Interrogating the ubiquitin pathway to understand and treat cytokine-induced beta-cell death in type 1 diabetes

探究泛素通路以了解和治疗 1 型糖尿病中细胞因子诱导的 β 细胞死亡

基本信息

批准号：
10668435
负责人：
Amit Choudhary
金额：
$ 54.3万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-30 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10668435
关键词：
Alberta province Apoptosis Area Autoimmune Diabetes Autoimmune Diseases Beta Cell Binding Biology Cell Death Cell Line Cell Survival Cell physiology Cellular biology Chemicals Clinical Complex Complications of Diabetes Mellitus Cytokine Signaling Development Diabetes Mellitus Diabetic mouse Disease Equilibrium Event Gene Expression Profiling Genetic Goals Heart Human Hypoglycemia Immune Immune Targeting In Vitro Individual Inflammatory Insulin Insulin-Dependent Diabetes Mellitus Interferon Type II Interleukin-1 beta JAK2 gene Kidney Label Lysine Measurable Measurement Mediating Methods Modeling Morbidity - disease rate Mus Mutate Non obese Outcome Pancreas Pathway interactions Patients Peripheral Nerves Phenotype Phosphorylation Phosphotransferases Post-Translational Protein Processing Process Proteins Proteomics Retina Risk Role STAT1 gene Signal Transduction Structure of beta Cell of islet TNF gene Therapeutic Tyrosine Ubiquitin Ubiquitination Universities activity-based protein profiling analog cellular targeting chemoproteomics cytokine early detection biomarkers improved in vivo innovation islet kinase inhibitor knock-down mortality risk mouse model novel therapeutic intervention preservation prevent response screening small molecule tool ubiquitin isopeptidase

项目摘要

PROJECT SUMMARY The loss of insulin-producing beta cells in the pancreas results in an absolute requirement for injected insulin, causing significant risks of mortality from hypoglycemia and morbidity from diabetic complications in peripheral nerves, the retina, the heart, and the kidney. A key goal of efforts to treat T1D is to stop this cellular attack, either by halting the immune mis-recognition of beta cells or by protecting beta cells from cell death. However, a critical barrier to progress in the field is a lack of complete understanding of the cellular events in the islet that contribute to the loss of beta-cell mass. Using a phenotypic screening approach, we discovered BRD0476, a compound that is selectively active against cytokine-mediated apoptosis. Further study of this compound revealed that it binds the deubiquitinase USP9X to halt JAK2 and STAT1 signaling in response to IFNγ. We determined that JAK2 can be rendered signaling incompetent by ubiquitination, and that by modulating USP9X, we can tip the balance toward reduced JAK2 kinase activity, even in the presence of IFNγ. These results point to an emerging role for ubiquitination in regulating beta-cell apoptosis in T1D, and suggest that a greater understanding of this process (and its potential dysregulation) in the early stages of T1D development could lead to 1) the ability to identify at-risk individuals, and 2) novel therapeutic strategies to preserve beta-cell mass in early-stage T1D. Using our probe BRD0476 and chemical biology tools not previously applied to islet biology, we will improve our understanding of the role of USP9X in beta-cell survival in vitro and in vivo through the following aims: In Aim 1, we will characterize the mode of JAK2 inhibition by USP9X in human islets. In Aim 2, we will assess effects of inhibiting USP9X-JAK2 (with BRD0476) on development and progression of autoimmune diabetes in a mouse model of type 1 diabetes. In Aim 3, we will profile deubiquitinase (DUB) expression and activity in human islets during early T1D development, using activity-based protein profiling (ABPP) and global ubiquitome measurements. The successful outcomes of this proposal are 1) a greater understanding of mechanisms to promote beta-cell survival in early T1D, and 2) a chemical probe to provide translational proof-of-concept. This project will set the stage for developing a biomarker of early-stage T1D development, as well as advanced therapeutic strategies for preventing beta-cell apoptosis in early-stage T1D, representing a potentially curative approach.

项目概要胰腺中产生胰岛素的β细胞的损失导致绝对需要注射胰岛素，导致外周低血糖死亡和糖尿病并发症发病的重大风险治疗 T1D 的一个关键目标是阻止这种细胞攻击。然而，通过阻止β细胞的免疫错误识别或保护β细胞免于细胞死亡。该领域取得进展的障碍是缺乏对胰岛细胞事件的完整了解，这些细胞事件有助于使用表型筛选方法，我们发现了 BRD0476，一种化合物。该化合物对细胞因子介导的细胞凋亡具有选择性活性。结合去泛素酶 USP9X 来停止响应 IFNγ 的 JAK2 和 STAT1 信号传导。 JAK2 可以通过泛素化而变得信号失灵，通过调节 USP9X，我们可以提示即使存在 IFNγ，这些结果也表明了一种新的趋势。泛素化在调节 T1D β 细胞凋亡中的作用，并表明对此有更深入的了解 T1D 发展早期阶段的过程（及其潜在的失调）可能导致 1) 识别高危个体，2) 在早期 T1D 中保留 β 细胞质量的新治疗策略。使用我们的探针 BRD0476 和以前未应用于胰岛生物学的化学生物学工具，我们将改进我们的通过以下目标了解 USP9X 在体外和体内 β 细胞存活中的作用：在目标 1 中，我们将描述 USP9X 在人类胰岛中抑制 JAK2 的模式。在目标 2 中，我们将评估 USP9X 的作用。抑制 USP9X-JAK2（与 BRD0476）对小鼠自身免疫性糖尿病的发生和进展在目标 3 中，我们将分析人类胰岛中的去泛素酶 (DUB) 表达和活性。在 T1D 早期发展过程中，使用基于活性的蛋白质分析 (ABPP) 和全局泛素组该提案的成功成果是 1) 更好地理解了机制。促进早期 T1D 中 β 细胞的存活，2) 提供翻译概念验证的化学探针。该项目将为开发早期 T1D 发展以及高级阶段的生物标志物奠定基础预防早期 T1D β 细胞凋亡的策略，代表了一种潜在的治愈方法方法。