High Throughput Identification of Treg Activating Molecules

描述（由申请人提供）：免疫介导的组织损伤是许多威胁人类健康的疾病过程的主要机制。免疫系统通常通过多种控制机制来防止自我伤害，包括对免疫调节细胞的依赖。这些控制过程的失败可导致自身免疫。作为这种自身免疫过程的一个典型例子，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T1D患者。我们建议，干预1型糖尿病和其他免疫疾病必须通过恢复正常的调节性T细胞功能来训练免疫系统来控制自身。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，因为没有激活CD4 treg的独特生物标志物。虽然存在活化Tregs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的任何时候一样，就不可能确定活化Treg的独特特征，这使任何针对Treg增强治疗的疾病特异性筛选变得混乱。我们现在已经确定了一种生物标志物GARP，它是由活化的CD4 Treg唯一表达的，已经证明了它在T1D中的不足，并将这种生物标志物与其他Treg激活标志物结合起来，开发了一种高通量测定方法，以识别增强Treg激活的先导化合物。利用这种检测方法，我们现在将确定新的治疗方法和关键途径来增强Treg的激活。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（Specific Aim 1）。我们还将通过评估人类PBMC中Treg激活的诱导（目标1），以及通过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（特定目标2），开始将这些方法转化为临床应用。这些结合的方法使这一应用有可能对由自身免疫引起的人类疾病产生快速影响。总的来说，我们的方法将导致对调节性T细胞活化途径的新理解，并通过加速我们发现安全，个性化的治疗方法来恢复免疫系统的正常调节功能，从而纠正免疫调节缺陷的新临床前方法。

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.