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

• 批准号: 10686448
• 负责人: Chuntang Fu
• 金额: $ 100万
• 依托单位: DIAGNOLOGIX, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-03 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10686448
• 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; Mus; Organ; Outcome; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; Play; Population; Production; Proliferating; Protocols documentation; Regulatory T-Lymphocyte; Reporting; 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

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 需要更有必要的选择性和可持续疗法来治疗越来越多的免疫力 相关疾病。例如，自身免疫性疾病影响了曼联超过5000万患者 单独的州，在欧洲儿童中，欧洲儿童的1型糖尿病的年增加3.4％ 1989年和2013年。尽管常规治疗可能会缓解症状，但它们通常不那么具体，并且 需要长期药物。对使用监管t（TREG）开发“活着的药物”的兴趣越来越大 通过更好地了解免疫稳态的细胞，用于治疗各种免疫学相关疾病 以及最近采用自适应细胞疗法的临床结果。许多早期临床试验 用Treg细胞产品证明了这种方法的可行性和安全性。标准化Treg单元 制造业一直是一个重大挑战，涉及选择细胞源，净化方法， 工程以及扩展Treg细胞，产品规范和释放标准。过去几个 多年来，我们一直在开发基于微泡的方法来简化制造 热细胞生产的过程，包括T细胞选择，激活和工程。中的一个 Treg细胞制造的主要障碍是隔离可以大大扩展的高纯度细胞 用于临床疗法。要以高纯度分离treg细胞，使用一组表面标记的多参数分类 （例如CD4，CD25，CD127）。当前两种主要技术，荧光激活的细胞分选 （FACS）和磁细胞分类（MAC），仅一个人无法满足隔离大型GMP的需求 高纯度的Treg细胞。我们最近发明了一个迭代，有针对性的基于微泡的平台 对于多个参数细胞的分类，从刻度上的血液样本中分类。此外，我们还 证明抗CD3/CD28共轭微泡对无珠T细胞激活非常有效 和长期扩张。有了这个基础，我们将建立一个创新平台，用于Treg细胞处理， 并将证明产生足够高纯度的足够功能性Treg细胞的可行性 在第一阶段的血液样本。随后，我们将产生抗原特异性CAR（嵌合抗原受体） 符合II期临床规格的Treg细胞。一旦成功，这可能会加速 这种承诺疗法可以在固体器官或 造血干细胞移植以及与其他免疫相关疾病作斗争。