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Review
. 2022 Jun 9:13:867013.
doi: 10.3389/fimmu.2022.867013. eCollection 2022.

The Past, Present, and Future of Non-Viral CAR T Cells

Alex Moretti  1 , Marianna Ponzo  1 , Charles A Nicolette  2 , Irina Y Tcherepanova  2 , Andrea Biondi  1   3   4 , Chiara F Magnani  1   5
Affiliations
Review

The Past, Present, and Future of Non-Viral CAR T Cells

Alex Moretti et al. Front Immunol. .

Abstract

Adoptive transfer of chimeric antigen receptor (CAR) T lymphocytes is a powerful technology that has revolutionized the way we conceive immunotherapy. The impressive clinical results of complete and prolonged response in refractory and relapsed diseases have shifted the landscape of treatment for hematological malignancies, particularly those of lymphoid origin, and opens up new possibilities for the treatment of solid neoplasms. However, the widening use of cell therapy is hampered by the accessibility to viral vectors that are commonly used for T cell transfection. In the era of messenger RNA (mRNA) vaccines and CRISPR/Cas (clustered regularly interspaced short palindromic repeat-CRISPR-associated) precise genome editing, novel and virus-free methods for T cell engineering are emerging as a more versatile, flexible, and sustainable alternative for next-generation CAR T cell manufacturing. Here, we discuss how the use of non-viral vectors can address some of the limitations of the viral methods of gene transfer and allow us to deliver genetic information in a stable, effective and straightforward manner. In particular, we address the main transposon systems such as Sleeping Beauty (SB) and piggyBac (PB), the utilization of mRNA, and innovative approaches of nanotechnology like Lipid-based and Polymer-based DNA nanocarriers and nanovectors. We also describe the most relevant preclinical data that have recently led to the use of non-viral gene therapy in emerging clinical trials, and the related safety and efficacy aspects. We will also provide practical considerations for future trials to enable successful and safe cell therapy with non-viral methods for CAR T cell generation.

Keywords: adoptive cell transfer; cancer therapy; chimeric antigen receptor (CAR T); gene therapy; immunotherapy; mRNA; non-viral vectors; transposons.

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Conflict of interest statement

Authors CM and AB are inventors on a patent related to non-viral CAR-T cell therapy (European patent application 15801344), filed by the Tettamanti Foundation, and licensed to CoImmune. Authors CN and IT are executive officers of CoImmune. Author AB has received fundings from AIRC 5x1000 2018-21147, PRIN 2017-NAZ-0412, and the Ministero della Salute CAR-T project of the “Rete Oncologica”. The remaining authors declare that the research was conducted in the absence of any commercial or financial conflict of interests.

Figures

Figure 1
Figure 1
Worldwide approval status of CAR T cell drugs.
Figure 2
Figure 2
SB and PB mediated integration.
Figure 3
Figure 3
mRNA vector.
Figure 4
Figure 4
Non-viral CAR T cell generation.
See this image and copyright information in PMC

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