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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要尽管嵌合抗原受体 (CAR) T 细胞疗法在临床上取得了前所未有的成功，但其广泛应用应用受到冗长且劳动密集型的离体制造程序的限制，这些程序导致：（i）成本高； (ii) 延迟向疾病快速进展的患者输注 CAR 细胞；以及 (iii) 具有异质性的 CAR 细胞；组成和终末分化，这限制了它们的植入和持久性，有明确的科学依据。以及对改善 CAR T 细胞生产方法的医疗需求，包括减少细胞加工的方法时间，降低制造成本，并减少 CAR 细胞分化最近，我们的实验室合作。开发了一种名为 MASTER（用于 T 细胞的多功能藻酸盐支架）的 CAR 细胞生产新技术 MASTER 由与 αCD3 结合的干燥大孔藻酸盐材料组成和 αCD28 抗体，并使用 MASTER 技术封装白细胞介素信号传导。涉及将新鲜分离的、未激活的患者 PBMC 与编码 CAR 的逆转录病毒载体一起接种植入支架后，MASTER 会介导 CAR 的每一步。生产过程，从而消除了当前αCD3/αCD28预激活的标准程序步骤，通过旋转接种和白细胞介素介导的 CAR 扩增进行病毒转导体外 MASTER 生成的 CAR。与金标准生成的 CAR 细胞相比，细胞表现出细胞分化减少，体内 MASTER 生成的 CAR 细胞表现出远远优于体内细胞的效果。持久性、增强的抗肿瘤功效以及在再次攻击后更有效地预防肿瘤生长。该系统的效用有两个：1）作为一种变革性的治疗技术，创造了增强且负担得起的治疗效果用于癌症治疗的 CAR 疗法，2) 作为一种研究工具，可实现快速开发、原型设计和测试我们组建了一支由专家组成的专注的多学科团队。生物材料和药物输送（Brudno），病毒工程和蛋白质生产专家（Birnbaum），两名临床 CAR 细胞生产专家 (Chen, Roy) 和专注于 CAR 细胞疗法的临床医生 (Grover In)。该提案我们寻求进一步开发和验证 MASTER 支架以及相关方法它们已准备好供研究和临床社区（包括相关领域的研究人员）广泛使用渴望在 CAR 领域开展工作的领域，但由于体内测试 CAR 构建的障碍而望而却步。变革性的初步数据表明，货架稳定的 MASTER 支架优于传统 CAR 淋巴瘤、原位胰腺癌以及转移性肺和卵巢癌临床前小鼠模型中的细胞该提案将在不同规模的广泛供体中验证 MASTER 支架，多个病毒载体和 CAR 构建体并描述 MASTER 产生的表型和功能这些目标的成功实现将推动我们实现低成本和低成本的最终愿景。针对液体和实体瘤的可调节 CAR 细胞生成，并可能超越肿瘤学领域。