title: A transposon and transposase system for human application.
abstract: The stable introduction of therapeutic transgenes into human cells can be accomplished using viral and nonviral approaches. Transduction with clinical-grade recombinant viruses offers the potential of efficient gene transfer into primary cells and has a record of therapeutic successes. However, widespread application for gene therapy using viruses can be limited by their initially high cost of manufacture at a limited number of production facilities as well as a propensity for nonrandom patterns of integration. The ex vivo application of transposon-mediated gene transfer now offers an alternative to the use of viral vectors. Clinical-grade DNA plasmids can be prepared at much reduced cost and with lower immunogenicity, and the integration efficiency can be improved by the transient coexpression of a hyperactive transposase. This has facilitated the design of human trials using the Sleeping Beauty (SB) transposon system to introduce a chimeric antigen receptor (CAR) to redirect the specificity of human T cells. This review examines the rationale and safety implications of application of the SB system to genetically modify T cells to be manufactured in compliance with current good manufacturing practice (cGMP) for phase I/II trials.
authors.affiliation: Department of Genetics, Cell Biology, and Development, Center for Genome Engineering, Institute of Human Genetics, Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Perry B
authors.initials: PB
authors.lastname: Hackett
authors.forename: David A
authors.initials: DA
authors.lastname: Largaespada
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000276:immunology; D017321:Clinical Trials, Phase I as Topic; D017322:Clinical Trials, Phase II as Topic; D004251:DNA Transposable Elements / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D010957:Plasmids; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation; D014161:Transduction, Genetic / Q000379:methods*; D019895:Transposases / Q000235:genetics / Q000378:metabolism*
chemical_list: D018941:Antigens, CD19; D004251:DNA Transposable Elements; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D019895:Transposases; C494471:sleeping beauty transposase, human