Advanced development of composite gene delivery and CAR engineering systems

复合基因递送和CAR工程系统的先进开发

• 批准号: 10709085
• 负责人: Sidi Chen
• 金额: $ 41.71万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10709085
• 关键词: Acceleration; Adopted; Adoptive Cell Transfers; Advanced Development; Benchmarking; Biomedical Engineering; CTLA4 gene; Cell Survival; Cell Therapy; Cell physiology; Cells; Cellular immunotherapy; Chromosomal translocation; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Complex; Cytoplasmic Tail; DNA; DNA Transposons; Disease; Dissection; Electroporation; Elements; Engineering; Face; Functional disorder; Gene Delivery; Gene Transduction Agent; Generations; Genomics; Hematopoietic Neoplasms; Immune; Immunologic Receptors; Immunologics; Immunology; Immunotherapeutic agent; Joints; Laboratory Research; Lentivirus Vector; Ligands; Malignant Neoplasms; Measures; Memory; Messenger RNA; Metabolic; Molecular; Nature; Output; Pathway interactions; Patients; Performance; Process; Retroviral Vector; Risk; Signal Transduction; Sleeping Beauty; Solid Neoplasm; System; T-Lymphocyte; TP53 gene; Tail; Technology; Therapeutic; Toxic effect; Transposase; Tumor Antigens; Validation; adeno-associated viral vector; anticancer research; cell type; cellular engineering; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; design; exhaustion; gene delivery system; genome editing; genomic aberrations; genotoxicity; improved; in vivo; knockout gene; monomer; neoplastic cell; new technology; novel; programs; promoter; prototype; receptor; research and development; success; technology development; technology validation; therapeutic development; tool; transgene expression; tumor

SUMMARY | Cellular immunotherapy such as chimeric antigen receptor (CAR) T cells involves engineering and adoptive transfer of cells to directly target tumor cells in patient, and demonstrated clinical success. However, current cell forms of cell therapies face multiple major hurdles, centering around efficient generation of potent, specific and safe therapeutic immune cells. This R33 will address this significant problem, by developing and rigorously validating two sets of versatile gene delivery and cell engineering toolkits, to ease the therapeutic cell generation issue, and to offer a simple yet distinct approach to achieve high potency. Aim 1. Advanced development of MAJESTIC, a highly efficient composite gene delivery system. A vital part of cellular immunotherapies is therapeutic cell generation. Current approaches, including lentiviral or g-retroviral vectors, AAV, mRNA, DNA transposons, and genome editing such as CRISPR/Cas, all have their own limitations. In the first part of this R33, we will perform advanced development and validation of MAJESTIC technology (mRNA AAV-Sleeping-Beauty Joint Engineering of Stable Therapeutic Immune Cells). This system can transduce diverse immune cell types with minimal cellular toxicity, leading to highly efficient and stable therapeutic cargo delivery. Aim 2. Advanced development of synthetic fusion tails to enhance therapeutic cell function. Despite success in CAR-T therapy in hematopoietic malignancies, major challenges still exist such as tumor antigen loss, T cell exhaustion, T cell dysfunction, and poor in vivo persistence that hampered its widespread clinical potential. We seek to develop and validate a distinct approach for cellular engineering to add to the armamentarium of tools to enhance CAR immune cells against cancer. In the second part of this R33, we will advance the development of TAILFUSE technology, a novel and unique CAR engineering approach by cytoplasmic tail (CT) fusions, which reprograms CAR-T function and substantially enhanced in vivo anti-tumor efficacy. We achieved proof-of-concept development of the technologies (R21/R61 equivalent). In this project we will perform robust validation, optimization, extension and advanced development (R33). This R33 will mature these versatile tools for gene delivery and synthetic cell engineering, including quantitative performance measures, benchmarking, new capability extension, and validation of broader applications to cancer-related cell types. Success of this R33 will lead to novel technologies that will bring new capabilities, with substantial improvements over existing technologies. We anticipate wide-spread use of such technologies in laboratory research and therapeutic development settings by the field to reach transformative impact.

摘要|细胞免疫疗法，例如嵌合抗原受体（CAR）T细胞涉及工程和 细胞的产物转移以直接靶向患者的肿瘤细胞，并显示出临床成功。然而， 当前细胞疗法的细胞形式面临多个主要障碍，以有效产生有效的产生， 特异性且安全的治疗性免疫细胞。 R33将通过开发来解决这个重大问题 并严格验证两组多功能基因输送和细胞工程工具包，以简化 治疗性细胞生成问题，并提供一种简单但独特的方法来实现高效力。目的 1。高效的复合基因输送系统的高级发展。至关重要的部分 细胞免疫疗法是治疗性细胞的产生。当前的方法，包括慢病毒或G-逆转录病毒 向量，AAV，mRNA，DNA转座子和基因组编辑（例如CRISPR/CAS）都有自己的 限制。在R33的第一部分，我们将进行雄伟的高级发展和验证 技术（MRNA AAV呼吸疗程的联合工程稳定的治疗性免疫细胞）。这个系统 可以通过最小的细胞毒性转导多种免疫细胞类型，从而导致高效和稳定 治疗性货物交付。目标2。合成融合尾巴的高级开发以增强治疗性 细胞功能。尽管在造血恶性肿瘤方面的CAR-T疗法取得了成功，但仍然存在主要挑战 例如肿瘤抗原丧失，T细胞耗尽，T细胞功能障碍和体内持续性较差，这阻碍了其 广泛的临床潜力。我们寻求开发和验证一种蜂窝工程的独特方法 加入工具的武器库，以增强对癌症的CAR免疫细胞。在此R33的第二部分中， 我们将推进Tailfuse Technology的开发，这是一种新颖而独特的汽车工程方法 细胞质尾巴（CT）融合，它重新编程了CAR-T功能并在体内抗肿瘤中大大增强 功效。我们实现了技术的概念验证开发（R21/R61同等用品）。在这个项目中 我们将执行强大的验证，优化，扩展和高级开发（R33）。这个R33会成熟 这些用于基因输送和合成细胞工程的多功能工具，包括定量性能 措施，基准测试，新能力扩展以及对癌症相关细胞的更广泛应用的验证 类型。这款R33的成功将导致新型技术，这些技术将带来新的能力，并具有实质性 改进现有技术。我们预计在实验室中广泛使用此类技术 该领域的研究和治疗开发环境达到变革性影响。