance the potential and function of CAR-T cells for therapeutic applications, BE.EP.PD

Abstract

Redirect the immune response by genetic modification of T-cells with chimeric antigen receptor (CAR) is an emerging immunotherapeutic approach for a variety of neoplastic diseases, including lymphomas and leukemias, reaching an impressive momentum with several ongoing clinical trials. Initial data indicate the need for a large number of cells (10-100 billion) to infuse into the patient being necessary the in vitro expansion in a suitable culture system. Thus, in order for translation to clinical practice is mandatory to develop a bioprocess for cell expansion under defined, reproducible, cost-effective culture conditions ensuring proper identity, strength, purity and safety. In this context, the main objective of this work is to develop and establish a bioprocess for large-scale production and expansion of T cells modified with CAR anti-CD19 (2nd generation) evaluating different expansion culture systems for future application in the treatment of leukemias. Therefore, T cells used in this study will be previously modified with the CAR gene. These cells will be cultured in various small scale culture systems such as Erlenmeyers, spinner flasks and G-Rex flasks. By obtaining considerable cell densities, will be held the scale-up to stirred tank bioreactors, bioreactor-rocking (Wave) and hollow-fiber. This process is intended to compare the culture systems considering escalonability, phenotypic and functional stability of cells, compatibility with GMP standards, and economic aspects. Analysis of cell viability, metabolic assessment, phenotypic and functional characterization, production of intracellular cytokines and T lymphocytes cytotoxicity assay, will be performed with the expanded cells in the different systems. Furthermore, the safety of the cell during the expansion process will be evaluated by performing tests for bacteria, fungi, mycoplasma and endotoxin detection. We expected the development of an efficient, fast and reproducible platform for production and expansion of CAR-T cells that will contribute significantly to research in cell therapy. Once the platform has established, it will be validated in other specific subpopulations of lymphocytes, such NK cells. (AU)