Mechanisms of Heterocellular Signaling at the Myoendothelial Junction

肌内皮连接处的异细胞信号传导机制

• 批准号: 9249957
• 负责人: Brant E Isakson
• 金额: $ 52.04万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9249957
• 关键词: Ablation; Acetylcholine; Alleles; Anatomy; Animal Model; Animals; Arteries; Binding; Biochemical Genetics; Blood; Blood Pressure; Blood Vessels; Blood flow; Bone Marrow Transplantation; Cells; Coculture Techniques; Computer Simulation; Consumption; Data; Derivation procedure; Diffusion; Dilator; Dioxygenases; Disease model; Endothelial Cells; Endothelium; Erythrocytes; Genes; Heme Iron; Hemoglobin; Hemoglobin A; Hemoglobin Chaperone Pathway; Human; Hypertension; Knock-in Mouse; Knockout Mice; Link; Location; LoxP-flanked allele; Macromolecular Complexes; Mediating; Medical; Modeling; Molecular Chaperones; Molecular Models; Mouse Protein; Mouse Strains; Mus; Mutation; NOS3 gene; Nitric Oxide; Output; Oxidation-Reduction; Oxygen; Pathologic; Perfusion; Peripheral; Peripheral Resistance; Pharmaceutical Preparations; Pharmacology; Phenylephrine; Physiological; Physiology; Play; Point Mutation; Protein Overexpression; Proteins; Pump; Recombinant Proteins; Regional Blood Flow; Regulation; Resistance; Role; Signal Transduction; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myocytes; Source; Stimulus; Structure; Sum; Testing; Time; Tissues; Vascular resistance; Work; alpha-Thalassemia; base; blood lead; blood pressure reduction; blood pressure regulation; cadherin 5; constriction; cytochrome b5 reductase; experimental study; feeding; genetic approach; genetic manipulation; genome-wide; glycosylated-nitric oxide complex hemoglobin A; hypertension control; improved; insight; knock-down; molecular modeling; mutant; novel; overexpression; peripheral blood; protein expression; public health relevance; response; small hairpin RNA; vascular bed

DESCRIPTION (provided by applicant): We have recently discovered that hemoglobin � is enriched in myoendothelial junctions, the anatomical location where endothelial cells and smooth muscle cells make contact in the resistance arteries. This was a significant finding because it demonstrated that hemoglobin � had an important and active role outside of erythrocytes. This protein is one of only a few truly polarized proteins to be localized to endothelial-derived myoendothelial junctions, and the siRNA-induced decrease in the amount of the protein significantly altered arterial reactivity, including constriction to phenylephrine and dilation to acetylcholine. The mechanism we derived was based on evidence indicating that monomeric hemoglobin � is a potent scavenger of nitric oxide, and that endothelial nitric oxide synthase (eNOS) and hemoglobin � were found to be in a macromolecular complex. Based on this work, as well as a plethora of strong preliminary data, we hypothesize that hemoglobin � at the myoendothelial junction is a novel regulator of nitric oxide signaling which can impact blood pressure regulation. We will test this hypothesis using two specific aims: 1.) investigate the effects of endothelial hemoglobin a gene ablation/over-expression on arterial function and 2.) elucidate how AHSP and eNOS regulate hemoglobin � expression and dioxygenase activity at the MEJ. These aims will be elucidated using studies focused first on a floxed hemoglobin � mouse as well as a hemoglobin a over-expressing mouse to determine the effects of deletion/over-expression of this protein in endothelium on arterial reactivity, whole tissue blood flow, peripheral resistance and blood pressure. In addition, a human model of the disease alpha thalassemia where 2 alleles of hemoglobin a are deleted will be used to study the effects of this genome-wide heterozygous deletion on the vasculature. Next we will investigate how the hemoglobin � chaperone hemoglobin � stabilizing protein (AHSP) may traffic hemoglobin � to the myoendothelial junction or act directly as a regulator of the hemoglobin a redox state, altering the ability of nitric oxide to bind. The sum of this proposal unites and builds on the dat obtained from our previous R01 by allowing us for the first time to ask the direct question as to the function of the myoendothelial junction in intact animals. Indeed, we believe part of the answers could provide the basis for a completely new understanding of blood pressure control by the peripheral vasculature, as well as the derivation of unexplored pharmacological targets for control of hypertension.

描述（由适用提供）：我们最近发现血红蛋白富集在肌膜内皮连接处，这是内皮细胞和平滑肌细胞在抗药性动脉中接触的解剖位置。这是一个重要的发现，因为它表明血红蛋白在红细胞之外具有重要而积极的作用。该蛋白是仅有几种真正极化的蛋白质之一，该蛋白要定位于内皮衍生的肌膜内皮连接，siRNA诱导的蛋白质量的降低显着改变了动脉反应性，包括对苯肾上腺素和对乙酰胆碱的扩散量受到限制。我们得出的机制是基于证据，表明单体血红蛋白是一氧化氮的潜在清除剂，并且发现内皮一氧化氮合酶（ENOS）和血红蛋白在大分子配合物中。基于这项工作以及大量强的初步数据，我们假设血红蛋白在肌膜内皮结处是一种新的一氧化氮信号的调节剂，可能会影响血压调节。我们将使用两个具体目的检验这一假设：1。）研究内皮血红蛋白A基因消融/过表达对动脉功能的影响。2。）阐明AHSP和ENOS在MEJ上如何调节血红蛋白的表达和二氧酶活性。这些目标将使用首先集中于flox的血红蛋白小鼠以及过表达小鼠的血红蛋白的研究来阐明这些目标，以确定该蛋白在内皮中的缺失/过表达对动脉反应性，全组织血液流动，外周抵抗和血压的影响。此外，将使用2种血红蛋白A等位基因的疾病α丘脑贫血的模型，将使用该基因组杂合缺失的效果来研究血红蛋白的伴侣蛋白 - 稳定蛋白（AHSP）可能如何交通或直接动作肌素的蛋白质，或者是直接行动的，我们将如何处理血红蛋白的造成虫。血红蛋白A氧化还原态，改变了一氧化氮结合的能力。该提案单元的总和是基于从我们以前的R01获得的DAT，首次允许我们直接提出有关完整动物肌膜内皮交界的功能的直接问题。确实，我们认为部分答案可以为周围脉管系统对血压控制的全新理解以及对高血压控制的意外药物靶标的推导提供基础。

项目成果

Connexins in vascular physiology and pathology.

• DOI: 10.1089/ars.2008.2115
• 发表时间: 2009-02
• 期刊: Antioxidants & redox signaling
• 影响因子: 6.6
• 作者: Brisset AC;Isakson BE;Kwak BR
• 通讯作者: Kwak BR

AXL-Mediated Productive Infection of Human Endothelial Cells by Zika Virus.

• DOI: 10.1161/circresaha.116.309866
• 发表时间: 2016-11-11
• 期刊: Circulation research
• 影响因子: 20.1
• 作者: Liu S;DeLalio LJ;Isakson BE;Wang TT
• 通讯作者: Wang TT

Red Blood Cell Function and Dysfunction: Redox Regulation, Nitric Oxide Metabolism, Anemia.

• DOI: 10.1089/ars.2016.6954
• 发表时间: 2017-05-01
• 期刊: Antioxidants & redox signaling
• 影响因子: 6.6
• 作者: Kuhn V;Diederich L;Keller TCS 4th;Kramer CM;Lückstädt W;Panknin C;Suvorava T;Isakson BE;Kelm M;Cortese-Krott MM
• 通讯作者: Cortese-Krott MM

Loss of compliance in small arteries, but not in conduit arteries, after 6 weeks exposure to high fat diet.

暴露于高脂肪饮食 6 周后，小动脉顺应性丧失，但导管动脉没有顺应性丧失。

• DOI: 10.1007/s12265-012-9354-y
• 发表时间: 2012
• 期刊: Journal of cardiovascular translational research
• 影响因子: 3.4
• 作者: Billaud,Marie;Johnstone,ScottR;Isakson,BrantE
• 通讯作者: Isakson,BrantE

Expression of pannexin isoforms in the systemic murine arterial network.

• DOI: 10.1159/000338758
• 发表时间: 2012
• 期刊: Journal of vascular research
• 影响因子: 1.7
• 作者: Lohman AW;Billaud M;Straub AC;Johnstone SR;Best AK;Lee M;Barr K;Penuela S;Laird DW;Isakson BE
• 通讯作者: Isakson BE