Mechanisms for Altered Glucose Homeostasis During HAART

HAART 期间改变血糖稳态的机制

基本信息

批准号：
7764755
负责人：
PAUL W HRUZ
金额：
$ 29.49万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-04-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7764755
关键词：
Acute Adipocytes Adipose tissue Animals Antibodies Antibody Affinity Atazanavir Binding Binding Proteins Binding Sites Cardiac Chronic Co-Immunoprecipitations Continuous Intravenous Infusion Control Animal Coupled Deoxyglucose Development Diabetes Mellitus Epitopes Euglycemic Clamping Fatty acid glycerol esters Funding GLUT 4 protein GLUT4 gene Glucose Clamp Glucose Transporter Goals HIV Protease Inhibitors Heart Hepatic Highly Active Antiretroviral Therapy Indinavir Individual Insulin Insulin Resistance Knockout Mice Label Leptin Mass Spectrum Analysis Measures Mediating Metabolic Metabolic syndrome Mitochondria Molecular Molecular Target Morbidity - disease rate Muscle Mutation Nonesterified Fatty Acids Nucleosides Patients Peptides Peripheral Pharmaceutical Preparations Post-Translational Protein Processing Production Proteins RNA Radiolabeled Rattus Retinol Binding Proteins Reverse Transcriptase Inhibitors Ritonavir Role SLC2A1 gene Serum Skeletal Muscle Stavudine System Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Western Blotting Work Xenopus oocyte Zidovudine abacavir adiponectin basal insulin blood glucose regulation functional loss glucose disposal glucose production glucose transport glucose uptake intraperitoneal mortality prevent radiotracer research study resistin uptake

项目摘要

DESCRIPTION (provided by applicant): The long term goal of this project is to understand the molecular mechanisms responsible for altered glucose homeostasis during highly active antiretroviral therapy (HAART). The metabolic changes that occur in association with the use of HIV protease inhibitors (Pis) have significant potential for increasing cardiac- associated morbidity and mortality. The immediate objectives of this proposal are to determine the specific contribution of GLUT4 inhibition to altered glucose uptake into insulin responsive tissues, to elucidate the role of altered glucose transport in the development of changes in adiocytokine production, and to identify the precise molecular interactions that mediate PI binding to GLUT4. The role of GLUT4 in Pi-induced effects on peripheral glucose disposal will be studied by measuring radiolabeled 2-deoxyglucose uptake into cardiac, skeletal muscle and adipose tissue under basal and hyperinsulinemic euglycemic clamp conditions in wild-type and transgenic mice with specific deletion of GLUT4 from muscle or fat. Changes in leptin, adiponectin and resistin RNA and protein levels, peripheral glucose disposal, and hepatic glucose production will be determined following semi-acute exposure of control and GLUT4 null mice to Pis in the presence and absence of semi-chronic treatment with NRTIs. The structural features in GLUT4 that mediate HIV protease inhibitor binding and inactivation will be determined in primary and/or cultured rat adipocytes labeled with a photoactivatable peptide containing the GLUT4 inhibiting epitope zHFF coupled to the FLAG epitope. Modified proteins will be isolated using a FLAG antibody affinity column and identified by mass spectrometry. The ability of Pis to influence the binding of proteins known to interact with the GLUT4 carboxy terminus will also be investigated by Western blot analysis and co-immunoprecipitation experiments using factor specific antibodies. Mutations will be introduced into the identified GLUT4 PI binding site and the resulting functional effects on transport activity and PI sensitivity will be determined. Taken together, these studies will provide crucial information about the mechanism(s) by which Pis contribute to the development of insulin resistance and diabetes mellitus. This will greatly assist efforts to develop effective strategies for preventing and/or treating the metabolic changes that occur both in patients on HAART and in the wider context of HIV-negative individuals with features of the metabolic syndrome.

描述（由申请人提供）：该项目的长期目标是了解高度活跃抗逆转录病毒疗法（HAART）期间导致葡萄糖稳态改变的分子机制。与使用HIV蛋白酶抑制剂（PI）相关的代谢变化具有增加心脏相关的发病率和死亡率的显着潜力。该提案的直接目的是确定GLUT4抑制对改变葡萄糖摄取对胰岛素反应组织的特定贡献，以阐明葡萄糖转运改变在adocytokine产生变化的变化中的作用，并确定介导PI与Glut4结合的精确分子相互作用。通过测量在野生型和特定的glut4 glut4 fat 4的基础和高胰岛素的糖脂条件下，将研究GLUT4在PI诱导的对周围葡萄糖处置作用中的作用，以测量放射性标记的2-脱氧葡萄糖吸收在基础和高胰岛素血液远产血夹中的心脏，骨骼肌和脂肪组织中。瘦素，脂联素和抗蛋白RNA以及蛋白质水平，外周葡萄糖处置以及肝葡萄糖产生的变化将在半急性对照和GLUT4 NULL小鼠的半急性暴露后，在存在和不使用NRTI的半偶然处理的情况下对PIS确定。 GLUT4中介导HIV蛋白酶抑制剂结合和失活的结构特征将在用含有lut4抑制表位ZHFF的光活性肽标记的原代和/或培养的大鼠脂肪细胞中确定，该肽偶联到标志表位。修饰的蛋白质将使用FLAG抗体亲和力柱分离，并通过质谱鉴定。也将通过使用特定于因子特定抗体的蛋白质印迹分析和共免疫沉淀实验来研究PI影响已知与GLUT4羧基末端相互作用的蛋白质结合的能力。突变将引入已鉴定的GLUT4 PI结合位点，并确定所得的功能对运输活性的影响，并确定PI敏感性。综上所述，这些研究将提供有关PI有助于胰岛素抵抗和糖尿病的发展机制的关键信息。这将极大地有助于制定有效的策略，以防止和/或治疗HAART患者以及具有代谢综合征特征的HIV阴性患者的代谢变化。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

HIV protease inhibitors act as competitive inhibitors of the cytoplasmic glucose binding site of GLUTs with differing affinities for GLUT1 and GLUT4.

DOI：
10.1371/journal.pone.0025237
发表时间：
2011
期刊：
PloS one
影响因子：
3.7
作者：
Hresko RC;Hruz PW
通讯作者：
Hruz PW

Molecular mechanisms for insulin resistance in treated HIV-infection.

DOI：
10.1016/j.beem.2010.10.017
发表时间：
2011-06
期刊：
Best practice & research. Clinical endocrinology & metabolism
影响因子：
0
作者：
Hruz PW
通讯作者：
Hruz PW

GS-8374, a novel HIV protease inhibitor, does not alter glucose homeostasis in cultured adipocytes or in a healthy-rodent model system.

GS-8374 是一种新型 HIV 蛋白酶抑制剂，不会改变培养脂肪细胞或健康啮齿动物模型系统中的葡萄糖稳态。

DOI：
10.1128/aac.01184-10
发表时间：
2011
期刊：
Antimicrobial agents and chemotherapy
影响因子：
4.9
作者：
Hruz,PaulW;Yan,Qingyun;Tsai,Luong;Koster,Joseph;Xu,Lianhong;Cihlar,Tomas;Callebaut,Christian
通讯作者：
Callebaut,Christian