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Enhanced Expression of Anti-CD19 Chimeric Antigen Receptor in piggyBac Transposon-Engineered T Cells.


ABSTRACT: Adoptive T cell therapy using chimeric antigen receptor (CAR)-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a piggyBac transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon vector and combining irradiated activated T cells (ATCs) as feeder cells and virus-specific T cell receptor (TCR) stimulation, we achieved 51.4% ± 14% CAR+ T cells and 2.8-fold expansion after 14 culture days. Expanded CD19.CAR-T cells maintained a significant fraction of CD45RA+CCR7+ T cells and demonstrated potent antitumor activity against CD19+ leukemic cells both in vitro and in vivo. Therefore, piggyBac-based gene transfer may provide an alternative to viral gene transfer for CAR-T cell therapy.

SUBMITTER: Morita D 

PROVIDER: S-EPMC5907825 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Enhanced Expression of Anti-CD19 Chimeric Antigen Receptor in <i>piggyBac</i> Transposon-Engineered T Cells.

Morita Daisuke D   Nishio Nobuhiro N   Saito Shoji S   Tanaka Miyuki M   Kawashima Nozomu N   Okuno Yusuke Y   Suzuki Satoshi S   Matsuda Kazuyuki K   Maeda Yasuhiro Y   Wilson Matthew H MH   Dotti Gianpietro G   Rooney Cliona M CM   Takahashi Yoshiyuki Y   Nakazawa Yozo Y  

Molecular therapy. Methods & clinical development 20171222


Adoptive T cell therapy using chimeric antigen receptor (CAR)-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a <i>piggyBac</i> transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon v  ...[more]

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