High Throughput Identification of Treg Activating Molecules

Treg 激活分子的高通量鉴定

• 批准号: 8950523
• 负责人: Daniel J. Moore
• 金额: $ 23.23万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8950523
• 关键词: Adult; American; Animal Model; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Beta Cell; Biological; Biological Assay; Biological Markers; CD4 Positive T Lymphocytes; Cell physiology; Cells; Child; Clinic; Clinical Trials; Critical Pathways; Defect; Diagnosis; Disease; Equilibrium; FDA approved; Failure; Flow Cytometry; Glucose; Goals; Health; Human; Immune; Immune System Diseases; Immune system; Immunity; Inbred NOD Mice; Individual; Injury; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Lead; Lymphocyte; Mediating; Modeling; Mus; Organ; Outcome; Pathway interactions; Patients; Personal Satisfaction; Phosphotransferases; Positioning Attribute; Process; Regulation; Regulatory T-Lymphocyte; Research; Sampling; Self-Injurious Behavior; Societies; System; T-Cell Activation; T-Lymphocyte; Testing; Time; Tissues; Training; Translations; arm; base; cost; high throughput screening; human disease; in vivo; injured; islet; mouse model; novel; pre-clinical; prevent; research clinical testing; screening; small molecule libraries

DESCRIPTION (provided by applicant): Immune-mediated tissue injury is a primary mechanism of many disease processes that threaten human health. The immune system normally prevents self-injury through numerous control mechanisms, including a reliance on immune regulatory cells. Failure of these control processes can lead to autoimmunity. As a prototypical example of this autoimmune process, Type 1 diabetes (T1D) results from lymphocyte-mediated destruction of insulin-producing islet beta cells. In healthy individuals, the destructive power of the immune system is restrained by specific regulatory cells, but these cells fail to protect individuals with T1D. We propose that interventions to halt Type 1 diabetes and other immune disorders must train the immune system to control itself by restoring normal regulatory T cell function. Unfortunately, it has not been previously possible to rapidly screen candidate therapies to enhance regulatory T cell activation as there has been no biomarker that is unique to activated CD4 Tregs. Although there are markers of activated Tregs, these markers are also expressed by effector T cells; thus, once cells are activated, as is the case for many cells at all times during autoimmunity, it becomes impossible to determine the unique signature of activated Tregs, which confounds any disease-specific screening of Treg enhancing therapy. We have now identified a biomarker GARP that is uniquely expressed by activated CD4 Tregs, have demonstrated its insufficiency in T1D, and have combined this biomarker with other markers of Treg activation to develop a high throughput assay to identify lead compounds to enhance Treg activation. Applying this assay, we will now define new therapies and critical pathways to enhance Treg activation. Using high throughput flow cytometry, we will rapidly determine which compounds and pathways lead to Treg activation (Specific Aim 1). We will also begin translation of these approaches to the clinic by assessing induction of Treg activation in human PBMC's (Aim 1) and by demonstrating biological efficacy in animal models of autoimmune tissue injury and humanized systems (Specific Aim 2). These combined approaches give this application the potential for rapid impact on human diseases resulting from autoimmunity. Overall, our approach will lead to new understanding of the pathways involved in regulatory T cell activation and new pre-clinical approaches for correcting immune regulatory defects by accelerating our discovery of safe, individualized therapies that restore normal regulatory function to the immune system.

描述（由申请人提供）：免疫介导的组织损伤是威胁人类健康的许多疾病过程的主要机制。免疫系统通常通过许多控制机制来防止自我伤害，包括依赖免疫调节细胞。这些控制过程的失败可能导致自身免疫。作为这种自身免疫过程的典型例子，1型糖尿病（T1 D）是由淋巴细胞介导的对产生胰岛素的胰岛β细胞的破坏引起的。在健康个体中，免疫系统的破坏力受到特定调节细胞的抑制，但这些细胞无法保护T1 D患者。我们认为，阻止1型糖尿病和其他免疫疾病的干预措施必须通过恢复正常的调节性T细胞功能来训练免疫系统控制自身。不幸的是，以前不可能快速筛选候选疗法来增强调节性T细胞活化，因为没有活化的CD 4 Treg特有的生物标志物。尽管存在活化的TcR的标志物，但这些标志物也由效应T细胞表达;因此，一旦细胞被活化，如在自身免疫期间的所有时间对于许多细胞的情况，就不可能确定活化的TcR的独特特征，这混淆了Treg增强疗法的任何疾病特异性筛选。我们现在已经鉴定了由活化的CD 4 T细胞唯一表达的生物标志物GARP，已经证明了其在T1 D中的不足，并且已经将该生物标志物与Treg活化的其他标志物组合以开发高通量测定来鉴定增强Treg活化的先导化合物。应用这种检测方法，我们现在将定义新的疗法和关键途径来增强Treg激活。使用高通量流式细胞术，我们将快速确定哪些化合物和途径导致Treg激活（特异性目的1）。我们还将通过评估人PBMC中Treg活化的诱导（目标1）和通过在自身免疫组织损伤和人源化系统的动物模型中证明生物学功效（具体目标2）来开始将这些方法转化为临床。这些组合方法使该应用具有快速影响由自身免疫引起的人类疾病的潜力。总的来说，我们的方法将导致对调节性T细胞活化所涉及的途径的新理解，以及通过加速我们发现恢复免疫系统正常调节功能的安全，个性化疗法来纠正免疫调节缺陷的新临床前方法。