MASTER Scaffolds for Rapid, Single-Step Manufacture and Prototyping of CAR-T cells

用于快速、单步制造 CAR-T 细胞和原型制作的 MASTER 支架

• 批准号: 10713795
• 负责人: Yevgeny Brudno
• 金额: $ 37.94万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-17 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713795
• 关键词: Activities of Daily Living; Alginates; Antibodies; Area; B lymphoid malignancy; Back; Biocompatible Materials; CAR T cell therapy; CD28 gene; CD3 Antigens; Cancer Patient; Cell Count; Cell Differentiation process; Cell Therapy; Cell physiology; Cells; Clinic; Clinical; Clinical Protocols; Communities; Data; Development; Disease Progression; Disseminated Malignant Neoplasm; Drug Delivery Systems; Dryness; Encapsulated; Engineering; Engraftment; Generations; Goals; Implant; In Vitro; Interleukins; Liquid substance; Lung Neoplasms; Lymphoma; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Measures; Mediating; Medical; Methods; Modeling; Names; Oncology; Pancreatic Adenocarcinoma; Patient Isolation; Patients; Performance; Peripheral Blood Mononuclear Cell; Phenotype; Prevention; Procedures; Process; Production; Proliferating; Proteins; Public Health; Receptor Cell; Research; Research Personnel; Retroviral Vector; Signal Transduction; Solid; Solid Neoplasm; Specialist; Speed; System; T-Cell Activation; T-Lymphocyte; Technology; Testing; Therapeutic; Translations; Treatment Efficacy; Viral; Viral Vector; Virus; Visual impairment; Work; barrier to testing; bioscaffold; cancer care; cancer therapy; cellular transduction; chimeric antigen receptor; chimeric antigen receptor T cells; cost; design; engineered T cells; improved; in vivo; innovation; manufacture; manufacturing cost; manufacturing process; mouse model; multidisciplinary; new technology; novel; ovarian neoplasm; pre-clinical; prevent; procedure cost; prototype; response; retroviral transduction; scaffold; success; therapeutic candidate; tool; transcriptome sequencing; transduction efficiency; tumor; tumor growth

PROJECT SUMMARY Despite the unprecedented clinical success of chimeric antigen receptor (CAR) T cell therapy, its widespread application is limited by lengthy and labor-intensive ex vivo manufacturing procedures that result in: (i) high cost; (ii) delays to infuse CAR cells to patients with rapidly progressing disease; and (iii) CAR cells with heterogeneous composition and terminal differentiation, which limit their engraftment and persistence. There is a clear scientific and medical need for approaches to improve CAR T cell production, including methods to reduce cell processing times, reduce manufacturing costs, and reduce CAR cell differentiation. Recently, our labs collaboratively developed a new technology for CAR cell production called MASTER (Multifunctional Alginate Scaffolds for T cell Engineering and Release). MASTER consists of dry, macroporous alginate materials conjugated to αCD3 and αCD28 antibodies and encapsulating interleukin signaling. CAR generation with MASTER technology involves seeding freshly isolated, non-activated patient PBMCs together with CAR-encoding retroviral vectors and implanting scaffolds back into patients. Once implanted, MASTER mediates every step of the CAR production process, thereby eliminating the current standard procedural steps of αCD3/αCD28 pre-activation, viral transduction with spinoculation and interleukin-mediated CAR expansion. In vitro MASTER-generated CAR cells demonstrate reduced cellular differentiation as compared to CAR cells generated with gold-standard, “conventional” clinical protocols. In vivo MASTER-generated CAR cells demonstrate far superior in vivo cell persistence, enhanced anti-tumor efficacy and far superior prevention of tumor growth after rechallenge. The utility of this system is two-fold: 1) as a transformative therapeutic technology creating enhanced and affordable CAR therapy for cancer care and 2) as a research tool enabling rapid development, prototyping and testing of CAR therapeutic candidates. We have assembled a focused, multidisciplinary team comprised of an expert in biomaterials and drug delivery (Brudno), an expert in viral engineering and protein production (Birnbaum), two specialists in clinical CAR cell production (Chen, Roy) and a clinician focused on CAR cell therapies (Grover). In this proposal we seek to further develop and validate MASTER scaffolds and the associated methods to make them ready for widescale utilization by the research and clinical communities, including researchers in related areas eager to work in the CAR field but deterred by the barriers to test CAR construct in vivo. Leveraging transformative preliminary data that show that the shelf-stable MASTER scaffolds outperform conventional CAR cells in preclinical mouse models of lymphoma, orthotopic pancreatic cancer, and metastatic lung and ovarian tumors this proposal will validate MASTER scaffolds with a wide range of donors and at different scales, with multiple viral vectors and CAR constructs and delineate the phenotype and function resulting from MASTER production of CAR cells. The successful completion of these aims will propel our ultimate vision of low-cost and tunable generation of CAR cells for both liquid and solid tumors and potentially beyond the oncology space.

项目摘要 尽管嵌合抗原受体（CAR）T细胞疗法的前所未有的临床成功，但其宽度 应用程序受到冗长而劳动密集型的离体制造程序的限制，这些程序导致：（i）高成本； （ii）延迟将汽车细胞注入快速发展的疾病患者； （iii）具有异质性的汽车电池 组成和终端分化，限制了它们的植入和持久性。有一个明确的科学 和医疗需求改善CAR T细胞生产的方法，包括减少细胞处理的方法 时间，降低制造成本并降低汽车电池的差异。最近，我们的实验室合作 开发了一种用于汽车电池生产的新技术称为Master（T 细胞工程和释放）。主由与αCD3结合的干燥的大型算法材料组成 和αCD28抗体和封装白介素信号传导。用总技术产生汽车 涉及播种新鲜分离的，未激活的患者PBMC以及CAR-RECODER RETOROVOLAL载体 并将脚手架植入患者。植入后，主人会调解汽车的每一步 生产过程，从而消除了αCD3/αCD28前激活的当前标准程序步骤 病毒翻译，带有脊柱和白介素介导的CAR扩展。体外大师生成的汽车 与通过金色标准产生的汽车细胞相比，细胞表现出降低的细胞分化， “常规”临床方案。体内产生的汽车细胞在体内细胞表现出较高的表现 持久性，抗肿瘤效率提高以及对肿瘤的肿瘤生长的预防效率高。 该系统的效用是两个方面：1）作为一种变革性的治疗技术，创造了增强和负担得起的 用于癌症护理的汽车治疗和2）作为一种研究工具，可快速开发，原型制作和测试 汽车热候选人。我们已经组建了一个集中的，多学科的团队，由专家组成 生物材料和药物输送（Brudno），病毒工程和蛋白质的专家（Birnbaum），两个 临床汽车细胞生产的专家（Chen，Roy）和临床专注于汽车细胞疗法（Grover）。 我们寻求进一步开发和验证大师脚手架的这一建议以及制作的相关方法 他们为研究和临床社区的广泛利用做好了准备，包括相关研究人员 渴望在汽车场工作的地区，但由在体内测试汽车构造的障碍确定。利用 变革性的初步数据表明，货架稳定的主脚手架的表现优于常规汽车 淋巴瘤临床前小鼠模型，原位胰腺癌以及转移性肺和卵巢的细胞 肿瘤该建议将验证具有广泛捐助者的主脚手架，并且在不同的尺度上， 多个病毒矢量和汽车构造，并描述由主产生的表型和功能 汽车电池的产生。这些目标的成功完成将推动我们对低成本和 液体和实体瘤的汽车细胞可调生成，并可能超出肿瘤学空间。