A VHH-Based Anti-MUC1 Chimeric Antigen Receptor for Specific Retargeting of Human Primary T Cells to MUC1-Positive Cancer Cells

(Pages: 502-513)
Alireza Rajabzadeh, Ph.D, 1Fatemeh Rahbarizadeh, Ph.D, 2,*Davoud Ahmadvand, Ph.D, 3Maryam Kabir Salmani, Ph.D, 4Amir Ali Hamidieh, M.D., 1,5,*
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Biochemistry, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran
Department of Stem Cell and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
Pediatric Cell Therapy Research Centre, Tehran University of Medical Sciences, Tehran, Iran
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Biochemistry, School of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran
Department of Stem Cell and Regenerative Medicine, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran
Pediatric Cell Therapy Research Centre, Tehran University of Medical Sciences, Tehran, Iran
*Corresponding Addresses: P.O.Box: 14115111 Department of Medical Biotechnology Faculty of Medical Sciences Tarbiat Modares University Tehran Iran P.O.Box 1419733151 Paediatric Cell Therapy Research Centre Tehran University of Medical Sciences Tehran Iran Emails:rahbarif@modares.ac.ir,aahamidieh@sina.tums.ac.ir
Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Rajabzadeh Alireza, Rahbarizadeh Fatemeh, Ahmadvand Davoud, Kabir Salmani Maryam, Hamidieh Amir Ali. A VHH-Based Anti-MUC1 Chimeric Antigen Receptor for Specific Retargeting of Human Primary T Cells to MUC1-Positive Cancer Cells. Cell J. 2021; 22(4): 502-513.

Abstract

Objective

Immunotherapy with redirected T cells that express a chimeric antigen receptor (CAR) is a promising prospect in cancer treatment. Most CARs use murine-derived single-chain variable fragments (scFvs) as an antigen targeting moiety, which may lead to host immunogenic responses and engineered T cell disappearance. It seems that development of less immunogenic CARs, such as CARs composed of the camelid variable domain of heavy chain antibodies (VHHs) may likely overcome this obstacle. Here, we improved the expression of the VHH-based anti-MUC1 CAR gene construct using a third generation lentiviral vector in primary human T cells and assessed its effect on antigen specific targeting, activation and cytotoxicity of redirected human T cells.

Materials and Methods

In this experimental study, we established a second generation novel CAR (VHH-based anti- MUC1 CAR) that contained a camelid-derived anti-MUC1 VHH followed by an IgG3 hinge, a CD28 transmembrane domain and signalling endodomains of CD28 and CD3+. Next, we constructed lentiviral vectors that contained this CAR gene construct using an optimized transiently virus production method and transduced it into human T cells. Cell surface expression of CAR, cytokine secretion and cytotoxic activity were assessed in the transduced CD3+T cells.

Results

The transduced T cells had high levels of surface expression of CAR. T cells that expressed anti-MUC1 CAR showed significantly increased secretion of Th1 cytokines, including IL-2, TNF alpha and IFN-γ, as well as cytotoxic activity upon recognition of MUC1 on tumour cells after co-incubation with T47D or MCF-7 (MUC1-positive) compared with A431 (MUC1-negative) or untransduced T cells.

Conclusion

Our results suggested that, given the unique properties of VHHs to prevent immunogenic responses and tonic signalling, our novel VHH-based anti-MUC1 CAR might be effective for clinical purposes in cancer immunotherapy.