High specificity of engineered T cells with third generation CAR (CD28-4-1BB-CD3-?) based on biotin-bound monomeric streptavidin for potential tumor immunotherapy
Por:
Gallego-Valle J, Pérez-Fernández VA, Rosales-Magallares J, Gil-Manso S, Castellá M, Gonzalez-Navarro EA, Correa-Rocha R, Juan-Otero M and Pion M
Publicada:
19 sep 2024
Ahead of Print:
19 sep 2024
Resumen:
Introduction Immunotherapy has revolutionized cancer treatment, and Chimeric Antigen Receptor T cell therapy (CAR-T) is a groundbreaking approach. Traditional second-generation CAR-T therapies have achieved remarkable success in hematological malignancies, but there is still room for improvement, particularly in developing new targeting strategies. To address this limitation, engineering T cells with multi-target universal CARs (UniCARs) based on monomeric streptavidin has emerged as a versatile approach in the field of anti-tumor immunotherapy. However, no studies have been conducted on the importance of the intracellular signaling domains of such CARs and their impact on efficiency and specificityMethod Here, we developed second-generation and third-generation UniCARs based on an extracellular domain comprising an affinity-enhanced monomeric streptavidin, in addition to CD28 and 4-1BB co-stimulatory intracellular domains. These UniCAR structures rely on a biotinylated intermediary, such as an antibody, for recognizing target antigens. In co-culture assays, we performed a functional comparison between the third-generation UniCAR construct and two second-generation UniCAR variants, each incorporating either the CD28 or 4-1BB as co-stimulatory domainResults We observed that components in culture media could inhibit the binding of biotinylated antibodies to monomeric streptavidin-CARs, potentially compromising their efficacy. Furthermore, third-generation UniCAR-T cells showed robust cytolytic activity against cancer cell lines upon exposure to specific biotinylated antibodies like anti-CD19 and anti-CD20, underscoring their capability for multi-targeting. Importantly, when assessing engineered UniCAR-T cell activation upon encountering their target cells, third-generation UniCAR-T cells exhibited significantly enhanced specificity compared to second-generation CAR-T cellsDiscussion First, optimizing culture conditions would be essential before deploying UniCAR-T cells clinically. Moreover, we propose that third-generation UniCAR-T cells are excellent candidates for preclinical research due to their high specificity and multi-target anti-tumor cytotoxicity
Filiaciones:
Gallego-Valle J:
Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañon Health Research Institute Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Pérez-Fernández VA:
Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañon Health Research Institute Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Rosales-Magallares J:
Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañon Health Research Institute Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Gil-Manso S:
Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañon Health Research Institute Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Immune-Regulation Laboratory (LIR), Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Castellá M:
Immunology Service, Centre for Biomedical Diagnosis (CDB), Hospital Clínic de Barcelona (HCB), Joint Platform for Immunotherapy of Hospital Sant Joan de Deu, Barcelona, Spain
Gonzalez-Navarro EA:
Immunology Service, Centre for Biomedical Diagnosis (CDB), Hospital Clínic de Barcelona (HCB), Joint Platform for Immunotherapy of Hospital Sant Joan de Deu, Barcelona, Spain
Correa-Rocha R:
Immune-Regulation Laboratory (LIR), Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
Juan-Otero M:
Immunology Service, Centre for Biomedical Diagnosis (CDB), Hospital Clínic de Barcelona (HCB), Joint Platform for Immunotherapy of Hospital Sant Joan de Deu, Barcelona, Spain
Pion M:
Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañon Health Research Institute Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), Hospital General Gregorio Marañon, Madrid, Spain
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