ROS Mechanisms in BAV Aortopathy

BAV 主动脉病中的 ROS 机制

• 批准号: 9058113
• 负责人: Thomas Gillette Gleason
• 金额: $ 52万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9058113
• 关键词: 3-Dimensional; Age; Aneurysm; Aortic Aneurysm; Architecture; Biochemistry; Biomechanics; Blood Vessels; Caliber; Caring; Cause of Death; Cell Survival; Cells; Clinical; Collagen; Collagen Fiber; Confocal Microscopy; Congenital Abnormality; Data; Detection; Development; Diagnostic; Dissection; Early Diagnosis; Ehlers-Danlos Syndrome; Elastin; Elastin Fiber; Electron Spin Resonance Spectroscopy; Enzymatic Biochemistry; Exhibits; Extracellular Matrix; Fluorescence; Generations; Genes; Goals; Heart Abnormalities; High Pressure Liquid Chromatography; Histology; Hospital Mortality; Human; Image; In Vitro; Individual; Knowledge; Matrix Metalloproteinases; Mediating; Microscopy; Modeling; Molecular; Morphology; Mutation; Operative Surgical Procedures; Oxidative Stress; Patients; Phenotype; Population; Property; Reactive Oxygen Species; Research Design; Research Personnel; Risk; Rupture; Smooth Muscle Myocytes; Specimen; Spin Trapping; Staging; Techniques; Testing; Thoracic Aortic Aneurysm; Time; Tissue Banking; Tissue Banks; United States; adjudicate; aortic valve; ascending aorta; base; bicuspid aortic valve; biological adaptation to stress; cohort; febuxostat; improved; inhibitor/antagonist; innovation; mortality; multi-photon; novel strategies; prevent; scaffold; second harmonic; sex; tissue culture; tool

DESCRIPTION (provided by applicant): This project will test the hypothesis that reactive oxygen species (ROS) accumulation in aortic smooth muscle cells (SMCs) mediates the aortopathy in patients with bicuspid aortic valve (BAV). The interdisciplinary investigative team possesses a unique composition of expertise and has devised a novel approach to prove that ROS accumulation in BAV-ascending aortic aneurysm (TAA) specimens leads to de-differentiation of SMCs, abnormal extracellular matrix (ECM) composition and architecture and altered biomechanical strength of the aortic wall. The hypothesis is supported by the team's strong preliminary data demonstrating ROS accumulation, reduced cell viability in the presence of ROS, diminished oxidative stress responses, de- differentiation of SMCs, disrupted matrix architecture and altered biomechanical tensile and delamination strengths in the ascending aorta of BAV patients compared with tricuspid aortic valve (TAV)-TAA and non- aneurysmal patients. The innovative strategy is accomplished in a two-aim approach: 1) Define what changes in SMC phenotype, ECM composition and architecture, and biomechanical tensile and delamination strengths are associated with ROS accumulation in BAV aortopathy and 2) Prove that ROS accumulation in aortic SMCs mediates the BAV aortopathy. This study will exploit 1) the PI's extensive tissue bank of human ascending aortic specimens and primary SMCs isolated from the following patient cohorts: a) BAV-TAA compared with b) non-aneurysmal BAV, c) TAV-TAA, and d) non-aneurysmal TAV "normal"; and 2) the team's established scaffold-based 3-D tissue culture model. ROS will be evaluated using the investigators' expertise in electron paramagnetic resonance spin trapping and confocal microscopy of fluorescence-based ROS probes. Innovative assessment of ECM composition and architecture will be achieved using multi-photon microscopy with second harmonic generation to define alignment of collagen and elastin fibers, histological detection of collagen and elastin, and quantification of MMP activity. The team's pioneering techniques for studying blood vessel biomechanics will be utilized throughout the research design. This study will, for the first time, define and prove that ROS mechanisms mediate the BAV aortopathy. The results will potentiate significant long-term clinical benefits including the development of improved diagnostic tools for earlier detection and better surveillance of the BAV aortopathy and the discovery of pharmacologic therapies directed at modulation of ROS in the aortic wall to prevent aneurysm formation in BAV patients.

描述（由申请人提供）：该项目将检验以下假设：主动脉平滑肌细胞中的活性氧（ROS）积累会介导双刺主动脉瓣（BAV）患者的主动脉瘤。跨学科的调查团队具有独特的专业知识组成，并设计了一种新颖的方法来证明ROS在BAV刺激性主动脉瘤（TAA）中的积累会导致SMC，异常细胞外基质（ECM）组成（ECM）组成和建筑和建筑和改变的生物力学强度的差异化。 The hypothesis is supported by the team's strong preliminary data demonstrating ROS accumulation, reduced cell viability in the presence of ROS, diminished oxidative stress responses, de- differentiation of SMCs, disrupted matrix architecture and altered biomechanical tensile and delamination strengths in the ascending aorta of BAV patients compared with tricuspid aortic valve (TAV)-TAA and non- aneurysmal 患者。创新的策略是通过两种AIM方法来实现的：1）定义SMC表型，ECM组成和结构的变化，生物力学拉伸和分层强度与BAV主动脉病的ROS积累相关，而2）证明ROS在主动脉SMC中的ROS在主动脉smc中介导BAV Atrortopathy。这项研究将利用1）PI的广泛的人类升主标本的组织库和从以下患者群体中分离出来的主要SMC：a）BAV-TAA与b）b）b）b）b）bav-taa相比，b）非干扰性bav，c）tav-taa和d）d）d）nontheurysmal tav'; 2）团队建立的基于脚手架的3D组织培养模型。 ROS将使用研究者在电子顺磁共振自旋诱捕和基于荧光的ROS探针的共聚焦显微镜方面的专业知识进行评估。使用第二次谐波生成的多光子显微镜来实现ECM组成和结构的创新评估，以定义胶原蛋白和弹性蛋白纤维的比对，胶原蛋白和弹性蛋白的组织学检测以及MMP活性的定量。 在整个研究设计中，将利用该团队研究血管生物力学的开创性技术。这项研究将首次定义并证明ROS机制介导BAV主动脉病。该结果将增强长期的长期临床益处，包括开发改进的诊断工具，以便早期检测和更好地监测BAV主动脉病的监测以及发现针对主动脉壁调节ROS的药物治疗的发现，以防止BAV患者的动脉瘤形成。