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.

项目摘要： T细胞免疫疗法有望成为医学未来的主要参与者。 CD19车的成功 急性淋巴细胞白血病中的细胞令人印象深刻，并瞥见了下一代 工程T细胞可能能够。将来，靶向汽车的T细胞可能是有价值的免疫疗法 不仅仅是癌症。特别是，有针对性的T调节细胞疗法是治疗的有希望的可能性 移植受者的自身免疫性条件和耐受性诱导。当前汽车T细胞的方法 生产取决于艰苦的隔离，修改和扩展步骤。 这使它们变得非常昂贵，几乎没有现成的产品。诱导多能茎 细胞（IPSC）允许将成年体细胞重新编程为具有特定患者的患者的干细胞命运 MHC单倍型要克服这个问题。 MHC银行IPSC和汽车技术套装的组合 可再生且高度可控制的现成T细胞治疗的阶段。但是，努力 将IPSC区分为T细胞已经在造血祖细胞的产生中挣扎 在CD4/8双阳性阶段，细胞电位和T细胞成熟的封锁。在这个建议中，我们 证明调节Notch信号通路对T型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。 这项工作将发现关键的信号通路，并回答有关IPSC-性质的挥之不去的问题 衍生的造血祖细胞。它还将利用分化文化的提高效率。在 此外，我们建议在期间使用具有一系列抗原亲和力的汽车的时间控制的表达 T细胞分化以绕过它们的发育阻滞并将成熟驱动到功能性T细胞中。 这两个协同但不依赖的目标将是迈向真正功能型IPSC-的至关重要而有力的一步 派生的T细胞，还将为探索驱动CD4辅助T细胞所需的信号打开大门 承诺和t reg Lineage规范。