Engineering detours around the biologic barriers to allogeneic, iPSC-derived CAR T immunotherapy

工程绕开了同种异体、iPSC 衍生的 CAR T 免疫疗法的生物障碍

基本信息

批准号：
10607952
负责人：
Sang Pil Yoo
金额：
$ 4.25万
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：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10607952
关键词：
3&apos Untranslated Regions 3-Dimensional Address Affect Allogenic Antigens Autologous Biological CAR T cell therapy CD19 gene CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene CRISPR/Cas technology Cell Differentiation process Cell Maturation Cell Therapy Cells Cellular biology Cellular immunotherapy Clinical Data Development Engineering Excision Generations Genes Genetic Genetic Transcription Granzyme Health Services Accessibility Hematologic Neoplasms Immunoglobulin Constant Region Immunotherapy In Vitro Lymphoid Malignant Neoplasms Mature T-Lymphocyte Methods Modeling Organoids Pathway interactions Patients Pattern Physiological Pluripotent Stem Cells Positioning Attribute Rag1 Mouse Receptor Cell Receptor Signaling Refractory Research Risk Signal Transduction Source System T cell differentiation T cell therapy T-Cell Development T-Cell Receptor T-Lymphocyte Technology Thymus Gland Transgenes Tumor Antigens cancer type chimeric antigen receptor chimeric antigen receptor T cells clinical care constitutive expression cost cytotoxicity graft vs host disease human pluripotent stem cell improved in vivo individualized medicine induced pluripotent stem cell innovation interest manufacturing process monolayer multipotent cell novel prevent receptor expression receptor-mediated signaling response stem cell technology stem cells transcriptome sequencing

3&apos Untranslated Regions 3-Dimensional Address Affect Allogenic Antigens Autologous Biological CAR T cell therapy CD19 gene CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene CRISPR/Cas technology Cell Differentiation process Cell Maturation Cell Therapy Cells Cellular biology Cellular immunotherapy Clinical Data Development Engineering Excision Generations Genes Genetic Genetic Transcription Granzyme Health Services Accessibility Hematologic Neoplasms Immunoglobulin Constant Region Immunotherapy In Vitro Lymphoid Malignant Neoplasms Mature T-Lymphocyte Methods Modeling Organoids Pathway interactions Patients Pattern Physiological Pluripotent Stem Cells Positioning Attribute Rag1 Mouse Receptor Cell Receptor Signaling Refractory Research Risk Signal Transduction Source System T cell differentiation T cell therapy T-Cell Development T-Cell Receptor T-Lymphocyte Technology Thymus Gland Transgenes Tumor Antigens cancer type chimeric antigen receptor chimeric antigen receptor T cells clinical care constitutive expression cost cytotoxicity graft vs host disease human pluripotent stem cell improved in vivo individualized medicine induced pluripotent stem cell innovation interest manufacturing process monolayer multipotent cell novel prevent receptor expression receptor-mediated signaling response stem cell technology stem cells transcriptome sequencing

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项目摘要

PROJECT SUMMARY/ABSTRACT Chimeric antigen receptor (CAR) T cell therapy has produced remarkable results in otherwise treatment- refractory hematological malignancies. Currently, the process of manufacturing autologous CAR T cells is challenging due to the need for de novo generation of each individualized therapy and the inherent variability in T cell biology between patients, leading to unpredictable and inconsistent clinical responses. As a result, there is a growing interest in generating CAR T cells in vitro from an infinitely renewing, allogeneic source of human pluripotent stem cells (hPSCs). One advantage of this approach is that hPSCs are highly amenable to genetic editing, providing multiple avenues to manipulate the function of the final T cell product. However, there are several biologic barriers to generating CAR T cells from hPSCs. First, expression of the CAR transgene early in T cell differentiation diverts development towards the innate lymphoid pathway instead of the conventional T lineage. Second, the removal of the endogenous T cell receptor (TCR) to prevent alloreactivity leads to a block in development, as maturing T cells can no longer undergo positive selection. This proposal seeks to overcome these challenges to generating non-alloreactive, stem cell-derived CAR T cells by deploying an innovative stage-specific expression strategy and a novel in vitro method to induce the differentiation of mature conventional T cells, the Artificial Thymic Organoid (ATO) system. The ATO is a first-in-class, in vitro method for efficiently generating mature, single positive (SP) CD8+ and CD4+ T cells from multiple stem cell sources, including hPSCs. Improving upon previously established systems, the ATO can robustly support positive selection and maturation to the SP stage. Our preliminary studies have already demonstrated that innate fate diversion can be overcome by achieving delayed, stage-specific expression of the CAR transgene that is limited to mature T cells. In this proposal, Specific Aim 1 will build upon the delayed CAR expression model and evaluate how the disruption of the endogenous TCR affects CAR T cell development in the ATOs. Specific Aim 2 will evaluate two different methods of delivering exogenous positive selection signals to rescue development in the absence of the endogenous TCR. These methods include expression of a tonically signaling CAR or transiently expressing an exogenous TCR at the physiologically appropriate double positive (DP) T cell stage. The approaches developed in this proposal will enable the generation of hPSC-derived, non-alloreactive T cell therapy, ultimately reducing the cost and increasing the access to treatment for more patients in need.

项目摘要/摘要嵌合抗原受体（CAR）T细胞疗法在其他治疗方面产生了显着的结果 - 难治性血液学恶性肿瘤。目前，制造自体汽车T细胞的过程是由于需要从从头生成每种个性化疗法以及固有的可变性，因此具有挑战性患者之间的T细胞生物学，导致不可预测和不一致的临床反应。结果，对从无限更新，人类的同种异体来源产生汽车T细胞的兴趣日益兴趣多能干细胞（HPSC）。这种方法的一个优点是HPSC高度适合遗传编辑，提供多种途径来操纵最终T细胞产品的功能。但是，从HPSC产生CAR T细胞有几个生物学障碍。首先，表达 T细胞分化早期的CAR转基因将发育转移到先天淋巴样途径而不是传统的T谱系。其次，去除内源性T细胞受体（TCR）以防止同种反应性会导致发育的障碍，因为成熟的T细胞不再可以接受阳性选择。该提案旨在克服这些挑战，以产生非异化性，干细胞衍生的汽车通过部署创新阶段特异性表达策略和一种新颖的体外方法来诱导细胞成熟常规T细胞的分化，人造胸腺类器官（ATO）系统。 ATO是一种一流的体外方法，用于有效产生成熟的单个正（SP）CD8+ 来自包括HPSC在内的多个干细胞源的CD4+ T细胞。改进先前建立的系统，ATO可以强大地支持阳性选择和成熟到SP阶段。我们的初步研究已经证明，可以通过实现延迟的，特定于阶段的特定阶段来克服先天的命运转移限于成熟T细胞的CAR转基因的表达。在此提案中，特定目标1将基于延迟的汽车表达模型并评估内源性TCR的破坏如何影响CAR T细胞 Atos的发展。特定目标2将评估两种交付外源阳性的不同方法在没有内源性TCR的情况下，选择信号以营救发育。这些方法包括在生理上表达了音调信号汽车或瞬时表达外源TCR的表达适当的双阳性（DP）T细胞阶段。本提案中开发的方法将使 HPSC衍生的非异化性T细胞疗法的产生，最终降低了成本并增加获得更多有需要的患者的治疗。