Characterizing piggyBat—a transposase for genetic modification of T cells

Gaurav Sutrave; Ning Xu; Tiffany C.Y. Tang; Alla Dolnikov; Brian Gloss; David J. Gottlieb; Kenneth P. Micklethwaite; Kavitha Gowrishankar

Molecular Therapy: Methods & Clinical Development (Jun 2022)

Characterizing piggyBat—a transposase for genetic modification of T cells

Gaurav Sutrave,
Ning Xu,
Tiffany C.Y. Tang,
Alla Dolnikov,
Brian Gloss,
David J. Gottlieb,
Kenneth P. Micklethwaite,
Kavitha Gowrishankar

Affiliations

Gaurav Sutrave: Westmead Institute for Medical Research, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia
Ning Xu: Children's Cancer Institute, University of New South Wales, Sydney, NSW, Australia; School of Women's and Children's Health, UNSW Sydney, Sydney, NSW, Australia
Tiffany C.Y. Tang: Children's Cancer Institute, University of New South Wales, Sydney, NSW, Australia; School of Women's and Children's Health, UNSW Sydney, Sydney, NSW, Australia
Alla Dolnikov: Children's Cancer Institute, University of New South Wales, Sydney, NSW, Australia; School of Women's and Children's Health, UNSW Sydney, Sydney, NSW, Australia
Brian Gloss: Westmead Institute for Medical Research, Sydney, NSW, Australia
David J. Gottlieb: Westmead Institute for Medical Research, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia; Blood Transplant and Cell Therapies Program, Department of Haematology, Westmead Hospital, Hawkesbury Road, Westmead, NSW 2145, Australia
Kenneth P. Micklethwaite: Westmead Institute for Medical Research, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia; Blood Transplant and Cell Therapies Program, Department of Haematology, Westmead Hospital, Hawkesbury Road, Westmead, NSW 2145, Australia; Blood Transplant and Cell Therapies Laboratory, NSW Health Pathology – ICPMR Westmead, Sydney, NSW, Australia; Corresponding author Kenneth Micklethwaite, Blood Transplant and Cell Therapies Program, Department of Haematology, Westmead Hospital, Hawkesbury Road, Westmead, NSW 2145, Australia.
Kavitha Gowrishankar: Westmead Institute for Medical Research, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia

Journal volume & issue: Vol. 25
pp. 250 – 263

Abstract

Read online

Chimeric antigen receptor (CAR) T cells targeting CD19 have demonstrated remarkable efficacy in the treatment of B cell malignancies. Current CAR T cell manufacturing protocols are complex and costly due to their reliance on viral vectors. Non-viral systems of genetic modification, such as with transposase and transposon systems, offer a potential streamlined alternative for CAR T cell manufacture and are currently being evaluated in clinical trials. In this study, we utilized the previously described transposase from the little brown bat, designated piggyBat, for production of CD19-specific CAR T cells. PiggyBat demonstrates efficient CAR transgene delivery, with a relatively low variability in integration copy number across a range of manufacturing conditions as well as a similar integration site profile to super-piggyBac transposon and viral vectors. PiggyBat-generated CAR T cells demonstrate CD19-specific cytotoxic efficacy in vitro and in vivo. These data demonstrate that alternative, naturally occurring DNA transposons can be efficiently re-tooled to be exploited in real-world applications.

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

About the journal

Abstract

Keywords