Engineered Jurkat Cells for Targeting Prostate-Specific Membrane Antigen on Prostate Cancer Cells by Nanobody-Based Chimeric Antigen Receptor

Authors
Mahmoud Hassani 1
Fatemeh Hajari Taheri 2
Zahra Sharifzadeh 2
Arash Arashkia 3
Jamshid Hadjati 4
Wytske M. van Weerden 5
Shahriyar Abdoli 3
Mohammad Hossein Modarressi 1
Mohsen Abolhassani 2
1 Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
2 Department of Immunology, Hybridoma Lab, Pasteur Institute of Iran, Tehran, Iran
3 Department of Virology, Pasteur Institute of Iran, Tehran, Iran
4 Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
5 Department of Urology, Erasmus Medical Center, Rotterdam, Netherlands
10.29252/ibj.24.2.81
Abstract
Background: Recently, modification of T cells with chimeric antigen receptor (CAR) has been an attractive approach for adoptive immunotherapy of cancers. Typically, CARs contain a single-chain variable domain fragment (scFv). Most often, scfvs are derived from a monoclonal antibody of murine origin and may be a trigger for host immune system that leads to the T-cell clearance. Nanobody is a specific antigen-binding fragment derived from camelid that has great homology to human VH and low immunogenic potential. Therefore, in this study, nanobody was employed instead of scFv in CAR construct. Methods: In this study, a CAR was constructed based on a nanobody against PSMA (NBPII-CAR). At first, Jurkat cells were electroporated with NBPII-CAR, and then flow cytometry was performed for NBPII-CAR expression. For functional analysis, CAR T cells were co-cultured with prostate cancer cells and analyzed for IL-2 secretion, CD25 expression, and cell proliferation. Results: Flow cytometry results confirmed the expression of NBPII-CAR on the transfected Jurkat cells. Our data showed the specificity of engineered Jurkat cells against prostate cancer cells by not only increasing the IL-2 cytokine (about 370 pg/ml) but also expressing the T-cell activation marker CD25 (about 30%). In addition, proliferation of engineered Jurkat cells increased nearly 60% when co-cultured with LNCaP (PSMA+), as compared with DU145 (PSMA-). Conclusion: Here, we describe the ability of nanobody-based CAR to recognize PSMA that leads to the activation of Jurkat cells. This construct might be used as a promising candidate for clinical applications in prostate cancer therapy.
Keywords
Chimeric Antigen Receptor
Immunotherapy
prostate cancer
PSMA