Membrane-Based Mechanisms of Hepatic Insulin Resistance

肝胰岛素抵抗的膜机制

• 批准号: 9350309
• 负责人: Baran Ersoy
• 金额: $ 15.19万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9350309
• 关键词: Ablation; Activation Analysis; Advisory Committees; Affect; Applications Grants; Attenuated; Automobile Driving; Basic Science; Binding; Binding Proteins; Biological; Bone Marrow Transplantation; Calcium Channel; Cell Culture System; Cell Culture Techniques; Cells; Cellular biology; Chemicals; Committee Members; Data; Defect; Development; Diabetes Mellitus; Diet; Disease; Education; Endocrinologist; Endoplasmic Reticulum; Environment; Enzyme-Linked Immunosorbent Assay; Exhibits; Extrahepatic; Fasting; Genetic; Glucose Metabolism Disorders; Goals; Harvest; Hepatic; Hepatocyte; Hepatology; High Fat Diet; Hospitals; IRS2 gene; Immune response; Immunoblot Analysis; Impairment; In Vitro; Inflammation; Inflammatory; Insulin; Insulin Receptor; Insulin Resistance; Interleukin-6; Intervention; Laboratories; Lead; Lecithin; Link; Lipid Binding; Lipids; Lipopolysaccharides; Liver; MAPK8 gene; Macrophage Activation; Mass Spectrum Analysis; Measures; Mediating; Medication Management; Medicine; Membrane; Mentored Research Scientist Development Award; Mentors; Mentorship; Metabolic; Mission; Molecular; Molecular Biology; Molecular Immunology; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Outcome Study; Pathway interactions; Peritoneal Macrophages; Pharmacology; Phospholipids; Phosphotransferases; Physician Executives; Physiological; Play; Positioning Attribute; Prevention; Proteins; Proteomics; Public Health; RNA Splicing; Reagent; Regulation; Research; Research Personnel; Research Proposals; Resistance; Role; Scientist; Signal Transduction; Small Interfering RNA; Starvation; TNF gene; Techniques; Testing; Time; Tissues; Training; Transgenic Mice; Tunicamycin; Woman; base; biological adaptation to stress; blood glucose regulation; career; common treatment; cytokine; endoplasmic reticulum stress; experience; experimental study; feeding; glucose metabolism; glucose production; improved; instructor; insulin sensitivity; insulin signaling; knock-down; lipid metabolism; loss of function; medical schools; mouse model; new therapeutic target; non-alcoholic fatty liver; novel; novel therapeutic intervention; phosphatidylcholine transfer protein; professor; protein complex; public health relevance; response; skills; small molecule; small molecule inhibitor; success

DESCRIPTION (provided by applicant): This is an application for a K01 Award for Dr. Baran A. Ersoy, an Instructor in Medicine at the Brigham and Women's Hospital (BWH) and Harvard Medical School (HMS). Dr. Ersoy is a young investigator in basic research on disorders of glucose and lipid metabolism. His application outlines a short-term plan to further his education by gaining experience in advanced molecular biology and immunology techniques, including mass spectrometry of proteomics and bone marrow transplantation. This plan will build upon his previous experience and provide him the time and training to establish new research skills, strategies and projects that will contribute to his success upon becoming an independent scientist. It is his long-term goal to become an establish independent investigator in an academic setting and study the mechanisms driving insulin resistance and promoting type 2 diabetes and non-alcoholic fatty liver disease (NAFLD). To achieve these goals, Dr. Ersoy will be mentored by Dr. David E. Cohen, Professor of Medicine at HMS and Director of Hepatology at BWH. Dr. Ersoy has also assembled an advisory committee comprised of Dr. Steven P. Gygi, Professor of Cell Biology at HMS, who focuses on mass spectrometry of proteomics; Dr. Umut Ozcan, an endocrinologist with expertise in endoplasmic reticulum stress, inflammation and insulin signaling; and Dr. Morris F. White, an expert in insulin signaling who played a major role in the discovery insulin receptor substrate proteins. Dr. Ersoy's mentor and advisory committee members have established mentorship and training record, whose previous trainees hold independent investigator positions throughout the world. Dr. Ersoy will conduct his training at BWH and HMS, which will provide an intellectually rich research environment and has a long- standing commitment to training young scientists for independent academic research careers. Obesity-induced resistance to insulin action is the primary pathophysiological defect that predisposes to NAFLD and type 2 diabetes. Because current management options remain limited, identification of new regulatory mechanisms that govern the metabolic response to insulin should serve to identify novel opportunities for pharmacologic intervention. The objective of this research proposal is to elucidate a fundamental new mechanism for the regulation of glucose homeostasis. The rationale is that the identification of a novel mechanism that regulates hepatic insulin sensitivity could lead to the identification of new therapeutic targets for the treatment of NAFLD and type 2 diabetes. Guided by extensive preliminary data, the central hypothesis of this research plan is that phosphatidylcholine transfer protein (PC-TP) regulates glucose homeostasis by controlling the activity of insulin receptor substrate 2 (IRS2) both directly and also indirectly by modulating endoplasmic reticulum (ER) stress and inflammation. This will be tested in three specific aims: 1) Define the mechanisms by which PC-TP suppresses IRS2 activity; 2) Determine the role of ER stress in PC- TP-mediated regulation of insulin signaling; 3) Elucidate the role of immune response in PC-TP-mediated inhibition of insulin sensitivity. In Aim 1, a small molecule inhibitor of PC-TP (compound A1) and IRS2 deficient mice will be used to examine the role of IRS2 in the regulation of insulin signaling by PC-TP. The biological pathways in the control of IRS2 inhibition by PC-TP will be unveiled using mass spectroscopy of proteomics in Pctp+/+ and Pctp-/- mice as well as compound A1-treated hepatocytes harvested from these mice. In Aim 2, Pctp+/+ and Pctp-/- mice and hepatocytes harvested from these mice will be used to determine whether PC-TP modulates key ER stress pathways that regulate insulin signaling and IRS2 activity by immunoblot analyses. ER stress signaling will be activated by refeeding following 24 h starvation in mice, and by the chemical reagent tunicamycin in cell culture systems. Aim 3 will utilize peritoneal macrophages harvested from Pctp+/+ and Pctp-/- mice and bone marrow transplantation of Pctp-/- donor cells into Pctp+/+ recipient mice. Inflammation will be induced by high fat diet feeding in mice, and by lipopolysaccharide treatment in cell culture. Inflammatory signals will be measured by ELISA, immunoblot and qPCR analyses. It is anticipated that PC-TP will regulate insulin signaling by regulating IRS2 activity both directly and by modulating ER stress and inflammation. This is important because loss of IRS2 expression and increased ER stress and inflammation are associated with development of insulin resistance. These studies are expected to identify new therapeutic targets for the management of insulin resistance and related disorders. The data generated from this research proposal will form the basis for an R01 grant application at the end of the K01 award.

描述(由申请人提供)：这是一份K01奖的申请书，授予Baran A.Ersoy博士，她是布里格姆妇女医院(BWH)和哈佛医学院(HMS)的医学讲师。Ersoy博士是糖脂代谢紊乱基础研究领域的一名年轻研究员。他的申请勾勒出了一个短期计划，通过获得先进的分子生物学和免疫学技术的经验，包括蛋白质组学和骨髓移植的质谱学，来继续他的教育。这项计划将以他以前的经验为基础，并为他提供时间和培训，以建立新的研究技能、战略和项目，这些将有助于他成为一名独立科学家后取得成功。他的长期目标是成为一名学术背景下的独立研究者，研究推动胰岛素抵抗和促进2型糖尿病和非酒精性脂肪性肝病(NAFLD)的机制。为了实现这些目标，Ersoy博士将得到HMS医学教授兼BWH肝病主任David E.Cohen博士的指导。Ersoy博士还组建了一个顾问委员会，成员包括HMS细胞生物学教授Steven P.Gygi博士，他专注于蛋白质组学的质谱学；内分泌学家Umut Ozcan博士，他在内质网应激、炎症和胰岛素信号方面具有专长；以及胰岛素信号专家莫里斯·F·怀特博士，他在胰岛素受体底物蛋白的发现中发挥了重要作用。Ersoy博士的导师和咨询委员会成员已经建立了导师关系和培训记录，他们之前的受训人员在世界各地担任独立研究员的职位。Ersoy博士将在BWH和HMS进行他的培训，这将提供一个智力丰富的研究环境，并长期致力于为独立的学术研究事业培养年轻科学家。肥胖引起的胰岛素抵抗是NAFLD和2型糖尿病的主要病理生理缺陷。由于目前的管理选择仍然有限，识别管理胰岛素代谢反应的新调节机制应该有助于识别药物干预的新机会。这项研究计划的目的是阐明一种基本的新机制来调节葡萄糖的动态平衡。其基本原理是，对调节肝脏胰岛素敏感性的新机制的识别可能导致识别治疗NAFLD和2型糖尿病的新治疗靶点。在大量前期数据的指导下，本研究计划的中心假设是磷脂酰胆碱转移蛋白(PC-TP)通过直接和间接地通过调节内质网(ER)应激和炎症来控制胰岛素受体底物2(IRS2)的活性来调节血糖稳态。这将在三个特定的目标中得到验证：1)明确PC-TP抑制IRS2活性的机制；2)确定内质网应激在PC-TP介导的胰岛素信号调节中的作用；3)阐明免疫反应在PC-TP介导的胰岛素敏感性抑制中的作用。在目标1中，将使用PC-TP的小分子抑制剂(化合物A1)和IRS2缺陷小鼠来研究IRS2在PC-TP调节胰岛素信号转导中的作用。在Pctp+/+和Pctp-/-小鼠以及从这些小鼠收集的化合物A1处理的肝细胞中，将使用蛋白质组学的质谱学揭示PC-TP控制IRS2抑制的生物学途径。在目标2中，Pctp+/+和Pctp-/-小鼠以及从这些小鼠收集的肝细胞将被用来通过免疫印迹分析来确定PC-TP是否调节调节胰岛素信号和IRS2活性的关键ER应激通路。在小鼠饥饿24小时后的再喂养和细胞培养体系中的化学试剂衣霉素可以激活内质网应激信号。目的3将利用从Pctp+/+和Pctp-/-小鼠体内获取的巨噬细胞和将Pctp-/-供体细胞移植到Pctp+/+受体小鼠体内。高脂饲料喂养小鼠可引起炎症反应，细胞培养时可用脂多糖处理。炎症信号将通过酶联免疫吸附试验、免疫印迹和定量聚合酶链式反应检测。预计PC-TP将通过直接调节IRS2活性和通过调节内质网应激和炎症来调节胰岛素信号。这一点很重要，因为IRS2表达的丧失以及内质网应激和炎症的增加与胰岛素抵抗的发展有关。这些研究有望为胰岛素抵抗和相关疾病的管理确定新的治疗目标。这项研究提案产生的数据将成为K01奖项结束时R01拨款申请的基础。