Biomaterial Scaffolds for Ex Vivo and In Situ CAR-T Cell Production

用于离体和原位 CAR-T 细胞生产的生物材料支架

基本信息

批准号：
10591482
负责人：
Yevgeny Brudno
金额：
$ 37.63万
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：
2021
资助国家：
美国
起止时间：
2021-04-16 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10591482
关键词：
3-Dimensional Achievement Adopted Alginates Allogenic Antibodies Architecture Autologous B lymphoid malignancy Biocompatible Materials Biological Assay CAR T cell therapy CD28 gene CD3 Antigens CXCL10 gene Cell Count Cell Differentiation process Cell Reprogramming Cell Therapy Cell physiology Cells Cellular biology Centrifugation Characteristics Circulation Clinical Clinical Data Coculture Techniques Cytotoxic T-Lymphocytes Data Devices Disease Disease Progression Dose Drug Delivery Systems Encapsulated Engineering Engraftment Ensure Generations Genetic Goals Hematopoietic Neoplasms Immune system Implant In Situ In Vitro Interleukin-2 Interleukins Kinetics Lentivirus Lentivirus Vector Longevity Lymphocyte Lymphoma Mediating Medical Methods Modeling Mus Patients Peripheral Blood Mononuclear Cell Phenotype Physiological Polybrene Porosity Procedures Process Production Proliferating Proteins Protocols documentation Research Retroviral Vector Signal Transduction Solid Neoplasm Specialist Specificity System T-Cell Activation T-Lymphocyte Technology Testing Therapeutic Time Translating Tumor Burden Viral Viral Vector Xenograft Model biomaterial compatibility bioscaffold cellular transduction chimeric antigen receptor chimeric antigen receptor T cells clinically relevant confocal imaging cost crosslink cytokine density design graft vs host disease immunogenicity implantation improved in vivo manufacture manufacturing facility manufacturing process mechanical properties multidisciplinary preclinical efficacy prevent procedure cost programs recruit response scaffold success transduction efficiency translational potential tumor

项目摘要

PROJECT SUMMARY Despite unprecedented clinical success of chimeric antigen receptor (CAR)-T cell therapy against tumors, widespread application is limited by lengthy and labor-intensive ex vivo manufacturing procedures that result in: (i) very high costs of therapy of up to half of a million dollars; (ii) delays of weeks or months to infuse CAR-T cells to patients with rapidly progressing disease; and (iii) heterogeneous composition and terminal differentiation of infused CAR-T cells as a result of ex vivo culture that limit CAR-T cell engraftment and persistence. Effort to overcome these limitations have focused on closed and automatic manufacturing devices to contain the labor needed to manufacture CAR-T cells ex vivo, and allogeneic off-the-shelf CAR-T cells have been proposed to overcome the need of CAR-T cell manufacturing for each single patient. Despite significant achievements in this space, reducing the time, costs and regulatory burden remains a deep unmet need in CAR-T cell therapy and significant reducing or eliminating ex vivo procedures remains a critical unmet need. In vivo generation of CAR-T cells would eliminate the need for ex vivo procedures, prevent the terminal differentiation of ex vivo expanded CAR-T cells and ensure the potency and longevity of autologous T cells as compared to allogeneic CAR-T cell products that are extensively manipulated to prevent rejection and graft- versus-host disease The research outlined in this proposal develops new biomaterials approaches to reduce the time and effort to produce CAR-T cells in vitro, to enhance CAR-T cell efficacy and persistence in vivo and, finally, to eliminate ex vivo manipulation entirely by generating CAR-T cells entirely within the patient. We propose that biomaterial scaffolds displaying anti-CD3/CD28 antibodies and releasing pro-proliferative interleukins will mediate simultaneous activation and viral transduction of T cells without centrifugation (spinoculation) or transduction agents (retronectin, polybrene) and will facilitate ex vivo genetic reprogramming of T cells by reducing the time and expense of activating naive T-cells and transducing them with viral vectors. We next propose that directly implanting scaffolds seeded with peripheral blood mononuclear cells and CAR- encoding viral vectors will promote release of CAR-T cells into circulation, eliminating ex vivo CAR-T isolation and proliferation protocols to promote a less differentiated cell phenotype associated with longer in vivo persistence. Finally, we propose that, through the inclusion of encapsulated T-cell attracting cytokines, implanted biomaterial scaffolds will generate CAR-T cells entirely in situ through recruitment of host T cells to the scaffold, in-scaffold reprogramming of recruited T cells with resident CAR-encoding viral vectors, and release of reprogrammed CAR-T cells. We expect that our results will provide a basis for a general cellular therapeutic strategy and promote widespread patient access. In addition to the obvious applications in blood cancers, this rational materials-based approach for cellular manufacturing will be adopted to program therapeutic lymphocytes in solid tumors and for other diseases.

项目概要尽管嵌合抗原受体（CAR）-T 细胞疗法在治疗肿瘤方面取得了前所未有的临床成功，广泛的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) 治疗费用非常高，高达 50 万美元； (ii) 延迟数周或数月的时间来输注 CAR-T 为疾病快速进展的患者提供细胞； (iii) 异质成分和终端离体培养导致输注的 CAR-T 细胞分化，限制了 CAR-T 细胞的植入和坚持。克服这些限制的努力集中在封闭式自动化制造设备上包含离体制造 CAR-T 细胞所需的劳动力，同种异体现成 CAR-T 细胞旨在克服为每位患者制造 CAR-T 细胞的需求。尽管显着尽管在这一领域取得了成就，但减少时间、成本和监管负担仍然是一个未得到满足的深刻需求 CAR-T 细胞疗法和显着减少或消除离体程序仍然是一个未满足的关键需求。在体内生成 CAR-T 细胞将消除离体程序的需要，防止最终体外扩增 CAR-T 细胞的分化，并确保自体 T 细胞的效力和寿命与经过广泛操作以防止排斥和移植的同种异体 CAR-T 细胞产品相比该提案中概述的研究开发了新的生物材料方法来减少抗宿主病体外生产 CAR-T 细胞的时间和精力，以增强 CAR-T 细胞在体内的功效和持久性，最后，通过完全在患者体内生成 CAR-T 细胞来完全消除离体操作。我们提出生物材料支架展示抗 CD3/CD28 抗体并释放促增殖细胞白细胞介素将介导 T 细胞同时激活和病毒转导，无需离心（纺丝）或转导剂（retronectin、polybrene），将促进离体遗传重编程通过减少激活初始 T 细胞并用病毒载体转导它们的时间和费用来增强 T 细胞的活性。接下来我们建议直接植入接种有外周血单核细胞和CAR-的支架编码病毒载体将促进 CAR-T 细胞释放到循环中，消除离体 CAR-T 分离和增殖方案，以促进与体内较长时间相关的分化程度较低的细胞表型坚持。最后，我们建议，通过包含封装的 T 细胞吸引细胞因子，植入的生物材料支架将通过招募宿主 T 细胞来完全原位生成 CAR-T 细胞支架，用驻留的 CAR 编码病毒载体对招募的 T 细胞进行支架内重新编程，以及释放重编程的 CAR-T 细胞。我们期望我们的结果将为一般细胞提供基础治疗策略并促进广泛的患者获取。除了在血液中的明显应用外癌症，这种基于合理材料的细胞制造方法将被用于编程实体瘤和其他疾病中的治疗性淋巴细胞。