Regulatory T cells in Autoimmune Disease

自身免疫性疾病中的调节性 T 细胞

• 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
• 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

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，我们提出了几种假设，这些假设为调节性T细胞的基本机制理解以及这些途径如何在自身免疫性疾病的动物模型和人类疾病I型糖尿病和多发性硬化症中改变提供了新的见解。此外，我们还开发了新的免疫方法，使用口服抗CD 3诱导调节性T细胞。该补助金将继续专注于这些基本目标，反映我们在过去五年中所取得的发现。在这次更新中，该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CD 4 + CD 25+亚群中的哪一个是缺陷的; 2)鉴定在“先天性”CD 4 + CD 25+调节性T细胞上表达的基因和蛋白质的“核心组”; 3)确定在糖尿病和MS患者血液和淋巴结中发现的调节性T细胞中哪些基因和蛋白质的“核心组”发生了改变; 4)了解口服抗CD 3和GRAIL转染T细胞的机制，以便在拨款的第五年将其转化为人类临床试验。其主要目标是开发特异性药物靶点和方法，以诱导自身免疫性疾病患者中有缺陷的调节性T细胞的功能。 概述：诸如I型糖尿病和多发性硬化症之类的疾病是复杂的遗传疾病，被认为是由针对发炎组织中的自身蛋白质的自身免疫反应引发的。为什么自身反应性T细胞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鞘仍然是一个主要问题。我们最近证明，在自身免疫性疾病患者的循环中，一种重要的“调节性”免疫细胞丢失。这项资助召集了一组研究人员，以了解为什么这些调节细胞功能失调，以及可以做些什么来恢复它们的功能。 项目1：CD 4 + CD 25 + T细胞的作用机制（FATHMAN，GARRISON C.） 描述（由申请人提供）： 这是一个竞争性的更新，作为一个U19为以前资助的U 01题为“CD 25+调节CD 4 + T细胞。“在为最初的U 01提供资金的情况下，实现了几个具体目标：最初（1）表达GRAIL（一个最近鉴定的无反应性基因），在静脉注射肽后，阻断IL-2转录并诱导无反应性，这是一种耐受形式，和（2）GRAIL表达提供了一种新的和有效的筛选小鼠无反应性表型的方法（和在人中），和（3）CD 4 + CD 25+抑制性T细胞参与这种形式的耐受性诱导，和第四（4）GVHD将被TcB的过继转移阻断，在支持的第二年增加。在这四个具体目标上发表了10篇文章，本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，将确定定义Treg核心转录组的“核心基因”。在该提议中，通过cDNA微阵列研究鉴定的这些“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，而“外周Treg亚群”暂时定义为在外周无反应性诱导条件下接触抗原的CD 4+抗原特异性T细胞，将按照以下四个具体目标的建议，进一步分析和研究： * 具体目标1：鉴定CD 4 + CD 25 + Treg基因的“核心集”，即Treg核心转录组。 * 具体目标2：外周诱导TcB的表征和核心转录组鉴定。 * 具体目标3：GRAIL转导物作为免疫治疗的TCL 4。 * 具体目的4：评价特定基因在移植物抗宿主病体内模型中对Treg功能的作用。 过继性细胞疗法的使用，特别是TGFAP的使用作为用于免疫调节的有用的潜在疗法，特别是在GVHD（过继性转移的自身免疫疾病的一种形式）的情况下，正迅速获得可信度。本项目中提出的研究将尝试鉴定定义THBG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模型来鉴定功能相关基因。这些研究所需的所有技术目前都在我们的实验室中实践。