Interrogation of dynamic RNA modifications in beta cells in type 1 diabetes

项目总结 全球糖尿病患者的数量预计将从2015年的4.15亿增加到6.42亿 在2040年。治疗这种疾病的医疗保健费用占全球卫生支出的12%， 这仍然是一个巨大的经济负担(以色列国防军世界地图集，2015年)。1型糖尿病(T1 DM)构成 疾病负担的10%-15%，是一种自身免疫性疾病，被认为是由遗传或 儿童早期的环境因素，导致抗体介导的胰岛素产生的破坏 胰腺β细胞--导致危及生命的高血糖。相比之下，2型糖尿病(T2 DM) 在生命的后期发展，是由于靶组织中的胰岛素抵抗和炎症与 β细胞的补偿不足。 触发最终导致β细胞死亡的一连串事件的确切机制仍然是 没有完全理解。一些观察结果再次激发了人们对β细胞本身作为靶子的兴趣 可能引发疾病进程的组织。在这方面，最近发现的RNA甲基化是一种 潜在的调节机制，有助于细胞适应细胞内的快速变化 环境提供了一种挑衅性的方法来研究先于 1型糖尿病表型的发展。我们试图询问动态甲基化的变化 RNA，特别是β细胞中的N6-腺苷甲基化(M6A)，以确定可能提供 对导致β细胞死亡的过程的重要线索。在本提案中，我们将：1)。描述一下 从1型糖尿病模型中获得的胰岛细胞中的动态RNA甲基化变化；2)确定 人类胰岛和β细胞中N6-甲基腺苷(M6A)变化的功能相关性；以及3)探索 T1D小鼠模型胰岛细胞RNA中额外甲基标记的变化最后，我们会 T1D模型胰岛RNA甲基化变化与蛋白质组学和基因对照研究 从T1D患者的胰岛中获得表达特征。

PROJECT SUMMARY The number of people with diabetes worldwide is predicted to increase from 415 million in 2015 to 642 million in 2040. The health care costs of treating the disease account for 12% of the global health expenditure and continue to be a huge economic burden (IDF World Atlas 2015). Type 1 diabetes mellitus (T1DM) constitutes 10-15% of the disease burden and is an autoimmune disease, thought to be triggered by genetic or environmental factors in early childhood, which results in antibody-mediated destruction of insulin producing pancreatic β-cells causing life-threatening hyperglycemia. In contrast, type 2 diabetes mellitus (T2DM) develops later in life and results from insulin resistance in target tissues and inflammation coupled with an inadequate compensation by the β-cells. The precise mechanisms that trigger the cascade of events ultimately leading to beta cell death are still not fully understood. Several observations have prompted renewed interest in the beta cell itself as a target tissue that could initiate the disease process. In this context the recent identification of RNA methylation as a potential regulatory mechanism that contributes to the ability of a cell to adapt to rapid changes in the environment provides a provocative approach to investigate changes in beta cells that precede the development of the type 1 diabetes phenotype. We seek to interrogate alterations in the dynamic methylation of RNA, specifically, N6-adenosine methylation (m6A) in beta cells to identify the signatures that might provide important clues to processes that contribute to beta cell death. In this proposal we will: 1). Characterize the dynamic RNA methylation changes in islet cells obtained from models of type 1 diabetes; 2) determine the functional relevance of alterations in N6-methyladenosine (m6A) in human islets and β-cells; and 3) explore changes in additional methyl marks in RNA in islet cells from models of T1D mouse model. Finally, we will contrast the RNA methylation changes observed in the islets in the T1D models with proteomics and gene expression signatures in islets obtained from patients with T1D.