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

Hassani, Mahmoud; Hajari Taheri, Fatemeh; Sharifzadeh, Zahra; Arashkia, Arash; Hadjati, Jamshid; Van Weerden, Wytske M.; Abdoli, Shahriyar; Modarressi, Mohammad Hossein; Abolhassani, Mohsen

doi:10.29252/ibj.24.2.81

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