Screens for novel compounds to correct diabetic postprandial dyslipidemia

筛选纠正糖尿病餐后血脂异常的新型化合物

• 批准号: 7651625
• 负责人: Kevin Jon Williams
• 金额: $ 37.5万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7651625
• 关键词: Action Potentials; Anabolism; Apolipoproteins B; Area; Atherosclerosis; Automation; Binding; Biological; Biological Assay; Biological Factors; Carbohydrates; Cardiovascular Diseases; Catabolism; Cause of Death; Cell surface; Cells; Cholesterol; Chylomicrons; Cleaved cell; Clinical Trials; Collaborations; Collection; Confusion; Cytochalasin D; Data; Deacetylase; Defect; Development; Diabetes Mellitus; Dyslipidemias; Employee Strikes; Enzymes; Ethnopharmacology; Evaluation; FDA approved; Fluorescence; Funding; Genistein; Goals; Head; Heparan Sulfate Proteoglycan; Heparin; Heparitin Sulfate; Hepatic; Hepatocyte; Human; Hypertriglyceridemia; Inorganic Sulfates; Insulin-Dependent Diabetes Mellitus; Intestines; Knock-out; Knowledge; Label; Laboratories; Length; Letters; Libraries; Life; Ligand Binding Domain; Lipoproteins; Liver; Liver Circulation; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Mediating; Mediator of activation protein; Metabolic; Methods; Miniaturization; Modeling; Molecular; Molecular Mechanisms of Action; Molecular Target; Morphologic artifacts; Mus; NIH Program Announcements; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Participant; Pathway interactions; Patients; Peripheral; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Plasma; Polymers; Preclinical Drug Evaluation; Preparation; Primary carcinoma of the liver cells; Proteins; Public Health; Publications; Publishing; Radioactive; Reading; Reagent; Reproducibility; Resources; Role; Route; Screening procedure; Seminal; Side; Specific qualifier value; Structure; Surface; Syndrome; Testing; Therapeutic; Tissues; Titrations; Triglycerides; Unspecified or Sulfate Ion Sulfates; Validation; Work; apolipoprotein B receptor; apolipoprotein B-48; assay development; base; cardiovascular risk factor; diabetic; diabetic patient; expectation; heparanase; heparitinase; high throughput screening; in vivo; insight; knock-down; lipoprotein cholesterol; neglect; non-diabetic; novel; novel strategies; particle; polypeptide; public health relevance; receptor; response; restoration; small molecule; sugar; sulfation; sulfotransferase; type I and type II diabetes; uptake

DESCRIPTION (provided by applicant): Atherosclerotic cardiovascular disease remains the major cause of death in patients with type 1 and type 2 diabetes mellitus (T1DM, T2DM). Atherosclerosis arises from the retention of cholesterol-rich, apolipoprotein-B (apoB)-lipoproteins within the vessel wall. Importantly, diabetic patients suffer from a unique and typically neglected aspect of cardiovascular risk, namely, the striking persistence of intestinally derived apoB-lipoproteins, called `remnants,' in their plasma after each meal. The cause is a defect in hepatic clearance of these harmful particles. A major impediment in this area has been our ignorance regarding pathways for remnant uptake into the liver. A quarter century ago, hepatic uptake of remnants was shown to be independent of LDL receptors. This realization launched a long, difficult search for the responsible molecules. In 1991-1992, seminal work from our laboratory implicated heparan sulfate proteoglycans (HSPGs) in remnant lipoprotein uptake. Each HSPG molecule consists of a protein strand onto which the cell assembles sugar polymers, called heparan sulfate, that we showed could capture lipoproteins. In a major, recent breakthrough, we found that T2DM induces HSPG degradative enzymes in liver. Our central hypothesis is that the identification of compounds that accelerate uptake of atherogenic remnant lipoproteins into liver cells will substantially advance our physiologic understanding while also opening exciting, new avenues for potentially life-saving therapeutics in diabetes. In Aim 1, we propose to develop screens for inducers of hepatocyte uptake of model remnant lipoproteins. Compound screens using whole-cell read-outs are the ideal approach to manage the biologic complexity of HSPG assembly. Aim 1a will automate and optimize our new fluorescent assay for HSPG-mediated uptake of model remnant lipoproteins, in preparation for moderate-throughput screening. Aim 1b will perform titration-based screening of three libraries of diverse, active compounds. In Aim 2, we propose to evaluate hit compounds. Aim 2a will rule out artifacts, establish selectivity for HSPG-mediated uptake, and prioritize compounds. Aim 2b will determine the molecular effects of prioritized compounds. Based on our new data, our favored mechanism is inhibition of HSPG degradative enzymes. Aim 2c will define metabolic responses to enhanced uptake of remnants, to choose potential therapeutic leads. Overall, these proposed Aims will substantially advance our molecular knowledge of remnant lipoprotein clearance, as well as our ability to correct diabetic postprandial dyslipidemia. Our ultimate goal will be to avert the tremendous excess burden of cardiovascular disease in diabetes, to which postprandial dyslipidemia makes a substantial, and potentially avoidable, contribution. PUBLIC HEALTH RELEVANCE: Project relevance to public health Patients with diabetes mellitus suffer from fatal and disabling atherosclerotic cardiovascular disease that results in part from the striking persistence of harmful intestinally derived lipoproteins, called `remnants,' in their plasma after each meal. Based on our seminal work implicating a crucial role for heparan sulfate proteoglycans (HSPGs) in the rapid, healthy disposal of remnant lipoproteins by the liver, we now seek novel compounds to enhance HSPG display by hepatocytes and thereby accelerate the uptake of these harmful lipoproteins. The ultimate goal will be to avert the tremendous excess burden of cardiovascular disease in diabetes.

描述（由申请方提供）：动脉粥样硬化性心血管疾病仍然是1型和2型糖尿病（T1 DM、T2 DM）患者的主要死亡原因。动脉粥样硬化是由富含胆固醇的载脂蛋白B（apoB）-脂蛋白在血管壁内的滞留引起的。重要的是，糖尿病患者患有心血管风险的一个独特且通常被忽视的方面，即，在每餐后血浆中显著持续存在来自母体的apoB-脂蛋白，称为“残余物”。原因是肝脏对这些有害颗粒的清除存在缺陷。这一领域的一个主要障碍是我们对残留物进入肝脏的途径的无知。四分之一世纪前，肝脏对残余物的摄取被证明不依赖于LDL受体。这一认识启动了对责任分子的长期而艰难的搜索。在1991-1992年，我们实验室的开创性工作涉及硫酸乙酰肝素蛋白聚糖（HSPGs）在残余脂蛋白摄取。每个HSPG分子由一条蛋白链组成，细胞在其上组装糖聚合物，称为硫酸乙酰肝素，我们发现它可以捕获脂蛋白。在最近的一项重大突破中，我们发现T2 DM诱导肝脏中的HSPG降解酶。我们的中心假设是，识别出加速肝细胞吸收致动脉粥样硬化残余脂蛋白的化合物将大大提高我们的生理认识，同时也为糖尿病潜在的救生治疗开辟令人兴奋的新途径。在目的1中，我们建议开发模型残余脂蛋白的肝细胞摄取的诱导剂的筛选。使用全细胞读数的化合物筛选是管理HSPG组装的生物复杂性的理想方法。目标1a将自动化和优化我们的新的荧光检测HSP介导的模型残余脂蛋白的摄取，在中等通量筛选的准备。目标1b将对三个不同的活性化合物库进行基于滴定的筛选。在目标2中，我们建议评估命中化合物。目标2a将排除伪影，建立对HSPG介导的摄取的选择性，并优先考虑化合物。目标2b将确定优先化合物的分子效应。根据我们的新数据，我们的有利机制是抑制HSPG降解酶。目标2c将定义代谢反应，以增强残留物的摄取，选择潜在的治疗线索。总之，这些提出的目标将大大提高我们对残余脂蛋白清除的分子知识，以及我们纠正糖尿病餐后血脂异常的能力。我们的最终目标将是避免糖尿病患者心血管疾病的巨大过度负担，餐后血脂异常对糖尿病患者心血管疾病的负担有很大的影响，而且可能是可以避免的。公共卫生关系：糖尿病患者患有致命和致残的动脉粥样硬化性心血管疾病，部分原因是每餐后血浆中有害的来源于母体的脂蛋白（称为“残余物”）的惊人持久性。基于我们的开创性工作，涉及硫酸乙酰肝素蛋白聚糖（HSPG）在肝脏对残余脂蛋白的快速，健康处置中的关键作用，我们现在寻求新的化合物来增强肝细胞对HSPG的展示，从而加速这些有害脂蛋白的摄取。最终目标将是避免糖尿病心血管疾病的巨大过度负担。