MDM2-p53信号通路调控胰岛β细胞胰岛素分泌功能的机制研究

项目名称： MDM2-p53信号通路调控胰岛β细胞胰岛素分泌功能的机制研究

项目编号： No.81500624

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 李晓牧

作者单位： 复旦大学

项目金额： 18万元

中文摘要： 胰岛β细胞功能异常与2型糖尿病密切相关,但其调控机制尚不明确。MDM2-p53信号通路参与细胞应激和葡萄糖代谢的调控机制，迄今对其与β细胞功能的关系知之甚少。我们的预实验结果发现肥胖小鼠胰岛β细胞MDM2，p53水平显著上调；MDM2抑制剂导致β细胞p53上调，胰岛素分泌缺陷和线粒体功能异常。因此我们提出假说：MDM2-p53可能通过抑制线粒体代谢影响β细胞胰岛素分泌功能。为验证这一假说，我们将利用已建立的MDM2胰岛β细胞选择性敲除小鼠、离体胰岛、及MIN6 β细胞为研究模型，通过胰岛素分泌功能测定、线粒体功能测定、慢病毒感染系统、生物信息学等手段，从分子、细胞、组织以及动物水平等层次探讨MDM2-p53对β细胞分泌功能的调控作用， 明确MDM2-p53通过对PC转录调控影响线粒体功能的机制。本研究将从MDM2-p53这个新视点揭示调控β细胞功能的基本机制，为防治2型糖尿病提供新靶点。

中文关键词： 2型糖尿病；胰岛素分泌；胰岛；β；细胞；MDM2-p53；通路

英文摘要： Type 2 diabetes mellitus (T2DM) and its complications are becoming a major global health burden. Defective glucose-stimulated insulin secretion (GSIS) in pancreatic β-cells is a key feature of T2DM, and contributes to the development of hyperglycemia. Although chronic metabolic stress (eg. obesity or nutrient overload) has been shown to impair β-cell function, the underlying mechanism remains unclear. A thorough understanding of the pathogenesis of β-cell failure will help to develop more therapeutic strategies against diabetes by relieving metabolic stress in β-cells. The tumor suppressor p53 is stabilized and becomes transcriptionally active in response to genotoxic stress, thereby controlling cell cycle. Apart from these established roles, p53 has recently been shown to contribute to the development of metabolic disorders such as insulin resistance and premature aging. Recently, we showed that p53 is stabilized and activated in β-cells of obese mice with β-cell dysfunction, which is accompanied with an elevation of mouse double minute 2 (MDM2), a principle negative regulator of p53. Treatment with the MDM2 inhibitor Nutlin-3a, which stabilizes and activates p53 by disrupting the interaction between MDM2 and p53, causes defective GSIS in β-cell. Such a defect is perhaps due to aberrant tricarboxylic acid (TCA) cycle and diminished ATP production in mitochondria. Based on these results, we proposed that a tight control of the MDM2-p53 axis is important for maintaining β-cell function. Chronic metabolic stress activates p53, leading to aberrant mitochondrial functions, thereby resulting in β-cell dysfunction and T2DM. In this project, we will test whether genetic deletion of MDM2 in β-cells results in an uncontrolled activation of p53, leading to defective GSIS and glucose intolerance, prove the concept that chronic activation of p53 by deleting its negative regulator MDM2 leads to the onset of T2DM, and inactivation of p53 in β-cells can alleviate MDM2 deletion induced deleterious effects on β-cell function. In addition, we will investigate how the interplay between p53 and MDM2 fine-tune GSIS by modulating TCA cycle and ATP production via a novel p53 target pyruvate carboxylase (PC) gene. The results will provide new insight into the fundamental mechanisms of β-cell function and regulation of insulin secretion, and to shed new light on the pathogenesis of glucose dysregulation in diabetes. The unique mouse model established in this study will serve as a useful tool to identify and validate novel drug targets for the development of therapeutics to combat diabetes, one of the major chronic disease in China.

英文关键词： Type 2 Diabetes;Insulin secretion;β cell;MDM2-p53 pathway