Probing Diabetes: Development of a HTS-compatible TORC2 Redistribution Assay.

探索糖尿病：开发兼容 HTS 的 TORC2 再分布测定。

• 批准号: 7727585
• 负责人: Michael Dale Conkright
• 金额: $ 34.04万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7727585
• 关键词: Address; Adipose tissue; Affect; Agonist; Attenuated; Biological Assay; Biological Process; Blood Glucose; CREB1 gene; Cell Nucleus; Cells; Chemicals; Chronic; Chronic Disease; Complex; Cyclic AMP; Cytoplasm; Development; Diabetes Mellitus; Diabetic mouse; Disabled Persons; Discrimination; Disease; Etiology; Fasting; Florida; Funding; Funding Opportunities; Gene Expression; Genes; Glucagon; Gluconeogenesis; Goals; Health; Hepatic; Hepatocyte; Hormones; Hyperglycemia; Individual; Insulin; Lead; Libraries; Liver; Mediating; Mission; Molecular; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Export; Nuclear Import; Nuclear Translocation; Pathway interactions; Phosphoenolpyruvate Carboxylase; Phosphorylation; Physiological; Preclinical Drug Evaluation; Proteins; Reagent; Reproducibility; Research Institute; Resources; Screening procedure; Signal Pathway; Signal Transduction; Skeletal Muscle; Therapeutic; United States; United States National Institutes of Health; assay development; base; blood glucose regulation; counterscreen; design; feeding; glucose output; glucose production; glucose uptake; hepatic gluconeogenesis; high throughput screening; innovation; insulin signaling; miniaturize; mouse model; novel therapeutics; pandemic disease; programs; public health relevance; response; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): Type II diabetes is simply characterized by hyperglycemia but is a disease state with a complex etiology. In addition to decreased glucose uptake by adipose and skeletal muscle, chronic hepatic glucose production contributes to the manifestation of this disease and is the central health related problem addressed by the proposed studies. Glucagon initiates the gluconeogenic program by activating the cAMP-signaling cascade that is, in part, mediated by the nuclear translocation of the CREB coactivator, TORC2. Indeed, the redistribution of TORC2 is a key regulatory step in glucose homeostasis and small molecules that are capable of blocking nuclear import of TORC2, or that facilitate its nuclear export, are likely to regulate hepatic glucose production. The goal of this project is to develop and validate a TORC2 subcellular localization assay that is amenable to high-throughput screening of large chemical libraries, with a long-term goal of identifying small molecules that target TORC2 redistribution and regulate hepatic gluconeogenesis. To this end, a portfolio of confirmatory screens will also be developed in parallel, which will discriminate small molecules producing false- positive results during the primary screening campaign. Finally, a cell-based gluconeogenesis assay will also be developed to validate the physiological relevance of small molecules hits coming from our primary and secondary screens. Collectively, the assays developed in the proposed studies will provide the toolsets necessary for a high-throughput screening campaign, to fulfill the long-term goal of identifying small molecule chemical probes that will aid in our understanding of aberrant gluconeogenesis, and that may lead to novel therapeutic molecules for the treatment of type II diabetes. PUBLIC HEALTH RELEVANCE: Characterized by high blood glucose, type II diabetes has reached pandemic proportions with approximately 20 million individuals affected in the United States alone. Contributing to the manifestation of this disease, chronic glucose production by the liver is one component of diabetes that is mediated by the cAMP-signaling pathway. By search for small molecules capable of blocking this signaling pathway we will uncover chemical probes that will facilitate our understanding this disease and will possibly function as novel therapeutic molecules for the treatment of individuals suffering from diabetes.

描述(申请人提供)：II型糖尿病的简单特征是高血糖，但它是一种具有复杂病因的疾病状态。除了脂肪和骨骼肌对葡萄糖的摄取减少外，慢性肝糖的产生也是这种疾病的表现，也是拟议研究解决的与健康有关的中心问题。胰高血糖素通过激活cAMP信号级联来启动糖异生程序，cAMP信号级联在一定程度上是由CREB辅活化子TORC2的核转位介导的。事实上，TORC2的重新分布是葡萄糖稳态的关键调控步骤，能够阻止TORC2核输入或促进其核输出的小分子可能调节肝脏葡萄糖的产生。该项目的目标是开发和验证一种TORC2亚细胞定位分析，该方法适用于对大型化学库的高通量筛选，长期目标是识别靶向TORC2再分布和调节肝脏糖异生的小分子。为此，还将同时开发一系列确认性筛查，这些筛查将区分在初步筛查活动中产生假阳性结果的小分子。最后，还将开发一种基于细胞的糖异生试验，以验证来自我们的初级和次级筛选的小分子点击的生理学相关性。总之，拟议研究中开发的分析方法将为高通量筛查活动提供必要的工具集，以实现识别小分子化学探针的长期目标，这些小分子化学探针将有助于我们了解异常的糖异生作用，并可能导致治疗II型糖尿病的新型治疗分子。公共卫生相关性：以高血糖为特征的II型糖尿病已经达到大流行的程度，仅在美国就有大约2000万人受到影响。肝脏产生的慢性葡萄糖是糖尿病的一个组成部分，它是由cAMP信号通路介导的，是导致这种疾病的原因之一。通过寻找能够阻断这一信号通路的小分子，我们将发现有助于我们理解这种疾病的化学探针，并可能作为新的治疗分子用于糖尿病患者的治疗。