CAR T Cells Generated Using Sleeping Beauty Transposon Vectors and Expanded with an EBV-Transformed Lymphoblastoid Cell Line Display Antitumor Activity In Vitro and In Vivo

Human Gene Therapy
Leonardo ChicaybamMartín H Bonamino

Abstract

Chimeric antigen receptor (CAR) T cell immunotherapy for the treatment of cancer is now an approved treatment for B cell malignancies. However, the use of viral vectors to provide long-term CAR expression is associated with high production costs and cumbersome quality controls, impacting the final cost of CAR T cell therapies. Nonviral integrative vectors, such as Sleeping Beauty (SB) transposons, provide an alternative to modify primary T cells. Therefore, we developed a protocol to expand SB-transfected 19BBζ CAR T cells using a lymphoblastoid cell line, and evaluated T cell phenotype as well as function along the T cell expansion. Electroporation of PBMCs with transposon plasmid decreased cell viability on day 1 but had a minor impact on the frequency of memory subpopulations when compared to mock condition. CAR+ lymphocytes showed increased proliferation compared to mock control and high cytotoxic activity towards CD19+ cells without significant differences in exhaustion markers expression. Moreover, CAR+ lymphocytes showed an increased frequency by the end of the stimulation cycle compared with day 1, suggesting that CAR expression confers a selective proliferation advantage. Immunodeficient NOD scid gamma chain knockout (...Continue Reading

References

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Citations

May 5, 2020·Oncoimmunology·Luiza de Macedo AbdoMartín Hernán Bonamino
Nov 17, 2019·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Eileen McGowanYiguang Lin
Feb 20, 2021·Nature Reviews. Genetics·Gavin I EllisJames L Riley
Mar 26, 2021·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Nasheed M Hossain, Patrick J Stiff
Apr 16, 2021·Journal of Immunology Research·Viktor LukjanovPavel Šimara
Jun 8, 2021·Acta Pharmaceutica Sinica. B·Hassan DanaThomas J Webster
Jun 15, 2021·Frontiers in Cellular and Infection Microbiology·Alison J Sinclair
Jun 20, 2021·Clinical and Experimental Immunology·Rachel S CooperAlasdair R Fraser

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Methods Mentioned

BETA
genetic modification
flow cytometry
fluorescence-activated cell sorting
density gradient centrifugation
FACS
xenograft

Software Mentioned

GraphPad Prism
FlowJo
CliniMACS Prodigy
Tree Star

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