Regulatory T cells in Autoimmune Disease

自身免疫性疾病中的调节性 T 细胞

基本信息

批准号：
8136146
负责人：
CHARLES GARRISON FATHMAN
金额：
$ 49.08万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8136146
关键词：
Address Adoptive Transfer Animal Model Antigens Autoimmune Diseases Autoimmune Responses Autoimmunity Biological Assay Blood Blood Circulation CD4 Antigens CD4 Positive T Lymphocytes Cells Clinical Trials Complex Development Diabetes Mellitus Disease Drug Delivery Systems Funding Gene Expression Profile Gene Proteins Gene Silencing Genes Genetic Transcription Goals Grant Hereditary Disease Human IL2RA gene Immune Immunotherapeutic agent Immunotherapy Insulin Insulin-Dependent Diabetes Mellitus Interleukin-2 Islet Cell Methods Modeling Multiple Sclerosis Mus Myelin Oral Pathway interactions Patients Peptides Peripheral Phenotype Prevention Proteins Publishing RNA Interference Regulatory T-Lymphocyte Research Personnel Role Suppressor-Effector T-Lymphocytes T-Cell Proliferation T-Lymphocyte Techniques Technology Tissues Transfection Translations anergy autoreactive T cell cDNA Arrays functional genomics graft vs host disease human disease immunoregulation in vitro Assay in vitro testing in vivo in vivo Model insight knock-down loss of function lymph nodes mRNA Expression man novel

项目摘要

DESCRIPTION (provided by applicant): Under funding for the original U19, we addressed several hypotheses that have provided new insights into the fundamental, mechanistic understanding of regulatory T cells and how these pathways are altered in animal models of autoimmune disease and in the human diseases type I diabetes and multiple sclerosis. In addition, we have developed novel immunotherapeutic approaches to induce regulatory T cells using oral anti-CD3. The grant will continue to focus on these fundamental aims, reflecting the discoveries we have made over the past five years. In this renewal, the overall goals of this Autoimmunity Prevention Center Project are: 1) To determine, in human autoimmune disease, which of the CD4+CD25+ subsets of DR+ and DR" regulatory T cells are defective; 2) The identification of a "core set" of genes and proteins that are expressed on the "innate", CD4+CD25+ regulatory T cells; 3) To determine which of the "core sets" of genes and proteins are altered in the regulatory T cells found in blood and lymph nodes in patients with diabetes and MS; 4) To understand the mechanism of oral anti-CD3 and GRAIL transfection in T cells that allows the translation into human clinical trials by year five of the grant. The key goal is the development of specific drug targets and methods to induce the function of defective regulatory T cells in patients with autoimmune disease. Lay Summary: Diseases such as type I diabetes and multiple sclerosis are complex genetic diseases thought to be initiated by an autoimmune response directed against self-proteins in the inflamed tissue. Why autoreactive T cells attack the insulin producing islet cells in diabetes or the myelin in multiple sclerosis remains a major question. We recently demonstrated there is a loss of an important "regulatory" immune cell in the circulation of patients with autoimmune disease. This grant assembles a group of investigators to understand why these regulatory cells are dysfunctional, and what can be done to restore their function. PROJECT 1: Mechanism of action of CD4+CD25+ T cells (FATHMAN, GARRISON C.) DESCRIPTION (provided by applicant): This is a competing renewal as a U19 for a previously funded U01 entitled, "CD25+ Regulator CD4+ T Cells." Under funding for the original U01, several specific aims were addressed: initially (1) that expression of GRAIL (a recently identified anergy gene), following peptide administration iv, blocks IL-2 transcription and induces anergy, a form of tolerance, and (2) that GRAIL expression provides a novel and effective screen for the anergic phenotype in mice (and in man), and (3) that CD4+CD25+ suppressor T cells were involved in this form of tolerance induction, and a fourth (4) GVHD will be blocked by adoptive transfer of Tregs, was added in the second year of support. Ten articles were published on these four specific aims and this proposal will extend these studies as follows: Using microarray and RNAi technology for mRNA expression and gene silencing, the "core genes" that define Treg core transcriptome will be identified. In this proposal, these "core Treg" genes, identified by cDNA microarray studies, will be validated by functional genomics (RNAi) and tested in vitro in T cell proliferation assays and in vivo in a model of GVHD, and the "peripheral Treg subset," provisionally defined as CD4+ antigen specific T cells that contact antigen under anergy inducing conditions in the periphery, will be further characterized and studied as proposed in the following four specific aims: * Specific Aim 1: Identification of a "core set" of CD4+CD25+ Treg genes, the Treg core transcriptome. * Specific Aim 2: Characterization of peripherally induced Tregs and core transcriptome identification. * Specific Aim 3: GRAIL transductants as Tregs for immunotherapy. * Specific Aim 4: To evaluate the role of specific genes on Treg function in an in vivo model of graft vs. host disease. The use of adoptive cellular therapy, in particular use of Tregs is rapidly gaining credibility as a useful potential therapy for immunoregulation, in particular in the setting of GVHD, a form of adoptively transferred autoimmune disease. Studies proposed in this project will attempt to identify the "core" set of genes that define Tregs and then using RNA silencing techniques, attempt to identify functionally relevant genes through knock down and loss of function assays in vitro and using a model of GVHD in vivo. All of the techniques required for these studies are currently practiced in our labs.

描述（由申请人提供）：在原始 U19 的资助下，我们提出了几个假设，这些假设为对调节性 T 细胞的基本机制理解以及这些途径在自身免疫性疾病动物模型以及人类疾病 I 型糖尿病和多发性硬化症中如何改变提供了新的见解。此外，我们还开发了新的免疫治疗方法，使用口服抗 CD3 来诱导调节性 T 细胞。这笔赠款将继续关注这些基本目标，反映我们在过去五年中取得的发现。在本次更新中，自身免疫预防中心项目的总体目标是： 1) 确定在人类自身免疫性疾病中，DR+和DR”调节性T细胞的CD4+CD25+亚群中哪些存在缺陷； 2) 鉴定在“先天”CD4+CD25+调节性T细胞上表达的基因和蛋白质的“核心组”； 3) 确定糖尿病和多发性硬化症患者血液和淋巴结中的调节性 T 细胞中哪些基因和蛋白质的“核心组”发生了改变； 4) 了解 T 细胞中口服抗 CD3 和 GRAIL 转染的机制，以便在资助的第五年转化为人体临床试验。主要目标是开发特定的药物靶点和方法来诱导自身免疫性疾病患者有缺陷的调节性 T 细胞的功能。简单总结：I 型糖尿病和多发性硬化症等疾病是复杂的遗传性疾病，被认为是由针对发炎组织中自身蛋白的自身免疫反应引发的。为什么自身反应性 T 细胞会攻击糖尿病中产生胰岛素的胰岛细胞或多发性硬化症中的髓磷脂仍然是一个主要问题。我们最近证明，自身免疫性疾病患者的循环中存在一种重要的“调节”免疫细胞的丢失。这笔赠款聚集了一组研究人员，以了解这些调节细胞功能失调的原因，以及可以采取哪些措施来恢复其功能。项目 1：CD4+CD25+ T 细胞的作用机制（FATHMAN、GARRISON C.）描述（由申请人提供）：这是对先前资助的 U01 的 U19 的竞争性更新，题为“CD25+ 调节器 CD4+ T 细胞”。在原始 U01 的资助下，解决了几个具体目标：最初 (1) GRAIL（一种最近发现的无反应基因）的表达，在静脉注射肽后，阻断 IL-2 转录并诱导无反应（一种耐受形式），以及（ 2) GRAIL 表达为小鼠（和人类）的无反应表型提供了一种新颖且有效的筛选方法，(3) CD4+CD25+ 抑制性 T 细胞参与了这种形式的耐受诱导，第四个(4) GVHD将通过Tregs的过继转移来阻止，在支持的第二年添加。关于这四个具体目标已发表了 10 篇文章，该提案将扩展这些研究如下：使用微阵列和 RNAi 技术进行 mRNA 表达和基因沉默，将鉴定定义 Treg 核心转录组的“核心基因”。在该提案中，这些通过 cDNA 微阵列研究鉴定的“核心 Treg”基因将通过功能基因组学 (RNAi) 进行验证，并在 T 细胞增殖测定中进行体外测试，并在 GVHD 模型中进行体内测试，以及“外周 Treg 子集” “，”暂时定义为在外周无反应诱导条件下接触抗原的 CD4+ 抗原特异性 T 细胞，将按照以下四个具体目标进一步表征和研究： * 具体目标 1：鉴定 CD4+CD25+ Treg 基因的“核心集”，即 Treg 核心转录组。 * 具体目标 2：外周诱导性 Tregs 的表征和核心转录组鉴定。 * 具体目标 3：GRAIL 转导子作为免疫治疗的 Tregs。 * 具体目标 4：在移植物抗宿主疾病的体内模型中评估特定基因对 Treg 功能的作用。过继性细胞疗法的使用，特别是 Tregs 的使用，作为一种有用的潜在免疫调节疗法，正在迅速获得可信度，特别是在 GVHD（一种过继转移的自身免疫性疾病）的情况下。该项目提出的研究将尝试识别定义 Tregs 的“核心”基因组，然后使用 RNA 沉默技术，尝试通过体外敲低和功能丧失测定以及使用体内 GVHD 模型来识别功能相关基因。这些研究所需的所有技术目前都在我们的实验室中实践。