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Minicircle-Based Engineering of Chimeric Antigen Receptor (CAR) T Cells.
- Source :
-
Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer [Recent Results Cancer Res] 2016; Vol. 209, pp. 37-50. - Publication Year :
- 2016
-
Abstract
- Plasmid DNA is being used as a pharmaceutical agent in vaccination, as well as a basic substance and starting material in gene and cell therapy, and viral vector production. Since the uncontrolled expression of backbone sequences present in such plasmids and the dissemination of antibiotic resistance genes may have profound detrimental effects, an important goal in vector development was to produce supercoiled DNA lacking bacterial backbone sequences: Minicircle (MC) DNA. The Sleeping Beauty (SB) transposon system is a non-viral gene delivery platform enabling a close-to-random profile of genomic integration. In combination, the MC platform greatly enhances SB transposition and transgene integration resulting in higher numbers of stably modified target cells. We have recently developed a strategy for MC-based SB transposition of chimeric antigen receptor (CAR) transgenes that enable improved transposition rates compared to conventional plasmids and rapid manufacturing of therapeutic CAR T cell doses (Monjezi et al. 2016). This advance enables manufacturing CAR T cells in a virus-free process that relies on SB-mediated transposition from MC DNA to accomplish gene-transfer. Advantages of this approach include a strong safety profile due to the nature of the MC itself and the genomic insertion pattern of MC-derived CAR transposons. In addition, stable transposition and high-level CAR transgene expression, as well as easy and reproducible handling, make MCs a preferred vector source for gene-transfer in advanced cellular and gene therapy. In this chapter, we will review our experience in MC-based CAR T cell engineering and discuss our recent advances in MC manufacturing to accelerate both pre-clinical and clinical implementation.
- Subjects :
- Animals
DNA Transposable Elements
Gene Expression Regulation, Neoplastic
Humans
Neoplasms genetics
Neoplasms immunology
Neoplasms metabolism
Receptors, Antigen, T-Cell biosynthesis
Receptors, Antigen, T-Cell immunology
T-Lymphocytes immunology
T-Lymphocytes metabolism
Transfection
Transgenes
Transposases genetics
Transposases metabolism
Gene Transfer Techniques
Genetic Therapy methods
Immunotherapy, Adoptive methods
Neoplasms therapy
Receptors, Antigen, T-Cell genetics
T-Lymphocytes transplantation
Subjects
Details
- Language :
- English
- ISSN :
- 0080-0015
- Volume :
- 209
- Database :
- MEDLINE
- Journal :
- Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer
- Publication Type :
- Academic Journal
- Accession number :
- 28101686
- Full Text :
- https://doi.org/10.1007/978-3-319-42934-2_3