Implications of Changes in Islet Exosomal Cargo in Type 1 Diabetes

1 型糖尿病中胰岛外泌体货物变化的影响

• 批准号: 10708900
• 负责人: Decio laks Eizirik
• 金额: $ 68.43万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708900
• 关键词: Antibody Therapy; Antigens; Apoptosis; Autoimmune; Autoimmune Diabetes; Autoimmunity; Beta Cell; Binding; Biogenesis; Biological Markers; Biology; C-Peptide; CD8-Positive T-Lymphocytes; Cell Communication; Cell Differentiation process; Cell Line; Cell Survival; Cell physiology; Cells; Cellular Stress; Cessation of life; Chemicals; Clinical; Cytotoxic T-Lymphocytes; Data; Development; Disease; Etiology; Exocytosis; Goals; Health; Heterogeneity; Housing; Human; Immune; Immune Evasion; Immune checkpoint inhibitor; Immune mediated destruction; In Vitro; Inbred NOD Mice; Incidence; Individual; Inflammatory; Insulin-Dependent Diabetes Mellitus; Interferons; Intervention; Intravenous; Knowledge; Link; Lymphocyte; Mediator; Membrane; Messenger RNA; Microfluidic Microchips; Molecular; Multivesicular Body; Pancreas; Parents; Pathway interactions; Persons; Physiological; Plasma; Play; Pre-Clinical Model; Proteins; Regulation; Research; Residual state; Risk; Signal Pathway; Signal Transduction; Slice; Specificity; Sphingomyelinase; Stimulus; Stress; Surface; T-Cell Activation; T-Lymphocyte; Testing; Work; autoimmune pathogenesis; clinical biomarkers; cytokine; density; diabetogenic; diagnostic strategy; exosome; extracellular vesicles; genetic manipulation; immune checkpoint; in vivo; induced pluripotent stem cell; insulin dependent diabetes mellitus onset; islet; islet cell antibody; nanoparticle; neoplastic cell; non-diabetic; novel; novel diagnostics; novel marker; overexpression; paracrine; preservation; prevent; programmed cell death ligand 1; programmed cell death protein 1; receptor; response; stressor; targeted treatment; therapeutic target; transcription factor; uptake

PROJECT SUMMARY Extracellular vesicles (EVs) are membrane bound nanoparticles that can interact with other cells as a means of cell:cell communication. Emerging data suggest that β cell-derived exosomes, an EV subtype released by exocytosis of multivesicular bodies, may act as paracrine effectors in islet health. Given the potential for exosomes to play a role in β cell communication with surrounding cells in the islet microenvironment, a critical need exists for deliberate research defining mechanisms of β cell exosome biology under physiologic and pathophysiologic circumstances. β cell expression of the transmembrane immune checkpoint protein death-ligand 1 (PD-L1) plays a key role to support β cell survival in type 1 diabetes (T1D). Preliminary data suggest that β cell exosome membranes carry PD-L1, and that exosomal PD-L1 is upregulated by cytokine treatment of parent β cells. Our central hypothesis is that stressed β cells in the T1D microenvironment alter their exosomal contents to include protective mechanisms aimed at evading β cell destruction associated with autoimmunity. Aim 1 will elucidate the mechanistic etiology of altered β cell exosome PD-L1 cargo. We hypothesize that proinflammatory interferon signaling activates molecular regulators of intracellular β cell PD-L1 that, in concert with mediators of exosome biogenesis, increase total exosomal PD-L1. Chemical and genetic manipulation will be used to test impacts of these pathways on β cell exosomal PD-L1. Aim 2 will test the hypothesis that via transfer to or interaction with surrounding β cells and T cells, β cell exosomal PD-L1 loading is a protective mechanism shielding β cells from autoimmune destruction. Use of β cells differentiated from a human induced pluripotent stem cell line overexpressing GFP-tagged PD-L1 will allow for direct tracking of PD-L1 transfer and binding to other β cells and CD8+ T cells. Aim 3 will employ a novel microfluidic device to test the hypothesis that PD-L1+ exosome release will be increased in humans with or at risk for T1D. We will quantify differences in total and islet-derived EV PD-L1 in human plasma or pancreas slice media and compare to nondiabetic controls. Testing will also be performed in plasma from individuals with longstanding T1D with or without residual detectable C-peptide to determine if plasma islet-derived EV PD-L1 is linked to functional β cell survival. This work will lead to a paradigm shift in the field’s understanding of β cell communication with surrounding cells, and determine the clinical potential of islet-derived PD-L1 exosome cargo as a therapeutic targeting β cell survival or a biomarker to dissect T1D disease-related heterogeneity.

项目总结 胞外囊泡(EV)是一种膜结合的纳米颗粒，可以与其他细胞相互作用 作为细胞的一种手段：细胞通讯。新出现的数据表明，β细胞衍生的外切体，一种 EV亚型通过多囊泡体的胞吐作用而释放，可能作为旁分泌效应器 小岛健康。鉴于外切体可能在β细胞与 胰岛微环境中的细胞周围存在刻意研究的迫切需要 生理和病理生理条件下β细胞外切体生物学机制的研究 情况。跨膜免疫检查点蛋白死亡配体在β细胞中的表达 1(PD-L1)在支持1型糖尿病(T1D)β细胞存活中起关键作用。初步数据 提示β细胞胞外体膜携带PD-L1，胞外体PD-L1上调 通过细胞因子处理亲本β细胞。我们的中心假设是T1D中的应激β细胞 微环境改变它们的胞外体内容物以包括针对 逃避与自身免疫相关的β细胞破坏。目标1将阐明 β细胞外切体PD-L1货代改变的病因学。我们假设促炎干扰素 信号激活细胞内β细胞PD-L1的分子调节器，与 外切体生物发生的介体，增加总外切体PD-L1。化学和遗传 操作将被用来测试这些通路对β细胞外体PD-L1的影响。目标2 将检验这样一种假设，即通过转移到周围的β细胞和T细胞或与其相互作用，β 细胞外体PD-L1负载是保护β细胞免受自身免疫的一种保护机制 毁灭。人诱导多能干细胞系分化出的β细胞的应用 过表达GFP标记的PD-L1将允许直接跟踪PD-L1的转移和结合 其他β细胞和CD8+T细胞。AIM 3将使用一种新的微流控设备来测试 假设患有T1D或有T1D风险的人PD-L1+外切体释放增加。 我们将量化人类血浆或胰腺中总的和胰岛来源的EVPD-L1的差异 切片，并与非糖尿病对照组进行比较。测试也将在血浆中进行， 长期存在T1D的个体是否存在残留的可检测到的C肽以确定 血浆胰岛衍生EV PD-L1与功能性β细胞存活有关。这项工作将导致 该领域对β细胞与周围细胞交流的理解发生了范式转变 确定胰岛来源的PD-L1外切小体作为治疗靶点的临床潜力 β细胞存活或作为解剖T1D疾病相关异质性的生物标记物。