DGK in T Cell Regulation and Tolerance

DGK 在 T 细胞调节和耐受中的作用

基本信息

批准号：
7740296
负责人：
THOMAS F GAJEWSKI
金额：
$ 39万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-21 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7740296
关键词：
Address Adenovirus Vector Adenoviruses Autoimmunity Binding Binding Sites Biological Assay Calcium Signaling Cell physiology Cells Chronic Critical Pathways Data Defect Dependence Development Diacylglycerol Kinase Disease Eating Effector Cell Event Family Gene Deletion Gene Expression Profiling Gene Targeting Genes Goals Immune Tolerance Immune system In Vitro Lead Ligation Lipids Luciferases Malignant Neoplasms Mediating Membrane Messenger RNA Molecular Mus Peripheral Phenotype Phosphorylation Phosphorylation Site Phosphotransferases Post-Translational Regulation Predisposition Principal Investigator Process Promoter Regions Property Regulation Reporter Resistance Role System T cell regulation T-Cell Activation T-Lymphocyte Tissues Transcriptional Regulation Transgenic Mice Transplantation Tolerance Tumor Immunity Tyrosine Tyrosine Phosphorylation Up-Regulation Virus Diseases Work Yam - dietary abstracting anergy genetic manipulation immune function improved in vivo mutant novel prevent programs promoter research study src-Family Kinases tool tumor

项目摘要

Program Director/Principal Investigator (Last, First, Middle): Gajewski, Thomas F. 1R01 AI080745-01A1 Revised Abstract Section Evidence suggests that one mechanism of peripheral immunologic tolerance occurs through the process of T cell anergy. Anergic I cells have been characterized to have a defect in TCR/CD28mediated Ras activation. Recent data have indicated that upregulated expression of lipid kinases of the diacylglycerol kinase (DGK) family, in particular DGK-u, participate in the suppression of (Ti j-<-0 L]' ADZ .Ti U,0 0(Q 0-0 COD c-0 'D- RasGRP-mediated Ras activation in the anergic state. Correlating with these changes is increased expression of the transcriptional regulator EGR2, and we have identified EGR-farnily binding sites in the putative DGK-ci promoter. The major goal of this proposal is to gain a detailed understanding of DGKs and EGR2 in controlling T cell activation and peripheral ._0 5a' coo u�r tow tolerance. In the first specific aim, the transcriptional regulation of the DGK-u will be elucidated, with a focus centered on EGR2. Novel tools for the genetic manipulation of primary T cells will be employed using the CAR Tg system. This includes transcriptional reporter adenoviral vectors, ... OUP adenoviruses for EGR2 expression, and a Cre adenovirus for conditional deletion of EGR2 in peripheral I cells. Supportive data will come from ChIP assays and gene expression profiling. In O-0 the second specific aim, the functional role of DGK-u tyrosine phosphorylabon will :E, �c' N-0 0-0 aux. be o03�-' targeted genes in the post-thymic I cell compartment. T cell activation properties, in vivo autoimmunity and anergy susceptibility, and possible improved tumor rejection in vivo will be addressed. In total, this work will characterize in detail a critical pathway in the control of peripheral tolerance, paving the way for development of pharmacologic agents to manipulate (gyp m='(Dm 3m03 a�fl` .., -'1 ascertained. Mutants of DGK-z will be analyzed functionally using adenoviral vectors and CAR Tg T cells in vitro, and transgenic mice expressing key DGK-u mutants in peripheral T cells will be studied for altered immune function in vivo. In the third specific aim, the consequence of elimination of EGR2 directly in peripheral T cells on peripheral tolerance will be investigated. These experiments will take advantage of the Cre-adenovirus approach to delete conditionally _=c� l17 immunologic tolerance in disease situations. 111 CDG'O V,12 oar coo COB

项目总监/首席调查员（最后一个，中间）：Gajewski，Thomas F. 1R01 AI080745-01A1 修订的摘要部分有证据表明，外周免疫耐受性的一种机制是通过T细胞消极过程发生的。 Anergic I细胞的表征是TCR/CD28介导的RAS激活中的缺陷。最近的数据表明，二酰基甘油激酶（DGK）家族的脂质激酶的表达更新，尤其是DGK-U，参与抑制（ti） J - <-0 l]'' adz .ti u，0 0（q 0-0 鳕鱼 C-0 'd- RASGRP介导的RAS激活在厌食状态。与这些变化相关的是转录调节剂EGR2的表达增加，我们已经确定了假定的DGK-CI启动子中的Egr-Farniely结合位点。该提案的主要目的是在控制T细胞激活和周围方面详细了解DGK和EGR2 ._0 5a' 公司 u 拖宽容。在第一个特定目的中，将阐明DGK-U的转录调节，重点集中于EGR2。使用CAR TG系统将使用用于原代T细胞遗传操作的新型工具。这包括转录记者腺病毒载体， ... OUP EGR2表达的腺病毒和外围I细胞中EGR2有条件缺失的CRE腺病毒。支持数据将来自CHIP分析和基因表达分析。在 O-0 第二个特定目的是DGK-U酪氨酸磷酸Labon的功能作用将：e， `c' N-0 0-0 辅助。是 o03.-' 胸膜后I细胞室中的靶向基因。 T细胞活化特性，体内自身免疫性和无抗敏感性以及可能改善体内肿瘤排斥的能力。总的来说，这项工作将详细描述一个控制外围耐受性的关键途径，为开发药物操纵的开发铺平了道路（GYP M ='（DM 3M03 Afl` ..，-'1 DGK-Z的突变体将在体外使用腺病毒载体和CAR TG T细胞进行功能分析，并且将研究表达关键DGK-U突变体在外周T细胞中的转基因小鼠，以在体内改变免疫功能。在第三个特定目的中，将研究直接在外周T细胞中消除EGR2对外周耐受性的结果。这些实验将利用CRE-腺病毒方法有条件删除 _ = c L17 疾病情况下的免疫耐受性。 111 cdg'o V，12 桨公司棒