DNA Vector Laboratory, DKFZ Heidelberg, Im Neuenheimer Feld 242, Heidelberg, Germany.
Clinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, DKFZ, Heidelberg, Germany.
Sci Adv. 2021 Apr 14;7(16). doi: 10.1126/sciadv.abf1333. Print 2021 Apr.
The compelling need to provide adoptive cell therapy (ACT) to an increasing number of oncology patients within a meaningful therapeutic window makes the development of an efficient, fast, versatile, and safe genetic tool for creating recombinant T cells indispensable. In this study, we used nonintegrating minimally sized DNA vectors with an enhanced capability of generating genetically modified cells, and we demonstrate that they can be efficiently used to engineer human T lymphocytes. This vector platform contains no viral components and is capable of replicating extrachromosomally in the nucleus of dividing cells, providing persistent transgene expression in human T cells without affecting their behavior and molecular integrity. We use this technology to provide a manufacturing protocol to quickly generate chimeric antigen receptor (CAR)-T cells at clinical scale in a closed system and demonstrate their enhanced anti-tumor activity in vitro and in vivo in comparison to previously described integrating vectors.
在有意义的治疗窗口期内，为越来越多的肿瘤患者提供过继细胞疗法（ACT）的迫切需求，使得开发一种高效、快速、多功能且安全的基因工具来构建重组 T 细胞变得不可或缺。在这项研究中，我们使用了非整合的最小尺寸 DNA 载体，该载体具有增强的产生基因修饰细胞的能力，并且我们证明它们可以有效地用于工程化人类 T 淋巴细胞。该载体平台不含病毒成分，并且能够在分裂细胞的核内以附加体形式复制，在不影响其行为和分子完整性的情况下，为人类 T 细胞提供持续的转基因表达。我们使用这项技术提供了一个制造方案，以便在封闭系统中快速在临床规模上生成嵌合抗原受体（CAR）-T 细胞，并证明与之前描述的整合载体相比，它们在体外和体内具有增强的抗肿瘤活性。
