Department of Genetics, Cell Biology, and Development, Center for Genome Engineering, Institute of Human Genetics, Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA.
Mol Ther. 2010 Apr;18(4):674-83. doi: 10.1038/mt.2010.2. Epub 2010 Jan 26.
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.
治疗性转基因可通过病毒和非病毒方法稳定地引入人体细胞。临床级重组病毒的转导具有将基因高效转移到原代细胞的潜力，并具有治疗成功的记录。然而，病毒基因治疗的广泛应用可能受到其最初高制造成本的限制，这些成本在有限数量的生产设施中产生，而且还存在非随机整合模式的倾向。转座子介导的基因转移的体外应用现在为使用病毒载体提供了一种替代方法。临床级 DNA 质粒的制备成本大大降低，免疫原性也降低，通过瞬时共表达超活性转座酶可以提高整合效率。这促进了使用 Sleeping Beauty (SB) 转座子系统设计人类试验的设计，该系统引入嵌合抗原受体 (CAR) 以重新定向人类 T 细胞的特异性。本文综述了应用 SB 系统对 T 细胞进行基因修饰以符合 I/II 期试验的现行良好生产规范 (cGMP) 的原理和安全性影响。
