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。免疫系统通常通过多种控制机制（包括依赖免疫调节细胞）来防止自伤。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 (T1D) 是由淋巴细胞介导的产生胰岛素的胰岛 β 细胞破坏所致。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 T1D 个体。我们建议，阻止 1 型糖尿病和其他免疫疾病的干预措施必须训练免疫系统通过恢复正常的调节性 T 细胞功能来控制自身。不幸的是，之前不可能快速筛选增强调节性 T 细胞激活的候选疗法，因为没有激活的 CD4 Tregs 特有的生物标志物。虽然存在激活Treg的标记，但这些标记也由效应T细胞表达；因此，一旦细胞被激活，就像许多细胞在自身免疫过程中一直存在的情况一样，就不可能确定激活的Treg的独特特征，这会混淆任何疾病特异性Treg增强疗法的筛选。我们现在已经鉴定出一种由激活的 CD4 Tregs 独特表达的生物标志物 GARP，并证明了其在 T1D 中的不足，并将该生物标志物与 Treg 激活的其他标志物结合起来，开发了一种高通量测定法来识别增强 Treg 激活的先导化合物。应用这种检测方法，我们现在将确定增强 Treg 激活的新疗法和关键途径。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（具体目标 1）。我们还将通过评估人类 PBMC 中 Treg 激活的诱导（目标 1）以及在自身免疫组织损伤和人源化系统的动物模型中展示生物功效（具体目标 2），开始将这些方法转化为临床。这些组合方法使该应用有可能对自身免疫引起的人类疾病产生快速影响。总体而言，我们的方法将通过加速我们发现恢复免疫系统正常调节功能的安全、个体化疗法，从而对调节性 T 细胞激活所涉及的途径产生新的理解，并带来纠正免疫调节缺陷的新临床前方法。