Abnormal Extracellular Vesicles and Particles from Human Islets Impact T1D progression

来自人类胰岛的异常细胞外囊泡和颗粒影响 T1D 进展

• 批准号: 10754074
• 负责人: Shuibing Chen
• 金额: $ 72.91万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-21 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10754074
• 关键词: Alpha Cell; Antigen Presentation; Antigen-Presenting Cells; Apoptotic; Beta Cell; Binding; Biological Markers; Biology; Blood Vessels; Body Fluids; Cell Communication; Cell Line; Cell Survival; Cell membrane; Cell physiology; Cells; Coxsackie Viruses; DNA; Databases; Development; Diagnosis; Endocrine; Endothelial Cells; Field Flow Fractionation; Functional disorder; Heterogeneity; Human; Immune; Insulin-Dependent Diabetes Mellitus; Islet Cell; Lipid Bilayers; Lipids; MLLT2 gene; Macrophage; Mediating; Mediator; Membrane; Membrane Proteins; Methods; MicroRNAs; Microfluidic Microchips; Molecular; Molecular Profiling; Organ; Organoids; Outcome; Pancreas; Pathogenicity; Pathologic; Patient Care; Patients; Plasma; Proteins; Proteomics; RNA; Role; Sampling; Slice; Stress; Structure; Technology; Testing; Tissues; Ultracentrifugation; Vascular blood supply; Vascularization; autoreactivity; cell type; cytokine; diabetes pathogenesis; exosome; extracellular; extracellular vesicles; gain of function; human pluripotent stem cell; insulin dependent diabetes mellitus onset; islet; mouse model; nanoparticle; novel marker; novel therapeutic intervention; novel therapeutics; particle; prevent; senescence; signature molecule; superresolution imaging

Abstract. Extracellular vesicles and particles (EVPs) are a group of heterogeneous nanoparticles that are secreted into the extracellular milieu by many types of cells. EVPs have emerged as important mediators in cell-to-cell communications within organs. Recent studies have suggested that EVPs might be involved in β cell-immune cell interaction and β cell dysfunction and survival in type 1 diabetes (T1D). However, many of these studies were performed using mouse models and not focused on human samples. Hence, there is a unmet need to understand the contribution of human islet/pancreas-associated EVPs in T1D progression. Here, we have assembled an interdisciplinary team, including experts in islet biology (Chen, Liu), EVP biology (Lyden), vascular biology (Rafii), and T1D patient care (Antal), to test the hypothesis that abnormal EVPs of pathogenic human islets and pancreatic tissues disrupt human islet function and survival and trigger T1D progression. We have performed extensive proteomics analyses of EVPs derived from human β cell lines cultured in T1D conditions. In addition, by comparing the plasma membrane protein profiles of human β cell line and primary human islet-derived EVPs with an EVP proteomic database containing 426 human samples, we have identified several plasma membrane protein candidates to isolate β cell-specific EVPs. Here, we will use multiple platforms, including primary human islets from healthy, pre-T1D, and T1D donors, human pluripotent stem cell (hPSC)-derived islet organoids, perfusable microfluidic devices containing vascularized islets, and pancreatic slice culture to systematically examine the role of human islet/pancreas-associated EVPs in islet cell function and survival during T1D progression. In this proposal, we expect to identify membrane-bound EVP protein panels to purify EVPs from human α, β cells, islets, and pancreatic tissue. We will define the EVP protein and RNA profiles and identify signature molecules associated with islet status during T1D progression. Moreover, we will systemically dissect the roles of human islet/pancreas EVPs in human β cell function and survival, as well as their role in the crosstalk between β cells and peri-islet cell niche. Overall, this study will facilitate the identification of novel “biomarkers” for T1D diagnosis and guide the development of new therapeutic strategies to treat and prevent the progression of T1D.

抽象。 细胞外囊泡和颗粒（EVP）是一组分泌到细胞内的异质纳米颗粒。 许多类型的细胞的细胞外环境。EVP已经成为细胞间相互作用的重要介质。 机关内部的沟通。最近的研究表明，EVP可能参与β细胞免疫 1型糖尿病（T1 D）中的细胞相互作用和β细胞功能障碍和生存。然而，许多研究 使用小鼠模型进行，而不是集中在人类样本上。因此，有一个未满足的需要， 了解人类胰岛/胰腺相关EVP在T1 D进展中的作用。 在这里，我们聚集了一个跨学科的团队，包括胰岛生物学专家（陈，刘），EVP生物学 （Lyden）、血管生物学（Rafii）和T1 D患者护理（Antal），以检验以下假设： 致病性人胰岛和胰腺组织破坏人胰岛功能和存活， T1 D进展。我们对来源于人β细胞系的EVP进行了广泛的蛋白质组学分析 在T1 D条件下培养。此外，通过比较人β细胞系的质膜蛋白质谱， 和原代人胰岛衍生的EVP与包含426个人样本的EVP蛋白质组学数据库，我们 已经鉴定了几种质膜蛋白候选物以分离β细胞特异性EVP。在这里，我们将使用 多个平台，包括来自健康、T1 D前和T1 D供体的原代人胰岛，人多能 干细胞（hPSC）衍生的胰岛类器官，含有血管化胰岛的可灌注微流体装置，以及 胰腺切片培养，系统检测人胰岛/胰腺相关EVP在胰岛细胞中的作用 T1 D进展期间的功能和存活率。 在本研究中，我们期望鉴定膜结合的EVP蛋白组，以从人α，β 细胞、胰岛和胰腺组织。我们将确定EVP蛋白和RNA谱，并确定签名 与T1 D进展期间胰岛状态相关的分子。此外，我们将系统地剖析角色 人胰岛/胰腺EVP在人β细胞功能和存活中的作用，以及它们在人胰岛/胰腺EVP之间的串扰中的作用。 β细胞和胰岛周围细胞龛。总体而言，本研究将有助于识别T1 D的新型“生物标志物” 诊断和指导新的治疗策略的开发，以治疗和预防T1 D的进展。