Molecular Regulation of HDL Metabolism and Reverse Cholesterol Transport

项目3：高密度脂蛋白代谢和胆固醇逆转的分子调控 运输 高密度脂蛋白及其主要蛋白载脂蛋白A-L(载脂蛋白A-L)被认为可以保护 抗动脉粥样硬化性心血管疾病至少部分是通过促进胆固醇的反向转运 (RCT)，从而将多余的胆固醇从外周(如血管壁)移除并返回 进入肝脏排泄。这个项目的主要目标是加深对分子的理解。 利用体内整合技术研究高密度脂蛋白代谢与胆固醇反向转运的生理学关系 在老鼠身上建立模型，并在可能的情况下将实验扩展到人类。现在已经很明显了， 血浆高密度脂蛋白胆固醇浓度(高密度脂蛋白-C)不是抗动脉粥样硬化作用的适当替代品， 而通过RCT途径的胆固醇流量和高密度脂蛋白功能的测量对 关于对心血管疾病的影响。具体目标1将涉及在体内使用小鼠 通过实验研究载脂蛋白A-L的结构和功能特性，以补充其物理和功能特性。 项目2中的化学研究和项目1中的细胞研究。载脂蛋白A-L变体将插入 基于AAV8的基因表达载体，将用于表达这些变异的载脂蛋白A-L分子 载脂蛋白A-L基因敲除小鼠及其对高密度脂蛋白代谢、RCT的影响，在选定的病例中，动脉粥样硬化 下定决心。具体目标2将使用小鼠模型解决卵磷脂：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的比率及其与动脉粥样硬化形成的关系方面。具体目标3将 将在老鼠身上开发的概念和方法扩展到人类环境中，具体目标是 用新方法测量丹吉尔病(ABCA1缺乏症)和LCAT患者的RCT 缺乏症。 虽然高密度脂蛋白水平与降低冠状动脉疾病的风险有关，但它是 尚不清楚仅仅提高高密度脂蛋白水平就足以降低心血管风险。这个 高密度脂蛋白改变的机制及其对胆固醇反向转运过程的影响 成为心血管风险的一个更重要的决定因素。本项目将寻求了解该规则 活着的小鼠和人类体内胆固醇的反向运输。 宾夕法尼亚大学医学院 转化医学和治疗学研究所 654 BRBII/III 居里大道421号 宾夕法尼亚州费城，邮编19104 小灵通398(04/06版)表格第2页 173 首席研究员/项目主任(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。根据需要使用续页，以如下所示的格式提供所需信息。 从首席调查员(S)开始。按字母顺序列出所有其他关键人员，姓氏在前。 名字 丹尼尔.雷德，Daniel J. 医学教授， 病理学与药理学 库切尔，马里纳 研究助理教授 医学 其他重要贡献者 名字 柴坦尼亚·迪夫吉 阿兰·雷马利 Era Commons用户名组织在项目中的角色 伊利诺伊大学 宾夕法尼亚丹瑞德猪学院 医学 中华人民共和国大学 BILLHIJ 伊利诺伊大学 宾夕法尼亚大学联合调查员学院 医学 项目中的组织角色 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 如果建议的项目涉及人类胚胎干细胞，请从下面的列表中列出特定细胞系(S)的注册号： Http://Stemcells.nih.qov/reqistry/index.asp.根据需要使用续页。 如果此时不能引用某一特定行，请附上一项声明，说明将使用登记处的某一行。 细胞系 小灵通398(04/06版)表格第2页-续 一百七十四 首席调查员/项目主任(最后第一。中间)：菲利普斯。迈克尔·C。 项目3 从头到尾 初步预算期明细预算 直接成本仅12/01/08 11/30/09 人员(仅限申请组织)致力于项目的月份 在Cal上的角色。阿卡德。夏天 项目名称mnths mnths mnths 本金 雷德，丹尼尔调查员2.40 比尔海默，杰弗里联合调查员1.20 Cuchel，Marina共同调查员0.6 博士后 谷川广之12.0 同胞 研究 Deborah Cromley 3.0 专家 研究 艾莎·威尔逊6.0 专家 第2页 小计 顾问费 设备(分项) 供应品(分项) 塑料/玻璃器皿 化学品、放射性化学品和生物试剂 临床用品和研究用药 动物采购 旅行 住院病人护理费用 门诊病人 改建和翻新(按类别分列) 其他费用(按类别分项列出) 按日计的动物 博士后医疗保险 出版物 受试者补偿 联合体/合同成本 请求的美元金额(省略美分) INST.BASE工资福利 薪资申请福利合计 *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 四千 10,000 31,013 10,000 8,100 2500人 12.500 33,100 直接成本 初步预算期的直接费用小计(项目7a。面页)$269 008 联合体/合同费用设施和行政费用154,679 初步预算期间的直接费用总额423 687美元 小灵通398(04/06版)表格第4页 175

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