Journal
JOURNAL OF MEDICAL GENETICS
Volume 56, Issue 1, Pages 10-17Publisher
BMJ PUBLISHING GROUP
DOI: 10.1136/jmedgenet-2018-105405
Keywords
bacteria-free minicircle vector; integration free car-t cells; cell viability; human Cd34+Hscs; human es cells
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Funding
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDA01010409, XDA16010205]
- State 863 project [2015AA020307]
- National Natural Science Foundation of China [31471215, 81773269]
- National Key Research and Development Program of China [2016YFA0101402]
- Young Thousand Talent Project
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Background Chimeric antigen receptor T (CAR-T) cells engineered with lentiviral and retroviral vectors have been successfully applied to treat patients with B cell malignancy. However, viral integration in T cells has the potential risk of mutagenesis, and viral vector production demands effort and is costly. Using non-integrative episomal vector such as minicircle vector to generate integration-free CAR-T cells is an attractive option. Methods and results We established a novel method to generate minicircle vector within a few hours using simple molecular biology techniques. Since no bacteria is involved, we named these vectors bacteria-free (BF) minicircle. In comparison with plasmids, BF minicircle vector enabled higher transgene expression and improved cell viability in human cell line, stem cells and primary T cells. Using BF minicircle vector, we generated integration-free CAR-T cells, which eliminated cancer cells efficiently both in vitro and in vivo. Conclusion BF minicircle vector will be useful in basic research as well as in clinical applications such as CAR-T and gene therapy. Although the transgene expression of minicircle vector lasts apparently shorter than that of insertional lentivirus, multiple rounds of BF minicircle CAR-T cell infusion could eliminate cancer cells efficiently. On the other hand, a relatively shorter CAR-T cell persistence provides an opportunity to avoid serious side effects such as cytokine storm or on-target off-tumour toxicity.
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