Toward GMP production of antigen specific regulatory T cells

致力于抗原特异性调节性 T 细胞的 GMP 生产

基本信息

批准号：
10324630
负责人：
Chuntang Fu
金额：
$ 30万
依托单位：
DIAGNOLOGIX, LLC
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：
2021-09-03 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10324630
关键词：
Adoption Adoptive Cell Transfers Adult Affect Antigen-Presenting Cells Antigens Autoimmune Diseases Autologous Biological Assay Blood Circulation Blood Component Removal Blood specimen CD28 gene CD3 Antigens CD4 Positive T Lymphocytes Cell Separation Cell Therapy Cell Transplantation Cells Child Clinical Disease Engineering Ensure European Fluorescence-Activated Cell Sorting Foundations Gold HLA-A2 Antigen Hematopoietic Stem Cell Transplantation Homeostasis Human IL2RA gene IL7R gene Immune Immunity Immunologics In Vitro Insulin-Dependent Diabetes Mellitus Lead Lentivirus Vector Lymphoid Tissue Magnetism Methods Microbubbles Modeling Organ Outcome Patients Peripheral Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Phase Play Population Production Proliferating Protocols documentation Regulatory T-Lymphocyte Reproducibility Retroviral Vector Role Self Tolerance Solid Sorting - Cell Movement Source Standardization Surface System T-Cell Activation T-Cell Development T-Lymphocyte Techniques Technology Therapeutic Thymus Gland Time United States Viral Virus base cellular transduction central tolerance chimeric antigen receptor clinical application clinical translation conventional therapy cost efficient early phase clinical trial gene therapy graft vs host disease improved in vivo innovation innovative technologies interest manufacturing process meetings microchip mouse model peripheral tolerance reduce symptoms response safety and feasibility skin xenograft tool transduction efficiency

项目摘要

ABSTRACT There is an unmet need for more selective and sustainable therapeutics to treat a growing number of immune- related diseases. For example, autoimmune disorders affect more than 50 million patients in the United States alone and there was a 3.4% annual increase in type 1 diabetes among European children between 1989 and 2013. While conventional treatments may alleviate symptoms, they are often less specific and require long-term medication. There is a growing interest in developing “living drugs” with regulatory T (Treg) cells for treating various immunity-related diseases, given better understanding of immunological homeostasis and recent promising clinical outcomes from applying adoptive cell therapies. Many early-phase clinical trials with Treg cell products have demonstrated the feasibility and safety of this approach. Standardizing Treg cell manufacturing has been a substantial challenge, involving choice of cell source, methods for purifying, engineering, and expanding Treg cells, product specification, and release criteria. Over the past several years, we have been developing targeted microbubble-based methods for streamlining the manufacturing processes for therapeutic cell production, including T cell selection, activation, and engineering. One of the major hurdles for Treg cell manufacturing is isolating high purity cells that can be greatly expanded at scale for clinical therapy. To isolate Treg cells in high purity, multiparametric sorting using a set of surface markers (e.g. CD4, CD25, CD127) is needed. The two current major techniques, fluorescence-activated cell sorting (FACS) and magnetic cell sorting (MACS), alone cannot meet the demand for isolating large-scale GMP grade Treg cells of high purity. We have recently invented an iterative, targeted microbubble-based platform for multiple parametric cell sorting at scale from apheresis blood samples. In addition, we have also demonstrated that anti-CD3/CD28 conjugated microbubbles are very efficient for bead-free T cell activation and long-term expansion. With this foundation, we will build an innovative platform for Treg cell processing, and will demonstrate the feasibility of producing adequate functional Treg cells of high purity from apheresis blood samples in phase I. Subsequently, we will generate antigen-specific CAR (chimeric antigen receptor) Treg cells that meet clinical specifications in phase II. Once successful, this could accelerate the adoption of this promising therapy to accomplish durable responses of suppressing rejection following solid organ or hematopoietic stem cell transplantation, as well as combating other immune-related disorders.

抽象的对于更具选择性和可持续的治疗方法来治疗越来越多的免疫性疾病的需求尚未得到满足。例如，自身免疫性疾病影响着美国超过 5000 万患者。仅在各州，欧洲儿童中 1 型糖尿病的年增长率为 3.4% 1989 年和 2013 年。虽然传统治疗可能会缓解症状，但它们往往不太具体，而且效果不佳。人们对开发具有调节性 T (Treg) 的“活药物”越来越感兴趣。更好地了解免疫稳态，用于治疗各种免疫相关疾病的细胞以及最近应用过继细胞疗法取得的有希望的临床结果。 Treg细胞产品已经证明了这种标准化Treg细胞方法的可行性和安全性。制造一直是一个巨大的挑战，涉及细胞来源的选择、纯化方法、工程，并扩展 Treg 细胞、产品规格和发布标准。多年来，我们一直在开发基于微泡的有针对性的方法来简化制造治疗性细胞生产过程，包括 T 细胞选择、激活和改造。 Treg 细胞制造的主要障碍是分离可以大规模扩展的高纯度细胞使用一组表面标记以高纯度、多参数分选分离 Treg 细胞用于临床治疗。（例如 CD4、CD25、CD127）是当前的两种主要技术，荧光激活细胞分选。单独使用流式细胞仪（FACS）和磁性细胞分选仪（MACS）无法满足大规模GMP分离的需求我们最近发明了一种基于微泡的迭代、靶向平台。用于从单采血液样本中进行大规模的多参数细胞分选。证明抗 CD3/CD28 缀合微泡对于无珠 T 细胞激活非常有效在此基础上，我们将建立一个Treg细胞加工的创新平台，并将证明通过血浆分离术生产足够的高纯度功能性 Treg 细胞的可行性第一阶段的血液样本。随后，我们将生成抗原特异性的CAR（嵌合抗原受体）符合 II 期临床规范的 Treg 细胞一旦成功，可能会加速采用。这种有前途的疗法可以实现抑制实体器官或器官排斥反应的持久反应造血干细胞移植，以及对抗其他免疫相关疾病。