Glucokinase Regulation of Hepatic Metabolism

项目摘要 我的职业目标是领导一个生物医学研究小组，研究潜在的分子原因 糖尿病和肥胖症等胰岛素抵抗状态下肝功能障碍的机制。要获得 具体的培训我将需要实现这一点，我提出了一个项目，使用最先进的分子和 用生化方法研究激素和营养物质(如胰岛素和葡萄糖)是如何协调的 餐后状态下的肝脏代谢以及这些过程如何在代谢过程中被破坏 疾病。我的职业发展计划将扩展我在激素信号和 代谢生理学，并提供全基因组转录和生物信息学分析方面的基本培训， 和蛋白质生物化学。因为我的目标是确定新的转录和信号机制， 调节肝脏代谢，我作为一名独立研究人员的成功依赖于这些技能。 目前的研究方案是研究肝脏葡萄糖激酶如何调节肝脏基因转录和 代谢以及这些过程如何导致糖尿病期间的代谢功能障碍。我已经开发出一种 新的小鼠模型，分离肝脏对胰岛素和葡萄糖的反应，这是定义 每种信号在调节肝脏代谢中的具体作用。葡糖激酶在肝脏中的特异性表达 缺乏肝脏Akt亚型的小鼠足以恢复正常的糖耐量并进行“经典”调节。 定义了胰岛素反应基因。这些数据表明，葡萄糖信号的增加绕过了这一要求 胰岛素在肝脏中的直接作用，以调节转录和代谢。在这个被指导的职业生涯中 获得发展奖后，我将获得全基因组转录和蛋白质调控方面所需的专业知识 这些技术在我导师的实验室里已经很成熟了。我将检验有缺陷的假设 葡萄糖激酶对葡萄糖的利用导致转录调节异常，导致糖耐量异常 和胰岛素抵抗。使用在此K01应用程序中学到的最先进的技术，我将定义 在饮食过程中导致肝脏代谢改变的全肝基因组转录机制。 诱导肥胖并检测转录因子Foxo1在介导这些过程中的需求。 此外，我将正式描述肝脏葡萄糖利用减少和增加之间的关系 Foxo1活性在胰岛素抵抗发生发展中的作用。最后，我将利用我新学到的专业知识 转录分析，以确定调控机制和转录因子，除了Foxo1， 负责协调肝脏对营养摄入的转录反应。这项指导支持 再加上我现有的专业知识，将为成功的独立学术生涯提供框架。

Project Summary My career goal is to lead a biomedical research group that investigates the causal underlying molecular mechanisms of hepatic dysfunction during insulin-resistant states such as diabetes and obesity. To acquire the specific training I will need to achieve this, I propose a project that uses state-of-the-art molecular and biochemical approaches to investigate how hormones and nutrients such as insulin and glucose coordinate hepatic metabolism in the postprandial state and how these processes become disrupted during metabolic disease. My career development plan will expand upon my primary expertise in hormone signaling and metabolic physiology and provide essential training in genome-wide transcriptional and bioinformatic analysis, and protein biochemistry. Since it is my goal to identify novel transcriptional and signaling mechanisms that regulate hepatic metabolism, my success as an independent researcher depends on these skills. The current research proposal is to study how hepatic glucokinase regulates liver gene transcription and metabolism and how these processes contribute to metabolic dysfunction during diabetes. I have developed a novel mouse model that dissociates the hepatic response to insulin and glucose, which is critical to define the specific contribution of each signal in regulating liver metabolism. Liver-specific expression of glucokinase in mice lacking hepatic Akt isoforms was sufficient to normalize glucose tolerance and to regulate “classically” defined insulin responsive genes. These data suggest increased glucose signaling bypasses the requirement of direct insulin action in liver to regulate transcription and metabolism. During this mentored career development award, I will gain the required expertise in genome-wide transcriptional and protein regulation techniques that are well established in my mentors' laboratories. I will test the hypothesis that defective glucose utilization by glucokinase leads to aberrant transcriptional regulation resulting in glucose intolerance and insulin resistance. Using state-of-the-art techniques learned during this K01 application, I will define the hepatic genome-wide transcriptional mechanisms that contribute to altered hepatic metabolism during diet- induced obesity and test the requirement of the transcription factor Foxo1 in mediating these processes. Moreover, I will formally delineate the relationship between reduced hepatic glucose utilization and increased Foxo1 activity in the development of insulin resistance. Finally, I will use my newly acquired expertise in transcriptional analysis to define the regulatory mechanisms and transcription factors, in addition to Foxo1, responsible for coordinating the hepatic transcriptional response to nutrient intake. This mentoring support coupled with my existing expertise will provide the framework for a successful independent academic career.