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

项目摘要 全球糖尿病患者人数预计将从2015年的4.15亿增加到6.42亿 在2040年治疗这种疾病的卫生保健费用占全球卫生支出的12%， 继续是一个巨大的经济负担（IDF世界地图集2015）。1型糖尿病（T1 DM）是一种 10-15%的疾病负担，是一种自身免疫性疾病，被认为是由遗传或 环境因素在儿童早期，这导致抗体介导的破坏胰岛素产生 胰腺β细胞导致危及生命的高血糖症。与此相反，2型糖尿病（T2 DM） 在生命的后期发展，由靶组织中的胰岛素抵抗和炎症以及 β细胞的补偿不足。 触发最终导致β细胞死亡的级联事件的精确机制仍然存在 没有完全理解。几个观察结果促使人们重新对β细胞本身作为靶点产生兴趣 可能引发疾病的组织在这种情况下，最近确定的RNA甲基化作为一个 潜在的调节机制，有助于细胞适应快速变化的能力， 环境提供了一种挑衅性的方法来研究β细胞的变化， 1型糖尿病的发病机制我们试图询问动态甲基化的改变， RNA，特别是β细胞中的N6-腺苷甲基化（m6 A），以确定可能提供 这是导致β细胞死亡的重要线索在本提案中，我们将：1）。表征 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 人胰岛和β细胞中N6-甲基腺苷（m6 A）改变的功能相关性;以及3）探索 来自T1 D小鼠模型的胰岛细胞中RNA中额外甲基标记的变化。最后我们将 将T1 D模型中胰岛中观察到的RNA甲基化变化与蛋白质组学和基因组学进行对比， 从患有T1 D的患者获得的胰岛中的表达特征。

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.