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细胞疗法取得了前所未有的临床成功，但其广泛的应用仍然存在。 应用受到冗长和劳动密集型的体外制造程序的限制，这导致：（i）高成本; (ii)延迟将CAR细胞输注到患有快速进展疾病的患者;和（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。有一个明确的科学 以及对改善CAR T细胞产生的方法的医学需求，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 开发了一种用于CAR细胞生产的新技术，称为MASTER（用于T细胞的多功能藻酸盐支架）， 细胞工程和释放）。MASTER由与α CD 3结合的干燥大孔藻酸盐材料组成 和α CD 28抗体和包封白细胞介素信号传导。采用MASTER技术生成CAR 涉及将新鲜分离的、未活化的患者PBMC与编码CAR的逆转录病毒载体一起接种 并将支架植入病人体内。一旦被植入，MASTER会调节CAR的每一步 生产工艺，从而消除了α CD 3/α CD 28预活化的当前标准程序步骤， 用自旋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 与用金标准产生的CAR细胞相比， “常规”临床方案。体内MASTER产生的CAR细胞显示出远优于体内细胞的上级特性。 持久性、增强的抗肿瘤功效和在再激发后对肿瘤生长的远上级预防。的 该系统的效用是双重的：1）作为一种变革性的治疗技术， 用于癌症护理的CAR疗法和2）作为研究工具，能够快速开发，原型设计和测试 CAR治疗候选人。我们组建了一个专注的多学科团队，由一名专家组成， 生物材料和药物输送（Brudno），病毒工程和蛋白质生产专家（Birnbaum）， 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）。在 我们寻求进一步开发和验证MASTER支架和相关方法， 他们准备由研究和临床社区，包括相关领域的研究人员， 渴望在CAR领域工作的领域，但由于体内测试CAR构建体的障碍而受阻。利用 初步数据表明，储存稳定的MASTER支架优于传统的CAR 淋巴瘤、原位胰腺癌和转移性肺和卵巢癌的临床前小鼠模型中的细胞 该提案将验证MASTER支架与广泛的供体和不同规模， 多个病毒载体和CAR构建体，并描述MASTER产生的表型和功能 生产CAR细胞。这些目标的成功实现将推动我们实现低成本、 CAR细胞的可调生成，用于液体和实体肿瘤，并可能超越肿瘤学领域。