Functional T cells from iPSCs Using Chimeric Antigen Receptors

项目摘要： T细胞免疫疗法有望成为未来医学的主要参与者。CD 19 CAR T的成功 急性淋巴细胞白血病的细胞已经给人留下了深刻的印象，并让我们看到了下一代的细胞。 工程化T细胞可能有能力。在未来，CAR靶向T细胞可能是有价值的免疫疗法， 不仅仅是癌症特别地，靶向调节性T细胞疗法是治疗恶性肿瘤的有希望的可能性。 在移植受体中的自身免疫性病症和耐受诱导。CAR T细胞的当前方法 生产依赖于在每个患者的基础上费力的分离、修饰和扩增步骤。 这使得它们非常昂贵，几乎没有现成产品的可能性。诱导多能干 iPSC细胞（iPSC）允许将成年体细胞重编程为来自特定疾病患者的干细胞样命运。 MHC单倍型来克服这个问题。MHC库iPSC和CAR技术集的组合 可再生且高度可控的现成T细胞治疗剂的阶段。然而，努力 将iPSC分化为T细胞一直在努力产生具有强大T细胞的造血祖细胞， 细胞潜能和在CD 4/8双阳性阶段阻断T细胞成熟。在本提案中，我们 证实了调节NOTCH信号通路对T-能力祖细胞输出的显著影响。 我们建议将这一成功与单细胞RNAseq结合联合收割机来揭示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。它还将提高分化培养的效率。在 此外，我们建议在免疫过程中使用具有一系列抗原亲和力的汽车的时间控制表达。 T细胞分化以绕过其发育阻滞并驱动成熟为功能性T细胞。 这两个协同但不依赖的目标将是迈向真正功能性iPSC的重要而有力的一步。 也将为探索驱动CD 4辅助性T细胞所需的信号打开大门 承诺和T reg血统规范。

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.