Modulation of Endogenous PPAR Activation

对于三种不同的内源性途径，可能会产生负面影响 调节PPAR活性。这些机制将通过研究它们对已建立的 PPAR激活的体外和体内模型。 LPL介导的PPARa激活提示不同结构的脂肪酸对内皮的影响 可能是由于不同的PPARa激活，包括拮抗。在Aim 1中，PPAR的激活和抑制 通过特定的脂肪酸将在体外和体内进行研究，将omega-3脂肪酸与饱和和 反式脂肪酸。肝脏核因子4α(HNF4a)是一种知之甚少但却至关重要的脂肪酸。 激活的受体，调节脂肪代谢、血栓形成和血糖控制。通过使用 在我们的LPL/PPARa研究中，我们发现了新的脂代谢操作 HNF4a的调控机制及PPARa和HNF4a对HNF4a信号通路的分化反应 脂类代谢。在目标2中，将探索HNF4a和PPARa之间的这种差异。一本小说但是 直接内源性PPAR拮抗剂的存在可能是PPAR调控的重要机制。 虽然β胡萝卜素的对称切割产生了RXR核受体的天然配体，但我们有 发现不对称的β-胡萝卜素裂解会产生一种特定的载脂胡萝卜素，直接对抗 PPAR响应。在目标3中，这种直接拮抗剂将在体外和体内进行表征。加在一起，这些 研究结合了生化、生物和体内模型，以更好地了解内源性调节如何 PPAR活性的高低可能决定生物反应。 主要人员： 名字 普卢茨基，马里兰州豪尔赫 加齐亚诺，J.迈克尔，医学博士，公共卫生硕士 奥拉萨努，加布里埃拉，马里兰州 雷德，丹尼尔，马里兰州 肖尔森，斯蒂芬，医学博士 Ziouzenkova，Ouliana，博士 组织 布里格姆妇女医院 布里格姆妇女医院 布里格姆妇女医院 宾夕法尼亚大学 乔斯林糖尿病中心 布里格姆妇女医院 在项目中的角色 首席调查员 协作者 博士后研究员 协作者 协作者 研究助理 PHS 398/2590(05/01版)第154页延续格式Paqe 首席调查员/项目主任(最后，第一中)：米歇尔。托马斯 从头到尾 初步预算期详细预算4/1/05 3/31/06 仅直接成本 人员(仅适用于申请组织)申请金额百分比(省略美分) 键入Effort Inst. 在APPT中的角色。关于基本工资和福利 项目名称(月)项目。薪资申请福利合计 普鲁茨基校长12 20 170,000 34,000 10,880 44,880 调查员 Ouliana Ziouzenkova Res.12 50 50 50 25 000 8 000 33 000 阿索克。 技术员12 50 28,000 14,000 4,060 18,060 Gabriela Orasanu Post-Doc 12 100 30 30 30 6 300 36 300 同胞 技术员12 100 28,000 28,000 8,120 36,120 131,000 37,360 168,360 超级表 顾问费 “RJ‘”“ 设备(分项) 无 ‘&lt；*？&lt；’r？“‘ ？-.&lt； 用品(按类别分项列出) ？* 分子生物学。试剂(酶、转染剂、核受体分析)：11200 ，‘-“&> 免疫试剂(抗体、免疫试剂等)：8,014？-r？ &lt；Kij“i TBisoscuhemCiuclatulsre(A(Mpoe1d4ia，syPnCthSe，Spisla，sPtiPcwararae/)P：P10A，R78g0激动剂，放射性同位素)：10,500 .K\“ *40,494“ 旅行‘I！1*！&gt；，*ffi’‘，，-&gt；！’！&gt；？‘，IV 病人护理费用住院病人0 门诊部0 改建和翻新(按类别分列) “0 其他费用(按类别分项)：“L”\J&>；-~~“？”-V- T.-：我 鼠标板，总计，1年：9412美元 动物设置：5笼？3.88美元/笼=19美元 动物设施费用：4.715美元[50%(食宿+食宿+套餐) 14,146 初期预算期间直接费用小计223 000美元 财团/直接成本n 合同成本、设施和管理成本0 初步预算期的直接费用总额(项目7a，正面)223 000美元 小灵通398(05/01版)页面表格第4页 一百五十五

for three different endogenous pathways that may negatively regulate PPAR activity. These mechanisms will be studied by examining their modulation of well-established in vitro and in vivo models of PPAR activation. LPL-mediated PPARa activation suggests the discrepant endothelial effects of structurally diverse fatty acids may be due to differential PPARa activation, including antagonism. In Aim 1, PPAR activation and inhibition by specific fatty acids will be studied in vitro and in vivo, contrasting omega-3 fatty acids to saturated and trans-fatty acids. Hepatic nuclear factor 4 alpha (HNF4a) is a poorly understood but critical fatty acid- activated receptor that regulates lipid metabolism, thrombosis, and glucose control. By using the manipulations of lipid metabolism employed in our LPL/PPARa studies, we have identified novel mechanisms of HNF4a modulation and divergent responses between PPARa and HNF4a to pathways of lipid metabolism. In Aim 2, this divergence between HNF4a and PPARa will be explored. A novel but powerful mechanism for PPAR modulation would be the existence of a direct endogenous PPAR antagonist. While symmetric cleavage of beta carotene generates natural ligands for the RXR nuclear receptor, we have identified that asymmetric beta carotene cleavage produces a specific apocarotenal that directly antagonizes PPAR responses. In Aim 3, this direct antagonist will be characterized in vitro and in vivo. Together, these studies integrate biochemical, biologic, and in vivo models to better understand how endogenous modulation of PPAR activity may determine biologic responses. KEY PERSONNEL: Name Plutzky, Jorge, MD Gaziano, J. Michael, MD, MPH Orasanu, Gabriella, MD Rader, Daniel, MD Shoelson, Stephen, MD, PhD Ziouzenkova, Ouliana, PhD Organization Brigham and Women's Hospital Brigham and Women's Hospital Brigham and Women's Hospital University of Pennsylvania Joslin Diabetes Center Brigham and Women's Hospital Role on Project Prinicipal Investigator Collaborator Post-doctoral Fellow Collaborator Collaborator Research Associate PHS 398/2590 (Rev. 05/01) Page 154 Continuation Format Paqe Principal Investigator/Program Director (Last, first. Middle): Michel. Thomas FROM THROUGH DETAILED BUDGET FOR INITIAL BUDGET PERIOD 4/1/05 3/31/06 DIRECT COSTS ONLY PERSONNEL (Applicant organization only) % DOLLAR AMOUNT REQUESTED (omit cents) TYPE EFFORT INST. ROLE ON APPT. ON BASE SALARY FRINGE NAME PROJECT (MONTHS) PROJ. SALARY REQUESTED BENEFITS TOTALS Jorge Plutzky Principal 12 20 170,000 34,000 10,880 44,880 Investigator Ouliana Ziouzenkova Res. 12 50 50,000 25,000 8,000 33,000 Assoc. Technician Tech 12 50 28,000 14,000 4,060 18,060 Gabriela Orasanu Post-Doc 12 100 30,000 30,000 6,300 36,300 Fellow Technician Tech 12 100 28,000 28,000 8,120 36,120 131,000 37,360 168,360 SUEJTOTALS CONSULTANT COSTS " "" rj '"" EQUIPMENT (Itemize) None ' <* ¿< ' r ¿" ' ¿?- .< SUPPLIES (Itemize by category) ¿ * Molecular Bio. Reagents (Enzymes, transfection reagents, nuclear receptor assays): 1 1 ,200 ,'-">¿ - Immunoreagents (Antibodies, Western reagents, etc): 8,014 ¿ ~¿. -r¿ <K ij" i TBisoscuhemCiuclatulsre(A(Mpoe1d4ia,syPnCthSe,spisla,sPtiPcwARarae/)P: P10A,R78g0agonists, radioisotope): 10,500 .-'if ¿ .K \" *40,494 " TRAVEL 'I!1*! > , *ffi'',, ->!'!>? ', IV PATIENT CARE COSTS INPATIENT 0 OUTPATIENT 0 ALTERATIONS AND RENOVATIONS (Itemize by category) " 0 OTHER EXPENSES (Itemize by category) ffslt ' l' \ J>-~~ '¿" -V- t .-:i Mouse Board, total, Yr 1 :$9412 Animal set-up: 5 cages¿ $3.88/cage = $19 Animal Facility Fee: $4.715 [50%(rjurchase+board+set-uDll 14,146 SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $223,000 CONSORTIUM/ DIRECT COSTS n CONTRACTUAL COSTS FACILITIES AND ADMINISTRATION COSTS 0 TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD (Item 7a,Face Page) $223,000 PHS 398 (Rev. 05/01) Page Form Page 4 155