A non-viral gene editing platform for cell therapies and translational autoimmune disease modeling

总结 基因编辑的T淋巴细胞有望成为广泛人类的安全有效的活体疗法。 疾病，包括自身免疫性疾病。然而，目前用于工程化细胞的病毒方法， 治疗是不精确的，过于昂贵，并具有高失败率。这些弊端限制了发展 并阻止它们渗透替代市场，在这些市场中，自然的，自发的疾病 可以在IND前和IND使能研究中解决，以改善治疗结局。 该提案的重点是优化和推进一种新型的非病毒方法的开发， 高效精确地改造人类T细胞。这项创新是一种基于纳米质粒的位点特异性基因 编辑平台，使人类细胞治疗的可调制造。这个精确的基因不仅 编辑平台在细胞工程方面产生了巨大的飞跃，但由此产生的产品也将提供 持续的临床改善超过B细胞介导的自身免疫性疾病的护理标准， 目前还不存在细胞疗法。第一阶段的提案重点是优化GeneWeld的效率 位点特异性基因编辑平台，并证明使用GeneWeld重编程的体外功能 人嵌合抗原受体（CAR）-T细胞用于消除靶向B细胞群。

SUMMARY Gene edited T lymphocytes hold promise as safe and effective living therapies for a wide range of human diseases, including autoimmune disease. However, viral methods currently employed to engineer cells for therapy are imprecise, exorbitantly expensive, and have high failure rates. These drawbacks limit development of cell therapies and prevent them from penetrating alternative markets, where natural, spontaneous disease can be addressed in pre-IND and IND-enabling studies to improve therapeutic outcomes. This proposal is focused on optimizing and advancing the development of a novel, non-viral method for highly efficient and precise engineering of human T cells. The innovation is a nanoplasmid-based, site-specific gene editing platform that enables tunable manufacturing of human cell therapeutics. Not only will this precise gene editing platform yield a quantum leap forward in cellular engineering, but the resultant product will also provide sustained clinical improvements over the standard of care for B cell–mediated autoimmune diseases, for which no current cell therapies exist. This Phase I proposal is focused on optimizing the efficiency of the GeneWeld site-specific gene editing platform and demonstrating the in vitro functionality of using GeneWeld to reprogram human Chimeric Antigen Receptor (CAR)-T cells for elimination of a targeted B cell population.