Optimization of an in vitro experimental model to validate the anti-tumor activity of CAR-T cells in 3D culture models of glioblastoma

• Grant number: 24/19387-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: April 01, 2025
• End date: August 31, 2026
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Oswaldo Keith Okamoto
• Grantee: Letícia Maria de Oliveira Magalhães
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults, characterized by a high mortality rate and ineffective treatment options. In this scenario, CAR-T cells are considered one of the most promising emerging treatments for GBM, due to their specificity against the tumor. However, in vitro study models that accurately simulate the complex tumor conditions in vivo, in which their efficacy can be tested, still lack adaptations since they fail to represent the tumor and its microenvironment in vitro. The use of glioblastoma organoids (GBOs) has been shown to be a solution to this problem, however, in the few practices of co-culture of lymphocytes and tumor organoids described in the literature, there is dissociation of these organoids, failing in the principle of three-dimensionality of the tumor. Tumor spheroids, another type of three-dimensional model, are also valuable for studying tumor progression, surpassing monolayer culture by allowing the simulation of zones of cell proliferation, quiescence and necrosis, effects resulting from cell-cell interactions, chemical signaling and hypoxia. However, considering three-dimensional models, there are no descriptions of methodologies to determine adequate physiological proportions of effector cells and target tumor cells in co-cultures of lymphocytes with GBOs or GBM spheroids. Therefore, the project aims to optimize an in vitro experimental model that addresses these issues, to evaluate the antitumor potential of CAR-T cells in three-dimensional cell cultures. The study aims to contribute significantly to the advancement of CAR-T cell therapies by providing a robust experimental platform for the in vitro preclinical evaluation of new treatments against GBM. (AU)