Rapid Non-Viral Platform for Generation of Genetically Modified T Cells for Therapy

用于生成用于治疗的转基因 T 细胞的快速非病毒平台

• 批准号: 9930839
• 负责人: David L Lampi Hermanson
• 金额: $ 109.17万
• 依托单位: B-MOGEN BIOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-06 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9930839
• 关键词: Acute Lymphocytic Leukemia; Bacterial Infections; Biotechnology; CD19 gene; CD3 Antigens; Cancer Patient; Carrying Capacities; Cell Therapy; Cells; Childhood Acute Lymphocytic Leukemia; Clinical; Clinical Trials; Communicable Diseases; Cyclic GMP; DNA delivery; Data; Development; Effectiveness; Engineering; Ensure; Environment; FDA approved; Gene Delivery; Gene Expression; Gene Transfer; Generations; Genes; Genetic; Genetic Engineering; Hematopoietic; Human; Hyperactive behavior; Immunotherapeutic agent; Immunotherapy; In Vitro; Licensing; Ligation; Malignant Neoplasms; Mediating; Methods; Mus; Mycoses; Pharmacologic Substance; Phase; Protocols documentation; Protozoan Infections; Quality Control; Reproducibility; Safety; Sleeping Beauty; Southern Blotting; Subfamily lentivirinae; Surface Antigens; System; T cell therapy; T-Cell Receptor; T-Lymphocyte; Technology; Technology Transfer; Testing; Time; Transfection; Transgenes; Transgenic Organisms; Transposase; Viral; Viral Vector; Virus; Virus Diseases; base; cancer immunotherapy; cancer therapy; cell killing; chimeric antigen receptor; chimeric antigen receptor T cells; cost; cost effective; engineered T cells; gene therapy; immunogenicity; improved; in vivo; innovation; integration site; interest; mutant; non-viral gene delivery; novel; safety testing; scale up; transgene expression; tumor

Cancer immunotherapy, particularly genetically engineered adoptive T cell transfer, has shown great potential for the treatment of cancer patients. The use of T cells engineered to express a specific T cell receptor (TCRs) or chimeric antigen receptor (CARs) to treat cancer has generated durable cures for many cancer patients and has resulted in the first FDA approved CAR-T therapy to treat childhood acute lymphoblastic leukemia in 2017. Most gene therapies rely on viral methods to genetically modify human primary cells. However, viral delivery method is expensive, poorly reproducible and associated with several safety concerns including insertion in or near genes that may cause malignancy and generation of replication competent virus. Thus, non-viral DNA delivery methods, such as Sleeping Beauty and piggyBac, have been employed to generate CAR T cells. Although these non-viral delivery methods have the advantage of lower cost, immunogenicity, and regulatory considerations, they have been limited by their low transposition efficiency in primary human hematopoietic cells. In this application, we propose to rationally optimize a recently discovered transposon, TcBuster to deliver CARs to T cells. To this end, we further enhance our already very active hyperactive mutants of the TcBuster transposase and optimize the delivery of the transposon into cells. Following optimization of the TcBuster transposon system, we will combine these improvements with our proprietary methods to transfect T-cells efficiently and safely, and test the immunotherapeutic effectiveness of TcBuster delivered CAR into T cells. The successful completion of this project will result in the comprehensive methods to produce CAR T cells delivered by TcBuster which we will license to pharmaceutical companies to produce highly efficient CAR-T immunotherapy. More broadly, these methods could be expanded beyond immunotherapeutic cancer applications to various infectious diseases in which gene delivery by TcBuster in T cells could be advantageous.

癌症免疫疗法，特别是基因工程过继性 T 细胞移植，已显示出巨大的潜力 用于治疗癌症患者。使用经过改造的 T 细胞来表达特定的 T 细胞受体 (TCR) 或嵌合抗原受体（CAR）来治疗癌症已经为许多癌症患者带来了持久的治愈，并且 2017 年，FDA 批准了首个用于治疗儿童急性淋巴细胞白血病的 CAR-T 疗法。 大多数基因疗法依靠病毒方法对人类原代细胞进行基因改造。然而，病毒传递 该方法价格昂贵，重现性差，并且与多种安全问题相关，包括插入或 靠近可能导致恶性肿瘤和产生具有复制能力的病毒的基因。因此，非病毒DNA 睡美人和 PiggyBac 等递送方法已被用来生成 CAR T 细胞。 尽管这些非病毒递送方法具有成本较低、免疫原性和监管方面的优势。 考虑到这些因素，它们受到原代人类造血细胞转座效率低的限制。 在此应用中，我们建议合理优化最近发现的转座子 TcBuster 来传递 CAR 到 T 细胞。为此，我们进一步增强了已经非常活跃的 TcBuster 突变体 转座酶并优化转座子进入细胞的递送。 TcBuster 的以下优化 转座子系统，我们将把这些改进与我们专有的方法相结合来转染 T 细胞 高效、安全，并测试 TcBuster 将 CAR 递送至 T 细胞的免疫治疗效果。这 该项目的成功完成将带来生产 CAR T 细胞的综合方法 由 TcBuster 开发，我们将授权制药公司生产高效的 CAR-T 免疫疗法。更广泛地说，这些方法可以扩展到免疫治疗癌症之外 在各种传染病中的应用，其中 TcBuster 在 T 细胞中的基因传递可能是有利的。