Advancing In Vitro Brain tumor Models: Harnessing Organ-on-a-Chip and Organoid Technologies for Cancer Immunotherapy Testing

• Grant number: 24/08593-5
• Support Opportunities: Regular Research Grants
• Start date: January 01, 2025
• End date: December 31, 2027
• Field of knowledge: Biological Sciences - Biology
• Principal Investigator: Oswaldo Keith Okamoto
• Grantee: Oswaldo Keith Okamoto
• Principal researcher abroad: Adrian Ranga
• Institution abroad: University of Leuven, Leuven (KU Leuven), Belgium
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Amanda Faria Assoni ; Ma Hui Ling ; Mayana Zatz ; Raiane de Oliveira Ferreira ; Vitória Alves de Lima

Abstract

Glioblastoma multiforme (GBM) remains a formidable challenge in oncology, with limited treatment options and poor prognosis. In recent years, the emergence of immunotherapies has presented new treatment strategies based on immunotherapies, with promising results for more specific treatments that provide less cytotoxicity for patients. In particular, the development of Chimeric Antigen Receptors (CARs) for expression in immune cells represent a new approach in cancer treatment, enabling the engineering of certain cell types, like T lymphocytes and Natural Killer cells, to target specific tumor antigens. Natural killer (NK) cells are innate immune cells which are human leukocyte antigen (HLA)-unrestricted and able to kill infected or cancerous cells without the need of antigen priming, and have shown promise in cancer immunotherapy once they are safe to be used as "off-the-shelf" therapy as no graft-versus-host disease (GvHD) cases have been reported so far, as well as presenting lower risk of causing cytokine release syndrome (CRS) and neurotoxicity. Recent studies have also shown promise in utilizing oncolytic viruses to enhance the efficacy of immunotherapy by promoting intratumoral immune cell infiltration and activation, thereby sensitizing tumors to immune checkpoint blockade, however it is completely unknown whether the results are translated in the human glioblastoma context. Due to the limitations of animal models in recapitulating human physiology, there is an increasing pursuit for alternative in vitro preclinical models to better predict treatment efficacy and safety.Building upon these findings, we propose to build a biomimetic on-chip glioblastoma model and investigate the potential of Zika virus (ZIKV) to enhance CAR-engineered NK cell therapy for GBM.We hypothesize that ZIKV-induced oncolytic effects may synergize with CAR-NK cells to enhance tumor cell killing and improve outcomes in GBM, while also assessing possible on-target, off-tumor toxicity Leveraging our proposed advanced GBM organoid model enriched with human induced pluripotent stem cell (hiPSC)-derived brain organoids, GBM cells, and microglia, combined with a vascularized microfluidic system, we aim to investigate the interplay between CAR-NK cells, ZIKV, and the GBM microenvironment. Our dynamic 3D model provides a unique platform to evaluate the safety and efficacy of CAR-NK therapy in the context of ZIKV infection, mimicking key aspects of tumor and brain parenchyma interactions observed in vivo. By assessing intratumoral CAR-NK cell infiltration, the impact of CAR-NK cells on tumor growth and spreading into brain organoids, and toxicity towards non-tumoral cells, we aim to determine whether combination of ZIKV-induced oncolysis with CAR-NK cell activity enhances selective GBM cell killing.This comprehensive approach will not only advance our understanding of the complex interactions between ZIKV, CAR-NK cells, and the GBM microenvironment but also inform the development of innovative immunotherapeutic strategies for the treatment of GBM. Ultimately, our findings may have significant implications for the clinical translation of ZIKV and CAR-NK therapy in GBM and other solid tumors. (AU)