Molecular Regulation of HDL Metabolism and Reverse Cholesterol Transport

HDL 代谢和反向胆固醇转运的分子调控

• 批准号: 7596525
• 负责人: Daniel James Rader
• 金额: $ 34.2万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596525
• 关键词: Address; Adopted; Affect; Animals; Antiatherogenic; Apolipoproteins A; Atherosclerosis; Biological; Biological Assay; Blood Vessels; Budgets; C-terminal; Cardiovascular Diseases; Cardiovascular system; Carrier Proteins; Categories; Cell Line; Cells; Chemicals; Cholesterol; Cholesterol Esters; Clinical; Collaborations; Complement; Coronary Arteriosclerosis; Critiques; Data; Development; Diet; Direct Costs; Equipment; Esterification; Excretory function; Face; Gene Expression; Goals; Health Insurance; Hepatic; High Density Lipoprotein Cholesterol; High Density Lipoproteins; Human; Human Resources; Hybrids; Hydrophobicity; In Vitro; Inpatients; Institutes; Instruction; Investigation; Investigational Drugs; Knock-out; Knockout Mice; Last Name; Life; Link; Lipid Binding; Lipoproteins; Liver; Low Density Lipoprotein Receptor; Measures; Mediating; Medicine; Metabolism; Methods; Modeling; Molecular; Molecular and Cellular Biology; Mus; Mutation; N-terminal; Names; Outpatients; Pathology; Pathway interactions; Patient Care; Pennsylvania; Peripheral; Pharmacology; Phenotype; Philadelphia; Phosphatidylcholine-Sterol O-Acyltransferase; Phospholipid Transfer Proteins; Physiology; Phytosterols; Plasma; Plastics; Play; Postdoctoral Fellow; Principal Investigator; Process; Property; Protein Inhibition; Proteins; Publications; Published Comment; Publishing; Radiolabeled; Reagent; Registries; Regulation; Research; Research Personnel; Reticuloendothelial System; Risk; Role; Schools; Specialist; Sterols; Structure; Tangier Disease; Therapeutic; Time; Tissues; Tracer; Travel; Universities; Variant; Wages; Wild Type Mouse; Work; atherogenesis; base; cardiovascular risk factor; cost; expression vector; high density lipoprotein-2; human embryonic stem cell; in vivo; in vivo Model; inhibitor/antagonist; insight; intravenous injection; lecithin cholesterol acyltransferase deficiency; macrophage; medical schools; mortality; mouse model; novel; novel therapeutic intervention; overexpression; professor; programs; protein expression; radiochemical; radiotracer; receptor expression; research study; response; reverse cholesterol transport; species difference; tool; torcetrapib; translational medicine; uptake; vector

PROJECT 3: MOLECULAR REGULATION OF HDL METABOLISM AND REVERSE CHOLESTEROL TRANSPORT High density lipoproteins (HDL) and their major protein apolipoprotein A-l (apoA-l) are thought to protect against atherosclerotic cardiovascular disease at least in part by promoting reverse cholesterol transport (RCT), whereby excess cholesterol is removed from the periphery (such as the vascular wall) and returned to the liver for excretion. The broad goal of this project is to generate greater understanding of the molecular physiology of HDL metabolism as it relates to reverse cholesterol transport (RCT) by using integrated in vivo models in mice and extending experiments where possible into humans. It has become clear that the plasma concentration of HDL cholesterol (HDL-C) is not an adequate surrogate for anti-atherogenic effects, and that measures of cholesterol flux through the RCT pathway and of HDL function are more critical with regard to effects on cardiovascular disease. Specific Aim 1 will involve the use of mouse in vivo experiments to study the structure-function properties of apoA-l, in studies that complement the physico- chemical studies in Project 2 and the cell-based studies in Project 1. ApoA-l variants will be inserted into AAV8-based gene expression vectors, which will be used to express these variant apoA-l molecules in apoA-l knockout mice and effects on HDL metabolism, RCT, and in selected cases, atherosclerosis will be determined. Specific Aim 2 will address, using mouse models, the roles of lecithin:cholesterol acyltransferase (LCAT), cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP) in modulating the rate of macrophage RCT and the relationship to atherogenesis. Specific Aim 3 will extend concepts and approaches developed in mice into the human setting, with the specific goal of measuring RCT using new methods in humans with Tangier disease (ABCA1 deficiency) and with LCAT deficiency. While high levels of HDL cholesterol are associated with reduced risk of coronary artery disease, it is not clear that simply raising levels of HDL cholesterol are sufficient to reduce cardiovascular risk. The mechanisms by which HDL is altered and the impact on the process of reverse cholesterol transport is likely to be a more important determinant of cardiovascular risk. This project will seek to understand the regulation of reverse cholesterol transport in living mice and humans. University of Pennsylvania School of Medicine Institute for Translational Medicine and Therapeutics 654 BRBII/III 421 Curie Blvd Philadelphia, PA 19104 PHS 398 (Rev. 04/06) Form Page 2 173 Principal Investigator/Program Director (Last, First, Middle): Phillips, Michael C. PROJECT 3 KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator(s). List all other key personnel in alphabetical order, last name first. Name Rader, Daniel J. Professor of Medicine, Pathology and Pharmacology Cuchel, Marina Research Assistant Prof, of Medicine OTHER SIGNIFICANT CONTRIBUTORS Name Chaitanya Divgi Alan Remaley eRA Commons User Name Organization Role on Project Universityof DANRADER PennsylvaniaSchool of PI Medicine University of BILLHEIJ Universityof MCUCHEL Pennsylvania School of Co-Investigator Medicine Organization Role on Project University of Pennsylvania Collaborator NIH/NHLBI Collaborator Human Embryonic Stem Cells X No Q Yes If the proposed project Involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://Stemcells.nih.qov/reqistry/index.asp. Use continuation pages as needed. If a specificline cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line PHS 398 (Rev. 04/06) Form Page 2-continued 174 Principal Investigator / Program Director (Last. First. Middle): Phillips. Michael C. PROJECT 3 FROM THROUGH DETAILED BUDGET FOR INITIAL BUDGET PERIOD DIRECT COSTS ONLY 12/01/08 11/30/09 PERSONNEL (Applicant organization only) Months Devoted to Proiect ROLE ON Cal. Acad. Summer NAME PROJECT Mnths Mnths Mnths Principal Rader, Daniel Investigator 2.40 Billheimer, Jeffrey Co-Investigator 1.20 Cuchel, Marina Co-Investigator 0.6 Postdoctoral Tanigawa, Hiroyuki 12.0 Fellow Research Deborah Cromley 3.0 Specialist Research AishaWilson 6.0 Specialist Page 2 SUBTOTALS CONSULTANT COSTS EQUIPMENT (Itemize) SUPPLIES (Itemize) Plastic/Glassware Chemicals, radiochemicals and biological reagents Clinical supplies and investigational drugs Animal purchase TRAVEL PATIENT CARE COSTS INPATIENT OUTPATIENT ALTERATIONS AND RENOVATIONS (Itemize by category) OTHER EXPENSES (Itemize by category) Animal Per Diems Post doc health insurance Publications Subject Comoensation CONSORTIUM/CONTRACTUAL COSTS DOLLAR AMOUNT REQUESTED (omit cents) INST.BASE SALARY FRINGE SALARY REQUESTED BENEFITS TOTAL **** 37,320 11,159 48,479 97,600 9,760 2,918 12,678 86,570 0 0 0 55,426 55,426 5,376 60,802 62,588 15,647 4,678 20,325 43,894 21,947 6,562 28,509 26,252 7,850 34,102 ^w 166,352 38,543 204,895 7,013 10,000 4,000 10,000 31,013 10,000 8,100 2,500 12.500 33,100 DIRECT COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD (Item 7a. Face Page) $ 269 008 CONSORTIUM/CONTRACTUAL COSTS FACILITIES ANDADMINISTRATIVE COSTS 154,679 TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $ 423 687 PHS 398 (Rev. 04/06) Form Page 4 175

项目3：HDL代谢和反向胆固醇的分子调节 运输 高密度脂蛋白（HDL）及其主要蛋白载脂蛋白A-L（APOA-L）被认为可以保护 至少部分通过促进反向胆固醇转运至少部分地针对动脉粥样硬化心血管疾病 （RCT），从外围去除多余的胆固醇（例如血管壁）并返回 到肝脏排泄。该项目的广泛目标是对分子产生更多的了解 HDL代谢的生理学与反向胆固醇转运（RCT）有关 小鼠的模型并尽可能扩展实验。很明显 HDL胆固醇（HDL-C）的血浆浓度不足 而且，通过RCT途径和HDL功能的胆固醇通量的量度更为关键 关于对心血管疾病的影响。特定的目标1将涉及在体内使用鼠标 研究APOA-L的结构功能的实验，在补充物理学的研究中 项目2中的化学研究和项目1中的基于细胞的研究。ApoA-L变体将插入 基于AAV8的基因表达向量，将用于表达这些变体ApoA-L分子 APOA-L基因敲除小鼠以及对HDL代谢，RCT和选定情况的影响，动脉粥样硬化将为 决定。特定的目标2将使用鼠标模型，卵磷脂的作用：胆固醇的作用 酰基转移酶（LCAT），胆固醇酯转移蛋白（CETP）和磷脂转移蛋白（PLTP） 在调节巨噬细胞RCT的速率和与动脉粥样硬化的关系时。具体的目标3将 将小鼠发展的概念和方法扩展到人类环境，其特定目标的 使用新方法（ABCA1缺乏症）和LCAT测量RCT 不足。 虽然高水平的HDL胆固醇与冠状动脉疾病的风险降低有关 尚不清楚仅仅提高HDL胆固醇水平就足以降低心血管风险。这 HDL改变的机制以及对反向胆固醇转运过程的影响可能是 成为心血管风险的更重要的决定因素。该项目将寻求了解法规 活的老鼠和人类反向胆固醇的反向运输。 宾夕法尼亚大学医学院 翻译医学和治疗学研究所 654 BRBII/III 421 Curie Blvd 费城，宾夕法尼亚州19104 PHS 398（Rev. 04/06）表格2 173 首席研究员/计划主任（最后，第一，中间）：菲利普斯，迈克尔·C。 项目3 关键人员。请参阅说明。根据需要使用延续页面，以下面显示的格式提供所需的信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，首先姓氏。 姓名 Rader，Daniel J. 医学教授 病理学和药理学 Cuchel，码头 研究助理教授， 药品 其他重要的贡献者 姓名 Chaitanya Divgi 艾伦·雷梅利（Alan Remaley） ERA CONSONS用户名组织在项目中的角色 大学 PI的Danrader Pennsylvaniaschool 药品 大学 Billheij 大学 麦加尔·宾夕法尼亚州共同研究员学校 药品 组织角色 宾夕法尼亚大学合作者 NIH/NHLBI合作者 人类胚胎干细胞x否q是 如果所提出的项目涉及人类胚胎干细胞，请从以下列表中列出特定细胞系的注册数： http：//stemcells.nih.qov/reqistry/index.asp。根据需要使用延续页面。 如果目前无法引用特定行，请包括一个将使用注册表中的陈述。 细胞系 PHS 398（Rev. 04/06）表格第2页。 174 首席调查员 /计划主管（最后一个。中间）：菲利普斯。迈克尔·C· 项目3 从通过 初始预算期的详细预算 直接费用仅12/01/08 11/30/09 人员（仅申请人组织）致力于提高的月份 在Cal上的角色。学院。夏天 名称项目mnths mnths mnths 主要的 拉德，丹尼尔调查员2.40 Billheimer，Jeffrey共同投资者1.20 Cuchel，Marina共同入侵者0.6 博士后 Tanigawa，Hiroyuki 12.0 家伙 研究 Deborah Cromley 3.0 专家 研究 Aishawilson 6.0 专家 第2页 小计 顾问费用 设备（逐项） 供应（逐项） 塑料/玻璃器皿 化学物质，放射化合物和生物试剂 临床用品和研究药物 动物购买 旅行 患者护理费用住院 门诊 更改和翻新（按类别逐项列出） 其他费用（按类别逐项列出） 动物由Diems 邮政DOC健康保险 出版物 主题可观 财团/合同费用 要求的美元金额（省略美分） Inst.Base工资附带 薪金要求福利总计 **** 37,320 11,159 48,479 97,600 9,760 2,918 12,678 86,570 0 0 0 55,426 55,426 5,376 60,802 62,588 15,647 4,678 20,325 43,894 21,947 6,562 28,509 26,252 7,850 34,102 ^W 166,352 38,543 204,895 7,013 10,000 4,000 10,000 31,013 10,000 8,100 2,500 12.500 33,100 直接成本 初始预算期（项目7a。facePage）的小计直接费用$ 269 008 财团/合同费用设施和管理费用154,679 初始预算期的总直接成本$ 423 687 PHS 398（修订版04/06）表4页 175