Delineating mechanisms underlying the enhanced stability and functionality of CD2-KO Tregs and chimeric antigen receptor (CAR) Tregs and their application in xenotransplantation

描述 CD2-KO Tregs 和嵌合抗原受体 (CAR) Tregs 稳定性和功能增强的机制及其在异种移植中的应用

• 批准号: 10646753
• 负责人: Mohsen Khosravi Maharlooei
• 金额: $ 1.38万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-17 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10646753
• 关键词: Acetyl Coenzyme A; Acetylation; Address; Adverse effects; Antibodies; Antigens; Autoimmunity; CD2 Antigens; Cells; Cellular immunotherapy; Citrates; Citric Acid Cycle; Cytosol; Data; Deacetylase; Development; Equilibrium; Excision; FOXP3 gene; Family suidae; Genetic Transcription; Genome; Glycolysis; HLA Antigens; Histone Acetylation; Human; Immune; Immune response; Immune system; Immunosuppression; In Vitro; Inflammatory; Islets of Langerhans Transplantation; Link; Mediating; Metabolic; Metabolism; Mitochondria; Modeling; NADH; Organ; Organ Transplantation; Pathway interactions; Post-Translational Protein Processing; Protein Acetylation; Proteins; Regimen; Regulatory T-Lymphocyte; Resistance; Respiration; SIRT1 gene; Signal Pathway; Signal Transduction; Siplizumab; Source; T cell therapy; T-Cell Depletion; T-Lymphocyte; Transplantation; Ubiquitin; Work; Xenograft procedure; cadmium ion; chimeric antigen receptor; clinical application; conditioning; effector T cell; efficacy evaluation; epigenome; histone modification; humanized mouse; immunoregulation; in vivo; islet; mouse model; prevent; transcriptome

PROJECT SUMMARY Regulatory T cell (Treg) therapy has been widely investigated for control of immune responses in different models of autoimmunity and transplantation. Despite promising results in using Tregs and chimeric antigen receptor (CAR) Tregs for tolerance induction, there are still major concerns regarding the plasticity of Tregs and the effect of T cell-depleting conditioning regimens on these cells that need to be addressed before their clinical application. We have discovered that CD2-KO Tregs and CAR Tregs are not only resistant to a conditioning regimen that includes anti-CD2 antibody (siplizumab), but also have increased FOXP3 expression and Treg functionality in vitro and in vivo. CD2 is a costimulatory molecule that is present in all T cells, including Tregs. Potential direct or indirect link between CD2 signaling and FOXP3 is not known. Our preliminary data show that CD2-KO Tregs have increased mTORc1 activity, glycolysis and mitochondrial respiration. We hypothesize that metabolically- active CD2-KO Tregs generate metabolites that through histone modification or post-translational modifications increase FOXP3 levels. In Aim 1, we will address these hypotheses. We will study CD2+ and CD2-KO Tregs at the genome, epigenome, transcriptome and protein levels and evaluate their metabolism and signaling pathways. In Aim 2, we will explore the translational aspects of this discovery in xenotransplantation setting. We have developed and validated a CAR against a universal pig antigen (class-I swine leukocyte antigen (SLA)) and have shown that SLA CAR Tregs harbor higher suppressive capacity in suppressing anti-pig immune responses in vitro as compared to polyclonal Tregs. We hypothesize that CD2-KO SLA CAR Tregs resist conversion to effector T cells and protect transplanted pig organs in an antigen-specific manner. We will evaluate the resistance of CD2-KO SLA CAR Tregs to conversion in inflammatory conditions and also the efficacy of CD2- KO SLA CAR Tregs in preventing the rejection of pig islets in a humanized mouse model of xeno-islet transplantation. Taken together, this work can potentially lead to the development of safer and more efficient Treg-based immunotherapies. Mechanistic studies could uncover molecules and/or signaling pathways involved in determining the fate/function of Tregs, targeting of which could alter the immune balance toward tolerance.

项目摘要 调节性T细胞（Treg）疗法已被广泛研究用于控制不同模型中的免疫应答 自身免疫和移植的关系尽管使用THBE和嵌合抗原受体 (CAR)尽管TdR用于耐受诱导，但仍存在关于TdR的可塑性和TdR的作用的主要关注点。 T细胞消耗调理方案对这些细胞，需要解决之前，他们的临床应用。 我们已经发现，CD 2-KO T细胞和CAR T细胞不仅对调节方案有抗性， 包括抗CD 2抗体（siplizumab），但也具有增加的FOXP 3表达和Treg功能， 体外和体内。CD 2是一种共刺激分子，存在于所有的T细胞中，包括T细胞。潜在直接 CD 2信号传导和FOXP 3之间的间接联系尚不清楚。我们的初步数据显示，CD 2-KO TcR 增加mTORc 1活性、糖酵解和线粒体呼吸。我们假设新陈代谢- 活性CD 2-KO TCLs通过组蛋白修饰或翻译后修饰产生代谢物， 增加FOXP 3水平。在目标1中，我们将讨论这些假设。我们将研究CD 2+和CD 2-KO T细胞， 基因组、表观基因组、转录组和蛋白质水平，并评估其代谢和信号传导 路径。在目标2中，我们将探讨这一发现在异种移植环境中的翻译方面。我们 已经开发并验证了针对通用猪抗原（I类猪白细胞抗原（SLA））的CAR 并且已经显示SLA CAR T细胞在抑制抗猪免疫中具有更高的抑制能力， 与多克隆TclA相比，体外反应。我们假设CD 2-KO SLA CAR T细胞抵抗 转化为效应T细胞并以抗原特异性方式保护移植的猪器官。我们将评估 CD 2-KO SLA CAR T细胞对炎症条件下转化的抗性以及CD 2-KO SLA CAR T细胞对炎症条件下转化的功效。 KO SLA CAR T细胞在异种胰岛人源化小鼠模型中预防猪胰岛排斥反应 移植总之，这项工作可能会导致开发更安全，更有效的 基于Treg的免疫疗法机制研究可以揭示参与的分子和/或信号通路 在确定TcB的命运/功能中，靶向TcB可以改变免疫平衡朝向耐受性。