Engineering stem cell-derived T cells for immunotherapy

用于免疫治疗的工程干细胞衍生 T 细胞

基本信息

批准号：
9911932
负责人：
Patrick C Chang
金额：
$ 3.64万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9911932
关键词：
3-Dimensional Address Adoptive Cell Transfers Allogenic Antigens Autologous Biological Biological Assay Blood CD8B1 gene CRISPR/Cas technology Cell Line Cell Therapy Cells Cellular immunotherapy Clinical Complex Disease Engineering FDA approved Flow Cytometry Generations Genes Genetic Recombination Genetic Transcription Goals Guide RNA HLA-A gene Hematopoietic stem cells Homologous Transplantation Human Human Engineering Immune system Immunotherapy In Vitro Knock-out Lymphocyte Count MHC Class I Genes Mediating Methods Mus Organoids Patients Process RAG1 gene Refractory Disease Reproducibility Serum Source Specificity System T cell differentiation T-Cell Development T-Cell Receptor T-Lymphocyte Techniques Thymus Gland Time TimeLine Tumor Antigens V(D)J Recombination antigen-specific T cells arm beta-2 Microglobulin cancer cell cancer immunotherapy cell type cellular engineering cellular transduction chimeric antigen receptor cost design engineered T cells engineered stem cells experimental study genome-wide graft vs host disease human pluripotent stem cell improved in silico in vitro activity in vivo individualized medicine insertion/deletion mutation manufacturing process neoplastic cell next generation novel novel strategies novel therapeutics outcome forecast prevent receptor expression receptor-mediated signaling response screening self-renewal stem cells tumor

项目摘要

PROJECT SUMMARY/ABSTRACT Adoptive cell transfer (ACT) immunotherapies repurpose T cells from patients to specifically eliminate tumor cells by engineering them to express antigen-specific T cell receptors (TCRs) and chimeric antigen receptors (CARs). Currently, engineered T cell manufacturing is patient-specific and the timeline and cost of manufacture which may limit access to these new therapies. Some patients have a prognosis for survival that does not end beyond the inherent time required for cell manufacturing (on average 22 days for a current FDA-approved CAR- T product), and clinical responses are uneven and are related, at least partly, to the functional variability of the T cell composition of each product. This proposal seeks to remove these barriers by engineering human pluripotent stem cells (hPSCs) to generate non-alloreactive, antigen-specific mature, naïve T cells using the Artificial Thymic Organoid (ATO) system, a novel in vitro method for efficiently and reproducibly producing mature, naïve T cells from stem cell sources. CRISPR/Cas9 gene-editing will be used in hPSCs to generate insertion deletion mutations (INDELs) to knockout the beta-2-microglublin (B2M) gene, the critical subunit of all Class I MHC heterodimers, to prevent Class I MHC-mediated allo-rejection of T cells. To prevent the rearrangement of endogenous, and potentially allo-reactive TCRs, a multiplex CRISPR/Cas9 strategy will be used in hPSCs to completely excise the locus containing the recombination-activating-genes RAG1 and RAG2 in hPSCs. Together, these experiments will develop a platform for generating stem cell-derived, non-alloreactive mature naïve T cells that could ultimately be applied toward an “off-the-shelf” approach to T cell immunotherapy.

项目概要/摘要过继细胞转移 (ACT) 免疫疗法重新利用患者的 T 细胞来特异性消除肿瘤通过改造细胞来表达抗原特异性 T 细胞受体 (TCR) 和嵌合抗原受体目前，工程 T 细胞的制造是针对特定患者的，并且制造的时间线和成本也不同。这可能会限制一些患者的生存预后。超出了细胞制造所需的固有时间（当前 FDA 批准的 CAR 平均需要 22 天） T 产品），并且临床反应不均匀，并且至少部分地与该提案旨在通过改造人体来消除这些障碍。多能干细胞 (hPSC) 使用人工胸腺类器官（ATO）系统，一种高效、可重复生产的新型体外方法来自干细胞来源的成熟、幼稚 T 细胞将用于 hPSC 中生成。插入缺失突变（INDEL）可敲除β-2-微球蛋白（B2M）基因，该基因是所有基因的关键亚基 I 类 MHC 异二聚体，可防止 I 类 MHC 介导的 T 细胞同种异体排斥。内源性和潜在同种异体反应性 TCR 的重排，多重 CRISPR/Cas9 策略将是用于 hPSC 中以完全切除含有重组激活基因 RAG1 和 RAG2 的基因座这些实验将共同开发一个用于产生干细胞衍生的、非同种异体反应性的平台。成熟的初始 T 细胞最终可用于 T 细胞免疫治疗的“现成”方法。