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.

摘要 对更具选择性和可持续的治疗方法的需求尚未得到满足，以治疗越来越多的免疫性- 相关疾病。例如，自身免疫性疾病影响了美国5000多万患者。 欧洲儿童中1型糖尿病的年增长率为3.4%。 1989年和2013年。虽然传统的治疗方法可能会缓解症状，但它们往往没有那么具体和 需要长期服药。人们对利用调节性T(Treg)开发“活性药物”的兴趣与日俱增 用于治疗各种免疫相关疾病的细胞，更好地了解免疫稳态 以及最近通过应用过继细胞疗法而获得的有希望的临床结果。许多早期临床试验 具有Treg细胞的产品已经证明了这种方法的可行性和安全性。标准化Treg细胞 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， 设计和扩展Treg细胞、产品规格和发布标准。在过去的几年中 多年来，我们一直在开发基于微气泡的有针对性的方法来简化制造 治疗细胞生产的过程，包括T细胞的选择、激活和工程化。其中一个 制造Treg细胞的主要障碍是分离出可大规模扩增的高纯度细胞 用于临床治疗。使用一组表面标记以高纯度、多参数分选分离Treg细胞 (例如，CD4、CD25、CD127)是必需的。目前的两种主要技术，荧光激活细胞分选 (FACS)和磁性细胞分选(MACS)，单独不能满足分离大规模GMP的需求 高纯度的级Treg细胞。我们最近发明了一种迭代的、基于微气泡的平台 用于从分离血液样本中大规模进行多参数细胞分选。此外，我们还 抗CD3/CD28偶联微泡对无珠化T细胞的激活作用 和长期扩张。在此基础上，我们将构建Treg细胞处理的创新平台， 并将证明从分离中生产足够的高纯度的功能性Treg细胞的可行性 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)。 第二阶段符合临床规格的Treg细胞。一旦成功，这可能会加速采用 这是一种有希望的疗法，可以实现持久的抑制固体器官或 造血干细胞移植，以及抗击其他免疫相关疾病。