Evaluation of the effect of the co-culture with dendritic cells in the phenotype and function of CAR-T cells derived from iPSC

• Grant number: 22/11928-3
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2023
• End date: June 30, 2025
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Jose Alexandre Marzagão Barbuto
• Grantee: Luana Correia Croda Catharino
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 24/00409-0 - Investigation of the effect promoted by MHC class II expressing cells on the differentiation of iPSC to T CD4+ lymphocytes, BE.EP.MS

Abstract

Among the immunotherapies, the CAR-T cell therapy has been standing out in the past few years, given the promising results obtained for hematologic malignancies. The insertion of a CAR (Chimeric Antigen Receptor) molecule in the T cells provides an antigen specific and MHC (Major Histocompatibility Complex) independent antigen recognition. The CAR-T cell products currently approved for clinical use are obtained from autologous sources. However, using allogeneic sources can overcome many manufacturing difficulties, such as the high cost and extensive production time, and the presence of dysfunctions and low numbers of T cells available. The use of iPSCs (Induced Pluripotent Stem Cells) for the generation of the CAR-T cells is an interesting alternative, seeing that they can be obtained by the reprogramming of different somatic cells and also present an indefinite proliferation potential and good susceptibility to genetic manipulation. For the iPSCs differentiation to T cells, various methodologies have been described in the literature. However, limitations such as the need to use murine feeder-cells and the almost exclusive generation of TCD8+ cells are yet to be addressed. Therefore, this project has the goal to obtain CAR-T cells from iPSCs using a modified methodology of co-culture with dendritic cells, aiming for a better simulation of the in vivo thymic antigen presentation, and therefore the generation of both TCD4+ cells and TCD8+ cells. Two methodologies currently available will be used as starting points: the formation of artificial thymic organoids and the feeder-free cell cultures. The CAR-T cells that present more desirable characteristics will afterwards be evaluated on preclinical studies, and the induction of further genetic modification will be assessed.