Mitochondrial and Nuclear Mechanisms of Estrogen Receptor-Mediated Cardioprotecti

雌激素受体介导的心脏保护的线粒体和核机制

基本信息

批准号：
8230511
负责人：
DONNA HOPE KORZICK
金额：
$ 36.91万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-16 至 2014-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8230511
关键词：
1-Phosphatidylinositol 3-Kinase Acute Address Adult Age Aging Agonist American Amino Acid Sequence Animal Model Antioxidants Apoptosis Apoptotic Attenuated Binding Biochemical Biological Assay Cardiac Cardiovascular Diseases Cardiovascular system Cause of Death Cell Death Cell Survival Centrifugation Cessation of life Chronic Clinical Research Clinical Trials Computer software Conflict (Psychology)Coronary Data Demographic Aging Development Diagnosis Economic Burden Electron Spin Resonance Spectroscopy Enzymes Epidemiology Estrogen Nuclear Receptor Estrogen Receptors Estrogen Replacement Therapy Estrogens Event Female Gene Expression Genetic Transcription Genomics Glycogen Synthase Kinases Health Health Care Costs Heart Heart Diseases Heart failure Hospitals Image In Vitro Infarction Ischemia Label Ligation Mediating Mediator of activation protein Medical Membrane Mitochondria Mitochondrial Proteins Modeling Molecular Myocardial Infarction Necrosis Nitric Oxide Synthase Nuclear Nuclear Protein Ontology Operative Surgical Procedures Outcome Oxidative Stress Pathway interactions Peptide Sequence Determination Peptides Persons Phenotype Phosphorylation Phosphotransferases Physiological Population Post-Translational Protein Processing Postmenopause Production Protein Kinase Proteins Proteomics Rattus Receptor Activation Receptor Gene Receptor Signaling Reperfusion Injury Reperfusion Therapy Resources Risk Factors Role Scaffolding Protein Signal Pathway Signal Transduction Testing Therapeutic Time Up-Regulation Western Blotting Woman acute coronary syndrome age effect age related aged aging population cell growth regulation estrophilin improved in vivo juvenile animal men middle age mimetics mortality multiple reaction monitoring new therapeutic target non-genomic novel oxidant stress receptor expression research study senescence

项目摘要

DESCRIPTION (provided by applicant): Post-menopausal women are especially vulnerable to ischemic insult, and ~80,000 more cardiovascular deaths occur yearly in aged women than in aged men. Conflicting data exist on the role of estrogen (E2) in modulating ischemic tolerance (IT), due in part to the failed efficacy of E2 replacement to improve IT. Protective signaling pathways in the heart known to be influenced by E2 action include (PI3K)-Akt, eNOS and GSK3-2. Collectively, these signals impact mitochondrial events critical to cell survival to limit necrotic and apoptotic cell death. Here, we propose to test the novel paradigm that maladaptive estrogen receptor (ER) expression in the aged heart leads to a vicious cycle of increased oxidative stress (ROS) and ischemia/reperfusion (I/R injury), while selective engagement of non-genomic ER pathways can rescue this phenotype. We further propose that non-genomic ER signaling results in protein kinase C5 (PKC5) activation, a well-known mediator of cardioprotection. The following specific aims will be tested: 1) determine which ER subtype(s) reduce IT in aged, gonadectomized female rats, 2) determine the molecular mechanism(s) by which ERs alter ROS (ONOO- and O-2) through eNOS uncoupling, post-translational modification (PTM) or the PI3- Akt-GSK-32 axis, 3) determine the role of PKC5 in rapid ER signaling, and 4) identify novel mitochondrial PKC5 binding targets following non-genomic ER activation. Using a model of global ischemia, we will employ specific ER and PKC5 agonists and EPR spectroscopy to assess ROS. Apoptosis will be assessed through in vivo coronary ligation, western blotting for cellular ER subtype targeting and downstream signals, qRT- PCR for ER- mediated gene expression, and multiple reaction monitoring to assess PTMs of ERs. A targeted, state-of-the- art proteomic iTRAQ 8plex approach and LC MS/MS will be used to identify novel cardiac mitochondrial PKC5 protein partners and phosphorylation. We propose that non-genomic activation of ERs can compensate for E2 deficiency in aged female hearts, resulting in improved IT. The confirmation of our hypothesis will strongly support new targets for the treatment of acute coronary syndrome in the aging population and address the potential use of ER/PKC5 therapeutics as viable approaches to improve IT in post-menopausal women. PUBLIC HEALTH RELEVANCE: The leading cause of death among persons aged 75 years or older is heart disease, and deaths due to heart disease have actually increased by 60% from 1970 to 2002 among persons aged 80 years or older. Age is also the leading risk factor for the development of heart failure (CHF) and more Americans aged 65 and older are treated and discharged from hospitals with CHF than with any other diagnosis. Interestingly, mortality rates due to CHF are significantly greater in aged women vs aged men. CVD rates and associated mortality are also significantly greater in aged women when compared to aged men, and data from large scale clinical trials suggest that estrogen replacement therapy is ineffective in reducing, and may actually increase CVD outcomes. Collectively, this expanding aged demographic will undoubtedly require increased allocation of medical resources, raising the economic burden of health care costs for all Americans. The cellular mechanisms by which estrogen exerts effects in the aged heart, however, are poorly understood. The experiments associated with this proposal are expected to identify novel protein targets previously unstudied with estrogen deficiency. If our hypothesis on estrogen receptors is correct, new therapeutic targets for the treatment of ischemic cardiovascular disease in the aged population will be realized, i.e. use of estrogen receptor therapeutics as viable approaches to improve ischemic tolerance in aged women.

描述（由申请人提供）：绝经后女性特别容易受到缺血性损伤，每年老年女性因心血管死亡的人数比老年男性多约 80,000 例。关于雌激素 (E2) 在调节缺血耐受性 (IT) 方面的作用存在相互矛盾的数据，部分原因是 E2 替代疗法未能有效改善 IT。已知受 E2 作用影响的心脏保护性信号通路包括 (PI3K)-Akt、eNOS 和 GSK3-2。总的来说，这些信号影响对细胞生存至关重要的线粒体事件，以限制细胞坏死和凋亡。在这里，我们建议测试一种新的范例，即老年心脏中适应不良的雌激素受体（ER）表达导致氧化应激（ROS）增加和缺血/再灌注（I/R损伤）的恶性循环，同时选择性参与非基因组 ER 通路可以挽救这种表型。我们进一步提出非基因组 ER 信号传导导致蛋白激酶 C5 (PKC5) 激活，这是一种众所周知的心脏保护介质。将测试以下具体目标：1) 确定哪些 ER 亚型可减少老年、去腺术雌性大鼠的 IT，2) 确定 ER 通过 eNOS 改变 ROS（ONOO- 和 O-2）的分子机制解偶联、翻译后修饰 (PTM) 或 PI3-Akt-GSK-32 轴，3) 确定 PKC5 在快速 ER 信号传导中的作用，以及 4) 识别非基因组 ER 激活后的新线粒体 PKC5 结合靶点。使用全局缺血模型，我们将采用特定的 ER 和 PKC5 激动剂以及 EPR 光谱来评估 ROS。将通过体内冠状动脉结扎、用于细胞 ER 亚型靶向和下游信号的蛋白质印迹、用于 ER 介导的基因表达的 qRT-PCR 以及用于评估 ER 的 PTM 的多反应监测来评估细胞凋亡。最先进的靶向蛋白质组 iTRAQ 8plex 方法和 LC MS/MS 将用于鉴定新型心脏线粒体 PKC5 蛋白伴侣和磷酸化。我们认为 ER 的非基因组激活可以补偿老年女性心脏中 E2 的缺陷，从而改善 IT。我们假设的证实将有力地支持治疗老龄化人群中急性冠状动脉综合征的新目标，并解决 ER/PKC5 疗法作为改善绝经后妇女 IT 的可行方法的潜在用途。公共卫生相关性：75 岁或以上人群死亡的主要原因是心脏病，从 1970 年到 2002 年，80 岁或以上人群因心脏病死亡的人数实际上增加了 60%。年龄也是发生心力衰竭 (CHF) 的主要危险因素，65 岁及以上的美国人因 CHF 接受治疗并出院的人数多于其他任何疾病。有趣的是，老年女性因慢性心力衰竭导致的死亡率明显高于老年男性。与老年男性相比，老年女性的 CVD 发生率和相关死亡率也明显更高，大规模临床试验的数据表明，雌激素替代疗法无法有效减少 CVD 结果，实际上可能会增加 CVD 结果。总的来说，不断扩大的老龄化人口无疑需要增加医疗资源的分配，从而增加所有美国人医疗保健费用的经济负担。然而，人们对雌激素在衰老心脏中发挥作用的细胞机制知之甚少。与该提议相关的实验预计将鉴定出以前未研究过的雌激素缺乏的新蛋白质靶点。如果我们对雌激素受体的假设是正确的，那么治疗老年人群缺血性心血管疾病的新治疗目标将得以实现，即使用雌激素受体疗法作为改善老年女性缺血耐受性的可行方法。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

CALL FOR PAPERS Mitochondrial Function/Dysfunction in Health and Disease Local delivery of a PKCε-activating peptide limits ischemia reperfusion injury in the aged female rat heart

征文健康和疾病中的线粒体功能/功能障碍 PKCγ 激活肽的局部递送可限制老年雌性大鼠心脏的缺血再灌注损伤

DOI：
发表时间：
2024-09-13
期刊：
影响因子：
0
作者：
T. Lancaster;S. J. Jefferson;D. Korzick
通讯作者：
D. Korzick

心脏缺血再灌注损伤的年龄相关差异：雌激素缺乏的影响。

DOI：
发表时间：
2013-05
期刊：
影响因子：
0
作者：
Korzick, Donna H;Lancaster, Timothy S
通讯作者：
Lancaster, Timothy S

Age-dependent reductions in mitochondrial respiration are exacerbated by calcium in the female rat heart.

雌性大鼠心脏中的钙会加剧线粒体呼吸随年龄的减少。

DOI：
发表时间：
2012-06
期刊：
影响因子：
0
作者：
Hunter, J Craig;Machikas, Alexandra M;Korzick, Donna H
通讯作者：
Korzick, Donna H

Quantitative proteomic analysis reveals novel mitochondrial targets of estrogen deficiency in the aged female rat heart.

定量蛋白质组学分析揭示了老年雌性大鼠心脏中雌激素缺乏的新线粒体靶点。

DOI：
10.1152/physiolgenomics.00184.2011
发表时间：
2012-10-15
期刊：
Physiological genomics
影响因子：
4.6
作者：
T. Lancaster;Sarah J. Jefferson;J. Hunter;V. López;J. E. V. Eyk;E. Lakatta;D. Korzick
通讯作者：
D. Korzick

From syncitium to regulated pump: a cardiac muscle cellular update.

从合胞体到调节泵：心肌细胞更新。