Functional T cells from iPSCs Using Chimeric Antigen Receptors

使用嵌合抗原受体从 iPSC 中提取功能性 T 细胞

• 批准号: 9760329
• 负责人: Dar Heinze
• 金额: $ 6.77万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9760329
• 关键词: Acute Lymphocytic Leukemia; Adult; Affinity; Antigens; Area; Autoimmune Process; Biological Assay; Bypass; CD19 gene; CD34 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Cell Lineage; Cell Maturation; Cell Therapy; Cells; Clinical; Cues; Cytotoxic T-Lymphocytes; Data; Defect; Derivation procedure; Development; Doxycycline; Endothelial Cells; Endothelium; Fellowship; Future; Goals; Haplotypes; Hematopoietic stem cells; Immunotherapy; Malignant Neoplasms; Mature T-Lymphocyte; Mediating; Mediator of activation protein; Medicine; Methods; Modification; Molecular Profiling; Mus; Nature; Notch Signaling Pathway; Output; Patients; Phase; Population; Production; Protocols documentation; Publishing; Regulatory T-Lymphocyte; Reporting; Signal Pathway; Signal Transduction; Somatic Cell; Specificity; Stem cells; Surface; T cell differentiation; T-Cell Development; T-Lymphocyte; T-Lymphocyte Subsets; T-cell receptor repertoire; Technology; Tetanus Helper Peptide; Tetracyclines; Therapeutic; Time; Transfusion; Transplant Recipients; Tumor stage; Umbilical Cord Blood; Work; base; chimeric antigen receptor; chimeric antigen receptor T cells; dosage; engineered T cells; hematopoietic differentiation; improved; induced pluripotent stem cell; next generation; novel; progenitor; promoter; receptor; receptor expression; single cell sequencing; stem-like cell; success; transcriptome sequencing

Project Summary: T cell immunotherapy is poised to be a major player in the future of medicine. The success of CD19 CAR T cells in acute lymphoblastic leukemia has been impressive and gives a glimpse of what the next generation of engineered T cells may be capable of. In the future, CAR-targeted T cells may be valuable immunotherapy for more than just cancer. In particular, targeted T regulatory cell therapy is a promising possibility for treatment of autoimmune conditions and tolerance induction in transplant recipients. Current methods of CAR T cell production are dependent on laborious isolation, modification, and expansion steps on a per-patient basis. This makes them highly expensive with little possibility of an off-the-shelf product. Induced pluripotent stem cells (iPSC) allow the reprogramming of adult somatic cells to a stem-cell like fate from patients with specific MHC haplotypes to overcome this issue. The combination of MHC-banked iPSCs and CAR technology sets the stage for a renewable and highly controllable off-the-shelf T cell therapeutic. However, efforts to differentiate iPSCs into T cells have struggled with the production of hematopoietic progenitors with robust T cell potential and a blockade in T cell maturation at the CD4/8 double positive stage. In this proposal we demonstrate the marked impact of modulating the NOTCH signaling pathway on T-capable progenitor output. We propose to combine this success with single-cell RNAseq to reveal the true identity of the T cell progenitor. This work will uncover key signaling pathways and answer lingering questions about the nature of iPSC- derived hematopoietic progenitors. It will also leverage improved efficiency in differentiation cultures. In addition, we propose to use temporally controlled expression of CARs with a range of antigen affinities during T cell differentiation to bypass their developmental blockade and drive maturation into functional T cells. These two synergistic but not dependent goals will be a vital and powerful step toward truly functional iPSC- derived T cells and would also open the door for exploring signals necessary to drive CD4 helper T cell commitment and T reg lineage specification.

项目概要： T 细胞免疫疗法有望成为未来医学的主要参与者。 CD19 CAR T的成功 急性淋巴细胞白血病细胞的研究令人印象深刻，让我们得以一睹下一代细胞的潜力 工程化 T 细胞可能有能力。未来，CAR靶向T细胞可能成为有价值的免疫疗法 不仅仅是癌症。特别是，靶向 T 调节细胞疗法是治疗以下疾病的有希望的可能性： 自身免疫性疾病和移植受者的耐受诱导。目前CAR-T细胞的方法 生产依赖于针对每个患者的费力隔离、修改和扩展步骤。 这使得它们非常昂贵，几乎不可能成为现成产品。诱导多能干 细胞（iPSC）允许将成年体细胞重新编程为患有特定疾病的患者的干细胞样命运 MHC 单倍型可以克服这个问题。 MHC 库 iPSC 和 CAR 技术集的结合 可再生且高度可控的现成 T 细胞疗法的舞台。然而，努力 iPSC 分化为 T 细胞一直在努力生产具有强大 T 细胞的造血祖细胞 细胞潜能和 T 细胞成熟在 CD4/8 双阳性阶段的阻断。在这个提案中我们 证明了调节 NOTCH 信号通路对 T 能力祖细胞输出的显着影响。 我们建议将这一成功与单细胞 RNAseq 结合起来，以揭示 T 细胞祖细胞的真实身份。 这项工作将揭示关键的信号通路并回答有关 iPSC 性质的挥之不去的问题。 衍生的造血祖细胞。它还将利用差异化培养中提高的效率。在 此外，我们建议在不同阶段使用具有一系列抗原亲和力的临时控制的 CAR 表达。 T 细胞分化以绕过其发育障碍并促进成熟为功能性 T 细胞。 这两个协同但不相互依赖的目标将是迈向真正功能性 iPSC 的重要而有力的一步。 衍生的 T 细胞，也将为探索驱动 CD4 辅助 T 细胞所需的信号打开大门 承诺和 T reg 谱系规范。