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

• 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
• 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

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侮辱，老年女性每年发生的心血管死亡人数比老年男性多8万人。关于雌激素(E2)在调节缺血耐受(IT)中的作用，目前存在相互矛盾的数据，部分原因是E2替代治疗改善IT的效果失败。已知心脏中受E2作用影响的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存活至关重要的线粒体事件，以限制坏死和凋亡的细胞死亡。在这里，我们建议测试一种新的范式，即老年心脏中不适应性雌激素受体(ER)的表达导致氧化应激(ROS)增加和缺血/再灌注(I/R损伤)的恶性循环，而选择性地参与非基因组ER通路可以挽救这一表型。我们进一步提出，非基因组ER信号导致蛋白激酶C5(PKC5)激活，PKC5是一种众所周知的心脏保护介质。将测试下列特定目标：1)确定哪种ER亚型(S)可降低老年去性腺大鼠的IT；2)确定ER通过eNOS解偶联、翻译后修饰(PTM)或PI3-Akt-GSK-32轴改变ROS(ONOO-和O-2)的分子机制(S)；3)确定PKC5在快速ER信号转导中的作用；以及4)确定非基因组ER激活后线粒体PKC5新的结合靶点。利用全脑缺血模型，我们将使用特定的ER和PKC5激动剂和EPR波谱来评估ROS。将通过体内冠脉结扎、Western blotting检测细胞ER亚型靶向和下游信号、qRT-PCR检测ER介导的基因表达以及多反应监测来评估ER的PTM，以评估细胞凋亡。一种有针对性的、最先进的蛋白质组学iTRAQ 8plex方法和LC MS/MS将被用于鉴定新的心肌线粒体PKC5蛋白伙伴和磷酸化。我们认为，ER的非基因组激活可以弥补老年女性心脏中E2的缺乏，从而改善IT。我们的假设的证实将有力地支持老年人口中治疗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岁或以上人口的主要死因是心脏病，从1970年到2002年，80岁或以上人口中死于心脏病的人数实际上增加了60%。年龄也是心力衰竭(CHF)发展的主要风险因素，65岁及以上的美国人接受CHF治疗并出院的人数比其他任何诊断都要多。有趣的是，老年女性的CHF死亡率明显高于老年男性。与老年男性相比，老年女性的心血管疾病发病率和相关死亡率也明显更高，来自大规模临床试验的数据表明，雌激素替代疗法在减少心血管疾病方面无效，实际上可能会增加心血管疾病的结局。总体而言，不断扩大的老年人口无疑将需要增加医疗资源的分配，从而增加所有美国人的医疗费用的经济负担。然而，雌激素在老年心脏中发挥作用的细胞机制却知之甚少。与这一提议相关的实验有望确定以前未被研究过的雌激素缺乏的新蛋白质靶点。如果我们关于雌激素受体的假设是正确的，那么在老年人群中治疗缺血性心血管疾病的新的治疗目标将被实现，即使用雌激素受体治疗作为提高老年女性缺血耐受性的可行方法。

项目成果

Acute adiponectin delivery is cardioprotective in the aged female rat heart.

• DOI: 10.1111/ggi.12306
• 发表时间: 2015-05
• 期刊: Geriatrics & gerontology international
• 影响因子: 3.3
• 作者: Tomicek NJ;Hunter JC;Machikas AM;Lopez V;Korzick DH
• 通讯作者: Korzick DH

Age-related differences in cardiac ischemia-reperfusion injury: effects of estrogen deficiency.

• DOI: 10.1007/s00424-013-1255-7
• 发表时间: 2013-05
• 期刊: PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY
• 影响因子: 4.5
• 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.

• DOI: 10.1016/j.genm.2012.04.001
• 发表时间: 2012-06
• 期刊: Gender medicine
• 作者: Hunter JC;Machikas AM;Korzick DH
• 通讯作者: Korzick DH

Increased estrogen receptor β in adipose tissue is associated with increased intracellular and reduced circulating adiponectin protein levels in aged female rats.

脂肪组织中雌激素受体β 的增加与老年雌性大鼠细胞内脂联素蛋白水平的增加和循环脂联素蛋白水平的降低有关。

• DOI: 10.1016/j.genm.2011.05.010
• 发表时间: 2011
• 期刊: Gender medicine. official journal of the Partnership for Gender-Specific Medicine at Columbia University
• 作者: Tomicek,NanetteJ;Lancaster,TimothyS;Korzick,DonnaH
• 通讯作者: Korzick,DonnaH

Estrogen receptor beta does not influence ischemic tolerance in the aged female rat heart.

雌激素受体β不影响老年雌性大鼠心脏的缺血耐受性。

• DOI: 10.1111/j.1755-5922.2011.00288.x
• 发表时间: 2013
• 期刊: Cardiovascular therapeutics
• 影响因子: 3.1
• 作者: Tomicek,NanetteJ;Miller-Lee,JenniferL;Hunter,JCraig;Korzick,DonnaH
• 通讯作者: Korzick,DonnaH