Dynamic analysis of beta cell attacks in response to virus infection in Human pancreatic slices

人胰腺切片中β细胞响应病毒感染的攻击的动态分析

• 批准号: 10674127
• 负责人: Mehdi Benkahla
• 金额: $ 27.45万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-27 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674127
• 关键词: 3-Dimensional; Antibodies; Autoimmune; Autoimmunity; Autologous; Autophagocytosis; Beta Cell; Blood; Capsid Proteins; Cell Death; Cell Survival; Cell physiology; Cells; Cellular Stress; Child; Coculture Techniques; Coxsackie Viruses; Coxsackievirus Infections; Data; Development; Diabetes Mellitus; Disease; Enterovirus; Environment; Environmental Risk Factor; Epidemiology; Etiology; Exocrine pancreas; Genetic Predisposition to Disease; Goals; Homeostasis; Human; Human Biology; Human Pathology; Immune; Immune mediated destruction; Immune system; In Vitro; Inbred NOD Mice; Incidence; Infection; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Link; Macrophage; Metabolic stress; Modeling; Morphology; Mus; Neurons; Pancreas; Patients; Peripheral Blood Mononuclear Cell; Physiology; Play; Predisposition; Reaction; Reporting; Research; Role; Signal Transduction; Slice; Structure of beta Cell of islet; T cell response; T-Cell Activation; T-Lymphocyte; Thick; Tissue Sample; Tissues; Viral; Virus; Virus Diseases; Virus Replication; autoimmune pathogenesis; autoreactive T cell; autoreactivity; blood glucose regulation; cell type; chronic autoimmune disease; diabetes pathogenesis; human tissue; immune activation; insulin dependent diabetes mellitus onset; insulin secretion; islet; islet cell antibody; meter; monocyte; non-diabetic; response; single-cell RNA sequencing; trend; viral RNA

PROJECT SUMMARY Type 1 diabetes (T1D) is a chronic autoimmune disease of unknown etiology leading to the destruction of pancreatic beta cells, which secrete insulin and regulate glucose homeostasis. The incidence of T1D has been increasing over the years, a trend inexplicable by only genetic predisposition, suggesting that environmental factors such as viruses play an important role in triggering the disease. Decades of research have provided epidemiological evidence for an association between T1D and viruses, particularly coxsackieviruses of the Enterovirus genus. Patients with T1D had higher rates of antibodies anti-CVB, viral RNA and viral capsid protein compared to non-diabetic. Studies have also shown that children who had developed islet autoantibodies had more CVB infections. Viral infection can impact beta cells through a direct cytolytic effect or by bystander activation of the immune system leading to gradual destruction. However, there is still little direct causal evidence from human pancreata that would establish a link between viral infections and T1D development. Although mouse studies have been very useful in understanding some of the mechanisms of the disease, there remain substantial gaps and differences to human pathology. Therefore, studies on Human pancreatic tissue samples become essential. Pancreas tissue slices are 150-200 µm thick sections of living human donor pancreatic tissue. They enable functional analysis of intact islets in their endogenous environment and 3D morphological assessment of the endocrine and exocrine pancreas as well as the blood vasculature, neurons and tissue resident immune cells. In preliminary studies, we have been able to culture and infect Human pancreatic slices using coxsackievirus B3 (CVB3)-GFP and identify the beta cells and tissue-resident macrophages as targets. Thus, we hypothesize that virus infections induce changes in tissue-resident macrophages which then induce T cells to attack and destroy beta cells. Aim 1 will evaluate the impact of CVB3-GFP infection on beta cells function and viability. Aim 2 will characterize tissue-resident macrophages during CVB3 infection by single cell RNA-seq. Aim 3 will examinate if the infection could lead to an immune destruction of beta cells when cocultured with HLA matched or autologous PBMCs. Aim 4 will characterize the impact of CVB3 infection on autophagy in infected Human pancreatic slices. Overall, this effort will generate a better understanding of the consequences of viral infections in the pancreas and its role in the onset of T1D.

项目摘要 1型糖尿病（T1 D）是一种病因不明的慢性自身免疫性疾病，导致糖尿病患者体内 胰腺β细胞，其分泌胰岛素并调节葡萄糖稳态。T1 D的发病率一直是 多年来不断增加，这一趋势只能通过遗传易感性来解释，这表明环境 病毒等因素在引发该病方面起着重要作用。几十年的研究提供了 流行病学证据表明，T1 D与病毒，特别是柯萨奇病毒之间存在关联。 肠道病毒属。T1 D患者抗CVB抗体、病毒RNA和病毒衣壳蛋白的检出率较高 与非糖尿病患者相比。研究还表明，患有胰岛自身抗体的儿童， 更多CVB感染病毒感染可通过直接溶细胞效应或旁观者作用影响β细胞 免疫系统的激活导致逐渐的破坏。然而，直接的因果证据仍然很少 这将建立病毒感染和T1 D发展之间的联系。虽然 小鼠研究在理解疾病的某些机制方面非常有用，但仍然存在 与人类病理学的巨大差距和差异。因此，对人胰腺组织样本的研究 变得必不可少。胰腺组织切片是150-200 µm厚的活体人类供体胰腺组织切片。 它们能够在其内源性环境和3D形态学中对完整胰岛进行功能分析 内分泌和外分泌胰腺以及血管系统、神经元和组织的评估 常驻免疫细胞在初步研究中，我们已经能够培养和感染人胰腺切片， 使用柯萨奇病毒B3（CVB 3）-GFP，并鉴定β细胞和组织驻留巨噬细胞作为靶。 因此，我们假设病毒感染诱导组织驻留巨噬细胞的变化，然后诱导T 攻击并摧毁β细胞。目的1将评估CVB 3-GFP感染对β细胞的影响 功能和可行性。目的2将通过单个细胞来表征CVB 3感染期间组织驻留的巨噬细胞 RNA测序目标3将检查当共培养时，感染是否会导致β细胞的免疫破坏 与HLA匹配或自体PBMC。目的4将描述CVB 3感染对自噬的影响 在受感染的人类胰腺切片中总的来说，这一努力将使人们更好地了解其后果 胰腺中的病毒感染及其在T1 D发病中的作用。