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: 10543567
• 负责人: Chuntang Fu
• 金额: $ 10万
• 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 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10543567
• 关键词: Adoption; Adoptive Cell Transfers; Affect; Antigens; Autoimmune Diseases; Blood Component Removal; Blood specimen; CD28 gene; CD3 Antigens; Cell Separation; Cell Therapy; Cells; Child; Clinical; Disease; Engineering; European; Fluorescence-Activated Cell Sorting; Foundations; Hematopoietic Stem Cell Transplantation; Homeostasis; IL2RA gene; IL7R gene; Immune; Immunity; Immunologics; Insulin-Dependent Diabetes Mellitus; Lead; Magnetism; Methods; Microbubbles; Organ; Outcome; Patients; Pharmaceutical Preparations; Phase; Production; Regulatory T-Lymphocyte; Solid; Sorting - Cell Movement; Source; Standardization; Surface; T-Cell Activation; T-Lymphocyte; Techniques; Therapeutic; United States; base; chimeric antigen receptor; conventional therapy; early phase clinical trial; gene therapy; innovation; innovative technologies; interest; manufacturing process; reduce symptoms; response; safety and feasibility

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性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疾病。 相关疾病。例如，自身免疫性疾病影响着美国5000多万患者。 仅在美国，欧洲儿童1型糖尿病的年增长率为3.4%， 1989年和2013年。虽然常规治疗可以缓解症状，但它们往往不那么特异， 需要长期服药。人们对开发具有调节性T细胞（Treg）的“活性药物”的兴趣越来越大。 细胞治疗各种免疫相关疾病，给予更好的理解免疫稳态 以及最近应用过继细胞疗法的有希望的临床结果。许多早期临床试验 Treg细胞产品的研究已经证明了这种方法的可行性和安全性。标准化Treg细胞 制造是一个实质性的挑战，涉及细胞来源的选择，纯化方法， 工程化和扩增Treg细胞、产品质量标准和放行标准。过去几 多年来，我们一直在开发有针对性的基于微泡的方法，以简化制造过程。 用于治疗性细胞生产的方法，包括T细胞选择、活化和工程化。之一 Treg细胞制造的主要障碍是分离可以大规模扩增的高纯度细胞 用于临床治疗。为了以高纯度分离Treg细胞，使用一组表面标志物进行多参数分选 (e.g. CD4、CD25、CD127）。目前的两种主要技术，荧光激活细胞分选 流式细胞仪（FACS）和磁细胞分选（MACS），单独不能满足分离大规模GMP的需求 高纯度的分级Treg细胞。我们最近发明了一种迭代的、有针对性的基于微泡的平台， 用于从单采血液样品中大规模进行多参数细胞分选。我们亦 证明了抗CD 3/CD 28缀合的微泡对于无珠T细胞活化非常有效 长期扩张。在此基础上，我们将建立一个创新的Treg细胞处理平台， 并将证明从单采血液成分术产生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CAR（嵌合抗原受体） 满足II期临床规范的Treg细胞。一旦成功，这可能会加速采用 这种有前途的治疗方法，以实现持久的反应，抑制排斥反应后，实体器官或 造血干细胞移植，以及对抗其他免疫相关疾病。