Effects of PGE2 on Reconstitution of Hematopoiesis and Immunity after UCBT

PGE2对UCBT后造血和免疫重建的影响

• 批准号: 8985871
• 负责人: VASSILIKI A BOUSSIOTIS
• 金额: $ 37.47万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-17 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8985871
• 关键词: Address; Affect; Antigens; Binding; Bioenergetics; Biological Models; CD8-Positive T-Lymphocytes; CD8B1 gene; Carnitine O-Palmitoyltransferase; Cell Differentiation process; Cell Therapy; Cells; Clinical; Cytomegalovirus; Development; Dimethyl Sulfoxide; Dinoprostone; Effector Cell; Employee Strikes; Enzymes; Epitopes; Fatty Acids; Funding; Gene Expression; Gene Expression Microarray Analysis; Generations; Genes; Glucose Transporter; Glycolysis; HLA A*0201 antigen; Hematopoiesis; Human; Human Herpesvirus 4; Immune; Immunity; In Vitro; Incidence; Individual; Life; Lipids; Mediating; Memory; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Mus; NOD/SCID mouse; Pathway interactions; Phenotype; Property; Relapse; Solid Neoplasm; System; T cell differentiation; T memory cell; T-Lymphocyte; Transgenic Mice; Transgenic Organisms; Tumor Immunity; Umbilical Cord Blood Transplantation; Up-Regulation; WT1 gene; acyl-CoA oxidase; base; fatty acid oxidation; fatty acid transport; glucose uptake; gp100 Antigen; imprint; improved; improved functioning; in vivo; leukemia; leukemia treatment; lipid metabolism; melanoma; mortality; neoplasm immunotherapy; novel; novel strategies; oxidation; pathogen; programs; promoter; public health relevance; reconstitution; transcription factor; tumor

 DESCRIPTION (provided by applicant): The present revision application is based on the original RO1 application entitled "Effects of PGE2 on reconstitution of hematopoiesis and immunity after umbilical cord blood transplantation (UCBT)" (R01 CA183605-01). During the past 15 months we performed studies to investigate the effects of PGE2 in T cells. Using microarray gene expression analysis we determined that a number of genes were selectively altered by PGE2 in UCB T cells. The most striking effect was detected on the expression of the transcription factor NR4A2. NR4A transcription factors have recently emerged as regulators of metabolic function. Using ChIP-PCR, we found that PGE2-induced NR4A2 could bind on the promoter of carnitine palmitoyltransferase (CPT1A), the rate limiting enzyme of fatty acid transport to the mitochondria, resulting in upregulation of CPT1A and increased rate of fatty acid oxidation (FAO) in T cells. PGE2 also diminished the expression of the glucose transporter Glut 1 and the uptake of glucose and this effect was more prominent in CD8+ T cells. Using antigen-specific mouse CD8+ T cells and a model system of in vitro differentiation of T effector and T memory cells, we found that PGE2 promoted T memory cell differentiation. We also determined that PGE2 treatment of the UCB grafts had a significant clinical impact on pathogen-specific and tumor-specific immunity. Compared to control UCBT recipients, PGE2-UCBT recipients had significantly reduced incidence of EBV and CMV reactivation, and reduced incidence of leukemia relapse. Divergence in the metabolic reprogramming of T cells is critical to effectively imprint distinct T cell fates. It has been shown that metabolic switch to FAO promotes the conversion of CD8+ T effector to long-lived T memory cells. Enforcing FAO in various T cell experimental systems induced increased numbers of memory CD8+ T cells and resulted in the generation of memory precursors, TCM and TSCM cells, which display enhanced anti-tumor function. Taken together our in vitro findings on the effects of PGE2 on T cell metabolic reprogramming and T memory cell development, and our clinical observations of improved immunity in PGE2-UCBT recipients suggest that PGE2 treatment might improve anti-tumor immunity by mediating metabolic reprogramming of tumor-specific T cells. This approach might represent a novel method to generate potent tumor-specific T cells for cell based tumor immunotherapy. To address this hypothesis in this revision application we will pursue the following specific aim to: SA1: Determine the mechanisms of PGE2-mediated metabolic reprogramming and its effects on the generation and function of tumor-specific T cells. A) To determine the effects of PGE2 on the molecular, metabolic and gene expression programs of tumor- specific mouse T cells and on their anti-tumor function in vivo. B) To determine the effects of PGE2 on the molecular and metabolic and functional properties of tumor-specific human T cells.

 说明（申请人提供）：本修订申请基于原RO1申请，题为“PGE2对脐带血移植（UCBT）后造血和免疫重建的影响”（R01 CA183605-01）。在过去 15 个月中，我们进行了研究来调查 PGE2 对 T 细胞的影响。通过微阵列基因表达分析，我们确定 UCB T 细胞中的许多基因被 PGE2 选择性改变。最显着的影响是在转录因子 NR4A2 的表达上检测到的。 NR4A 转录因子最近已成为代谢功能的调节因子。使用 ChIP-PCR，我们发现 PGE2 诱导的 NR4A2 可以与肉碱棕榈酰转移酶 (CPT1A) 的启动子结合，CPT1A 是脂肪酸转运到线粒体的限速酶，导致 CPT1A 上调并增加 T 细胞中脂肪酸氧化 (FAO) 的速率。 PGE2 还减少了葡萄糖转运蛋白 Glut 1 的表达和葡萄糖的摄取，这种效应在 CD8+ T 细胞中更为突出。使用抗原特异性小鼠 CD8+ T 细胞以及 T 效应细胞和 T 记忆细胞体外分化模型系统，我们发现 PGE2 促进 T 记忆细胞分化。我们还确定 UCB 移植物的 PGE2 治疗对病原体特异性和肿瘤特异性免疫具有显着的临床影响。与对照 UCBT 接受者相比，PGE2-UCBT 接受者的 EBV 和 CMV 再激活发生率显着降低，白血病复发率也降低。 T 细胞代谢重编程的差异对于有效地印记不同的 T 细胞命运至关重要。研究表明，代谢转变为FAO可促进CD8+ T效应细胞向长寿命T记忆细胞的转化。在各种T细胞实验系统中执行FAO可诱导记忆CD8+ T细胞数量增加，并导致记忆前体细胞、TCM和TSCM细胞的产生，这些细胞表现出增强的抗肿瘤功能。综合我们关于 PGE2 对 T 细胞代谢重编程和 T 记忆细胞发育影响的体外研究结果，以及我们对 PGE2-UCBT 接受者免疫力改善的临床观察，表明 PGE2 治疗可能通过介导肿瘤特异性 T 细胞的代谢重编程来改善抗肿瘤免疫力。这种方法可能代表了一种生成有效的肿瘤特异性 T 细胞用于基于细胞的肿瘤免疫治疗的新方法。为了在本次修订申请中解决这一假设，我们将追求以下具体目标： SA1：确定 PGE2 介导的代谢重编程的机制及其对肿瘤特异性 T 细胞的生成和功能的影响。 A) 确定PGE2对肿瘤特异性小鼠T细胞的分子、代谢和基因表达程序及其体内抗肿瘤功能的影响。 B) 确定 PGE2 对肿瘤特异性人类 T 细胞的分子、代谢和功能特性的影响。