Role of Glutathione S-Transferase P in Heart Failure

谷胱甘肽 S-转移酶 P 在心力衰竭中的作用

基本信息

批准号：
8013061
负责人：
Sumanth D Prabhu
金额：
$ 37.25万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2011-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8013061
关键词：
4 hydroxynonenal Ablation Accounting Acrolein Affect Aldehydes Alleles Angiogenic Proteins Anti-Inflammatory Agents Anti-inflammatory Antioxidants Apoptosis Biological Markers Bone Marrow Cardiac Cardiomyopathies Cell physiology Chronic Clinical Clinical Trials Coronary Depressed mood Diagnostic Disease Disease susceptibility Drug Metabolic Detoxication Enzymes Failure Fibrosis Functional disorder GSTP1 gene Gene Expression Generations Genetic Genetic Polymorphism Glutathione Glutathione S-Transferase Glutathione S-Transferase P Goals Heart Heart failure Hematopoietic Human Hydrogen Peroxide Individual Differences Infarction Inflammation Inflammatory Injury Isotope Labeling Left Ventricular Remodeling Ligation Lipid Peroxidation Lipid Peroxides Malignant Neoplasms Mass Spectrum Analysis Measures Membrane Metabolic Metabolism Modeling Mus Muscle Cells Myeloproliferation Myocardial Myocardium N-terminal Oxidants Oxidation-Reduction Oxidative Stress Patients Peroxidases Peroxides Phosphotransferases Play Predisposition Process Property Protein Isoforms Proteins Reactive Oxygen Species Role Selenium Stem cells Stress Systemic disease Testing Therapeutic Tissues Transgenic Mice Transgenic Organisms Variant Wild Type Mouse Work Xenobiotics adduct antioxidant therapy base biological adaptation to stress chemotherapy cytotoxic design gain of function in vivo neovascularization novel overexpression peroxiredoxin public health relevance regenerative therapy repaired response stress-activated protein kinase 1

项目摘要

DESCRIPTION (provided by applicant): Although oxidative stress is a hallmark of heart failure (HF), clinical trials with antioxidants targeting first generation reactive oxygen species (ROS) such as O2/ and H2O2 have not yielded compelling benefits. However, ROS also generate secondary intermediates derived from lipid peroxidation, including peroxides and aldehydes that amplify oxidative injury. Glutathione S-transferases (GSTs) metabolize aldehydes by catalyzing their conjugation with glutathione (GSH). Select GST isoforms also have important non-catalytic functions such as physical interactions with c-Jun N-terminal kinase (JNK) that are modulated by oxidative stress. Although GSTs play a vital role in oxidative stress responses, how GSTs impact HF is unknown. Our goal is to define the functional role of GSTP, most abundant cardiac GST isoform, in HF. Our preliminary studies indicate that GSTP is downregulated in HF, and that GSTP deficiency worsens cardiac remodeling, augments protein- aldehyde adducts, depresses circulating endothelial progenitor cells (EPCs), and impairs neovascularization. Our central hypothesis, therefore, is that GSTP is a critical cardioprotective protein in post-infarction HF that ameliorates remodeling and promotes cardiac repair. To test this hypothesis, we will perform three Specific Aims. In Aim 1, we will define the role of GSTP, and the human GSTP variants hGSTP1*A and hGSTP1*C, in HF by examining post-infarction LV remodeling in wild-type (WT), GSTP-/-, and cardiac-specific hGSTP1*A and hGSTP1*C transgenic (Tg) mice. We will evaluate apoptosis, fibrosis, and inflammation together with glutathione levels, protein-adducted aldehydes, and JNK activation in the heart. In Aim 2, we will determine the metabolic contribution of GSTP to the detoxification of lipid peroxidation products in the failing heart. In isolated, perfused sham-operated and failing hearts from WT, GSTP-/- and hGSTP Tg mice, using isotope labeling and mass spectrometry, we will characterize the metabolism and detoxification of unsaturated aldehydes. In tissue homogenates, we will also determine GSTP-related peroxidase activity and levels of aldehydes and lipid peroxides. In Aim 3, we will delineate the cardiac and bone marrow (BM)-related effects by which GTSP modulates neovascularization in the failing heart. We will first determine EPC and BM progenitor cell function in WT and GSTP -/- mice, both with and without concomitant JNK inhibition. Next, we will define how GSTP ablation and hGSTP overexpression affect neovascularization and angiogenic gene expression in the sham and failing hearts from Aim 1. Lastly, we will evaluate post-infarction remodeling, inflammation, EPC mobilization, and neovascularization in chimeric mice: WT mice with GSTP-/- BM and GSTP-/- mice with WT BM. These studies will establish the role of myocardium-localized versus BM-localized GSTP in the process of remodeling, neovascularization, and inflammation in the failing heart. Collectively, this work will establish a novel paradigm of GSTP as an essential antioxidant, anti-inflammatory, and pro-angiogenic protein in HF. This model can have important diagnostic and therapeutic implications for HF patients with regard to oxidant injury. PUBLIC HEALTH RELEVANCE: These studies will establish glutathione S-transferase P (GSTP) as a critical antioxidant and tissue reparative protein in heart failure and identify new determinants of oxidative injury. Hence, the results can help design new, non-classical antioxidant and regenerative therapies in heart failure. We will also evaluate the cardioprotective potency of human GSTP variants in the failing heart; important differences between GSTP variants can establish a genetic basis for variability of antioxidant responses in the heart and provide a novel biomarker for the progression of heart failure.

描述（由申请人提供）：尽管氧化应激是心力衰竭的标志（HF），但针对第一代活性氧（ROS）（例如O2/和H2O2）的抗氧化剂的临床试验尚未带来令人信服的好处。然而，ROS还产生从脂质过氧化的次级中间体，包括扩增氧化损伤的过氧化物和醛。谷胱甘肽S-转移酶（GSTS）通过催化与谷胱甘肽（GSH）的结合来代谢醛。选择的GST同工型也具有重要的非催化功能，例如与氧化应激调节的C-Jun N末端激酶（JNK）的物理相互作用。尽管GST在氧化应激反应中起着至关重要的作用，但GST如何影响HF是未知的。我们的目标是在HF中定义GSTP，最丰富的心脏GST同工型的功能作用。我们的初步研究表明，GSTP在HF中被下调，而GSTP缺乏会使心脏重塑恶化，增加蛋白质 - 醛加合物，降低循环的内皮祖细胞（EPC），并损害异常性。因此，我们的中心假设是，GSTP是炎后HF中的关键心脏保护蛋白，可改善重塑并促进心脏修复。为了检验这一假设，我们将执行三个具体目标。在AIM 1中，我们将通过检查野生型（WT），GSTP - / - 和CardIAC-和CardIAC-和CardIAC-和CardIAC-和Cardiac-，在HF中定义了人GSTP的作用HGSTP1*A和HGSTP1*c，HGSTP1*A和HGSTP1*C。特定的HGSTP1*A和HGSTP1*C转基因（TG）小鼠。我们将在心脏中评估凋亡，纤维化和炎症，以及谷胱甘肽水平，蛋白质的醛和JNK激活。在AIM 2中，我们将确定GSTP对失败心脏中脂质过氧化产物排毒的代谢贡献。在使用同位素标记和质谱法的孤立的，灌注的假手术和失败的心脏中，我们将表征不饱和醛的代谢和解毒。在组织匀浆中，我们还将确定与GSTP相关的过氧化物酶活性以及醛和脂质过氧化物的水平。在AIM 3中，我们将描绘出与GTSP调节失败心脏的新生血管形成的心脏和骨髓（BM）相关效应。我们将首先确定WT和GSTP - / - 小鼠中的EPC和BM祖细胞功能，无论有没有伴随的JNK抑制作用。接下来，我们将定义GSTP消融和HGSTP的过表达如何影响假手术中的新血管形成和血管生成基因表达，而心脏的失败心脏1。最后，我们将评估输入后的重塑，炎症，EPC动员，EPC动员和新生血管中的新虫小鼠：WT小鼠：WT小鼠：WT小鼠：WT小鼠：WT小鼠：WT小鼠：WT小鼠：使用GSTP - / - BM和GSTP - / - 带有WT BM的小鼠。这些研究将确定心肌定位与BM定位的GSTP在失败的心脏中重塑，新生血管形成和炎症的过程中的作用。总的来说，这项工作将建立一种新型的GSTP范式作为HF中必不可少的抗氧化剂，抗炎和促血管生成蛋白。对于氧化剂损伤，该模型对HF患者具有重要的诊断和治疗意义。公共卫生相关性：这些研究将建立谷胱甘肽S-转移酶P（GSTP）作为心力衰竭中的一种关键抗氧化剂和组织修复蛋白，并确定氧化损伤的新决定因素。因此，结果可以帮助设计心力衰竭的新的，非经典的抗氧化剂和再生疗法。我们还将评估失败心脏中人类GSTP变体的心脏保护效力； GSTP变体之间的重要差异可以为心脏中抗氧化反应的变异性建立遗传基础，并为心力衰竭发展提供新颖的生物标志物。