The Effects of Low-Dose IL-2 Therapy on Beta Cell Dysfunction in Type 1 Diabetes

低剂量 IL-2 治疗对 1 型糖尿病 β 细胞功能障碍的影响

• 批准号: 10612366
• 负责人: Farhan Qureshi
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612366
• 关键词: Address; Affect; Aftercare; American; Apoptosis; Autoimmune Diseases; Autoimmunity; Beta Cell; Biology; Biomedical Research; Blood; Blood Glucose; Cell Communication; Cell Count; Cell Death; Cell physiology; Cells; Chimeric Proteins; Clinical; Clinical Research; Clinical Trials; Cyclosporine; Data; Diabetes Mellitus; Diagnosis; Disease; Disease Progression; Dose; Endocrine; Endoplasmic Reticulum; Environment; Fasting; Fluorescence-Activated Cell Sorting; Foundations; Functional disorder; Goals; Half-Life; Health; Homeostasis; IL2RA gene; Immune; Immune Tolerance; Immunosuppressive Agents; Immunotherapy; Impairment; In Situ; In Vitro; Inbred NOD Mice; Individual; Infiltration; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Interleukin-2; Intervention; Investments; Islets of Langerhans; Knowledge; Lymphocyte; Mediating; Modeling; Monitor; NOR Mouse; Non obese; Pancreas; Pathogenesis; Patients; Pattern; Phase I/II Clinical Trial; Population; Pre-Clinical Model; Preparation; Prevention; Proliferating; Recombinant Interleukin-2; Regulatory T-Lymphocyte; Science; Signal Transduction; Slice; Stains; Strategic Planning; Stress; Testing; Time; Tissues; Treatment Efficacy; United States National Institutes of Health; Work; analog; autoimmune pathogenesis; bench to bedside; biological adaptation to stress; blood glucose regulation; cytokine; diabetes pathogenesis; diabetic; effector T cell; efficacy trial; functional restoration; glucose tolerance; immune cell infiltrate; immunoregulation; insulin secretion; insulitis; islet; neglect; novel; novel strategies; novel therapeutics; preclinical study; preclinical trial; preservation; prevent; response; restoration; single-cell RNA sequencing

Project Summary Type 1 diabetes (T1D) is a devastating autoimmune disease affecting over 1.25 million Americans that currently has no cure. Therapies which target autoimmunity in T1D have shifted away from broadly immunosuppressive agents such as cyclosporine, and now focus on enhancing immune tolerance. Low-doses of interleukin-2 (IL-2) have been shown in pre-clinical and clinical trials to induce proliferation of regulatory Tcells (Tregs) which promote immune tolerance and prevent autoimmune destruction of the insulin producing beta cell. Recently, it has been shown in preclinical models that an IL-2/CD25 fusion protein induces greater proliferation of Tregs and has a longer half-life than traditional recombinant IL-2. Studies on IL-2 and its analogues have focused on mechanisms of immune tolerance and how it prevents loss of beta cell mass. Recent clinical findings demonstrate that insulin secretion is impaired at least five years before diagnosis, while beta cell mass is still preserved, suggesting that beta cell dysfunction, in addition to loss of beta cell mass, is critically important to the pathogenesis of T1D. It is currently unknown how IL-2/CD25 therapy affects beta cell dysfunction. This represents a critical gap in knowledge that must be addressed. Preliminary data shows that IL-2/CD25 induces changes in the local immune infiltrate, increasing the proportion of Tregs and decreasing the proportion of effector T cells (Teffs) at the islets. In vitro studies have shown that pro-inflammatory cytokines are released by infiltrating Teffs and cause beta cell dysfunction via stress on the endoplasmic reticulum (ER). Therefore, I hypothesize that IL-2/CD25 works to delay T1D progression by preventing cytokine-mediated beta cell dysfunction, not just by averting cell death. I plan to test this hypothesis in the following specific aims: 1) Determine how IL-2/CD25 treatment preserves islet function and glucose homeostasis, 2) Determine how IL- 2/CD25 treatment preserves islet tissue homeostasis. Under the first aim, I will use a novel approach using living pancreas slices to analyze changes in insulin secretion and intracellular Ca2+ dynamics after IL-2/CD25 treatment. Under the second aim, I will identify the specific mechanisms involved in restoring islet homeostasis after IL-2/CD25 therapy by using immunostaining and single-cell RNA sequencing for markers of stress, proliferation, and dedifferentiation. My proposed study is significant because it will yield new information about how changes in local immune infiltration mediated by IL-2/CD25 affect islet biology. This contribution is significant because it will provide fundamental knowledge that will complete and revise models about immunotherapies for T1D. This study supports the strategic plans of the National Institutes of Health which include advancing opportunities in biomedical research by investing in fundamental science and developing treatments/cures for disease.

项目摘要 1型糖尿病（T1 D）是一种毁灭性的自身免疫性疾病，目前影响超过125万美国人， 无法治愈靶向T1 D自身免疫的治疗已经从广泛的免疫抑制转向 环孢菌素等药物，现在专注于增强免疫耐受性。低剂量白细胞介素-2（IL-2） 已经在临床前和临床试验中显示诱导调节性T细胞（Tcells）增殖， 促进免疫耐受并防止产生胰岛素的β细胞的自身免疫破坏。近日 已经在临床前模型中显示，IL-2/CD 25融合蛋白诱导TcB更大的增殖， 具有比传统重组IL-2更长的半衰期。IL-2及其类似物的研究主要集中在 免疫耐受机制以及它如何防止β细胞质量损失。近期临床发现 表明胰岛素分泌受损至少五年前诊断，而β细胞质量仍然是 保存，表明β细胞功能障碍，除了β细胞质量的损失，是至关重要的， T1 D的发病机制目前尚不清楚IL-2/CD 25治疗如何影响β细胞功能障碍。这 这是一个必须解决的知识缺口。初步数据显示IL-2/CD 25诱导 局部免疫浸润的变化，增加了T细胞的比例，减少了T细胞的比例。 效应T细胞（Tefs）在胰岛。体外研究表明，促炎细胞因子是由 在一些实施方案中，Teff可通过对内质网（ER）的应激而浸润Teff并引起β细胞功能障碍。所以我 假设IL-2/CD 25通过阻止精氨酸介导β细胞 而不仅仅是避免细胞死亡。我计划在以下具体目标中检验这一假设：1） 确定IL-2/CD 25治疗如何保护胰岛功能和葡萄糖稳态，2）确定IL-2/CD 25治疗如何保护胰岛功能和葡萄糖稳态， 2/CD 25治疗保持胰岛组织稳态。在第一个目标下，我将使用一种新颖的方法， 胰腺切片分析IL-2/CD 25后胰岛素分泌和细胞内Ca 2+动力学的变化 治疗在第二个目标下，我将确定恢复胰岛稳态的具体机制 在通过使用免疫染色和单细胞RNA测序的应激标志物进行IL-2/CD 25治疗后， 增殖和去分化。我提出的研究是重要的，因为它将产生新的信息 IL-2/CD 25介导的局部免疫浸润的变化如何影响胰岛生物学。这一贡献意义重大 因为它将提供基础知识，完善和修订免疫疗法的模型， T1 D。这项研究支持美国国立卫生研究院的战略计划，其中包括推进 通过投资基础科学和开发治疗方法，为生物医学研究提供机会。 疾病