Atheroprotective vs. Atherogenic Glycocalyx Mechanotransduction Mechanisms

动脉粥样硬化保护与致动脉粥样硬化糖萼机械转导机制

• 批准号: 9137702
• 负责人: Eno Essien Ebong
• 金额: $ 16.02万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-04 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9137702
• 关键词: Acute; Adhesions; Anatomy; Aneurysm; Animals; Apolipoprotein E; Atherosclerosis; Award; Binding; Biochemical; Biological Markers; Biomedical Engineering; Blood Vessels; California; Cardiovascular system; Carotid Arteries; Caveolae; Cell Culture Techniques; Cell Shape; Cell physiology; Cellular biology; Cities; Communication; Connexins; Core Protein; Cytoskeleton; Deacetylase; Development; Diagnosis; Disease; Electron Microscopy; Endothelial Cells; Engineering; Environment; Enzymes; Europe; Event; Faculty; Fatty acid glycerol esters; Freeze Substitution; Freezing; Functional disorder; Future; Gap Junctions; Geometry; Glycobiology; Glycocalyx; Glypican; Goals; Health; Heparitin Sulfate; Histologic; Immunofluorescence Microscopy; In Vitro; Institutes; Institution; Interdisciplinary Study; Knockout Mice; Laboratories; Lead; Lesion; Measures; Mediating; Medical; Medicine; Membrane; Mentored Research Scientist Development Award; Mentors; Microscopy; Modeling; Molecular Biology; Molecular Target; Morphology; Mus; Myocardial Infarction; New York; Nitric Oxide; Nitric Oxide Synthase; Operative Surgical Procedures; Pathology; Pattern; Peripheral Vascular Diseases; Positioning Attribute; Prevention approach; Production; Qualifying; Regulation; Relaxation; Research; Research Infrastructure; Research Personnel; Role; Severities; Signal Transduction; Site; Solid; Stimulus; Stroke; Structure; Surface; Techniques; Technology; Testing; Thick; Tissues; Training; Translating; Transmission Electron Microscopy; Uncertainty; United States; Universities; Vascular Diseases; Voice; Work; abstracting; atherogenesis; atheroprotective; career; cell behavior; cell type; cellular targeting; college; combat; constriction; experience; extracellular; feeding; improved; in vivo; in vivo Model; innovation; medical schools; meetings; method development; monolayer; mouse model; new therapeutic target; novel diagnostics; prevent; professor; programs; research facility; research study; response; structural biology; sulfotransferase; syndecan; tenure track; theories; tool; transmission process; vascular endothelial dysfunction

DESCRIPTION (provided by applicant): The K01 will facilitate Dr. Ebong's development of her independent research position as a tenure-track professor. Her research objective is to combine engineering with advanced microscopy, cell and molecular biology, and animal pathophysiology to elucidate endothelial cell (EC) glycocalyx (GCX) ultrastructure and its functional role in EC mechanotransduction and vascular function in health and atherosclerosis. With her background in core engineering theory and practice, and her solid interdisciplinary research experience that blends cardiovascular and cellular bioengineering with molecular biology, Dr. Ebong is highly qualified to carry out the Research Strategy and meet the Career Goals and Objectives. This award will enable her to expand her research, enhance its interpretation, and produce new discoveries. Dr. Ebong will be mentored by an interdisciplinary team that has broad expertise in bioengineering, in vitro and in vivo shear and mechanotransduction experiments, in vivo modeling of vascular pathology, glycobiology, microscopy, anatomy and structural biology, and molecular biology. The make-up of this team will provide her with an extensive network of techniques, facilities, and research institutions. The mentored activities proposed for this K01 program will be primarily conducted at Northeastern University and performed in Dr. Ebong's laboratory. Faculty and facilities of Northeastern (Drs. Ruberti, Khaw, and Hancock), The City College of New York (Dr. Tarbell), Albert Einstein College of Medicine (Mr. Macaluso), Emory University School of Medicine and Georgia Institute of Technology (Dr. Jo), Temple University School of Medicine (Dr. Rizzo), and University of California at San Diego (Dr. Esko) will also support Dr. Ebong's training. Atherosclerosis occurs at blood vessel sites, such as branches, where unstable (disturbed) flow makes the endothelial cell (EC) layer dysfunctional. Healthy tissue lines straight blood vessels where streamlined (uniform) flow permits normal EC function. The mechanisms by which flow alters EC function to prevent or promote atherosclerosis remain unclear. Defining these mechanisms is the primary focus of Dr. Ebong's research program. The K01 research strategy proposed by Dr. Ebong and her team is to study the structure and function of a prime EC mechanotransducer candidate-the surface glycocalyx (GCX)-which directly senses flow, can transmit force via the cytoskeleton to various biologically active cellulr sites, and sheds in atherosclerosis. GCX is essential for flow conversion to EC functions including nitric oxide (NO) release, cytoskeleton organization, gap junction communication, and cell shape that are abnormal in atherosclerosis. Dr. Ebong has already shown that flow induces vasoregulatory EC-type NO synthase (eNOS) activation, but only in the presence of the heparan sulfate (HS) glypican-1 (GPC-1) component of the EC GCX. Her work has revealed that flow-induced EC remodeling relies on the HS syndecan-1 (SDC-1) component of the GCX. Using rapid freezing/freeze substitution (RF/FS) transmission electron microscopy (TEM), she is currently defining the EC GCX ultrastructure and its changes due to cellular origin and the biochemical and flow environment. During the period of the K01 award, Dr. Ebong and her team will advance this work by testing the hypothesis that EC respond differently to uniform and disturbed flow through dynamic changes in GCX ultrastructure (Aim 1), which trigger the caveolae and cytoskeleton (and subsequently, the gap junctions and basal matrix) to differentially activate EC signaling and remodeling events (Aim 2) that lead to vascular health or disease (Aim 3). This hypothesis will be tested using cultured EC with intact or manipulated GCX, which are subjected to atheroprotective and atherogenic flow conditions that are replicated in vitro using a parallel plate flow chamber. Mouse models of GCX component deletion, acute disturbed flow, endothelial and vascular dysfunction, and atherosclerosis will also be employed. Flow-induced GCX thickness, morphology, and subcomponent content and organization will be assessed. Flow- and GCX-regulation of EC signaling and remodeling events such as eNOS activation and NO production, connexin-specific gap junction signaling, and remodeling of cell shape and basal adhesions will be examined. GCX component-specific involvement in EC- dependent vasoregulation, the development of robust atherosclerosis, and the determination of lesion complexity will also be studied. The team expects to identify GCX components that are biomarkers of atherosclerosis. The findings of this research will be essential for engineering new diagnostics and therapeutics that target the EC GCX to combat atherosclerosis. Dr. Ebong is talented, the mentoring and research team is experienced and interdisciplinary, and the research strategy is exciting and innovative. Without a doubt, Dr. Ebong will emerge from the K01 as an independent investigator with a unique identity, a wide-range of powerful research tools, a robust research infrastructure, and a voice in high-level discourse on GCX-related approaches to the prevention, diagnosis, and treatment of atherosclerosis. (End of Abstract)

描述（由申请人提供）：K01 将促进 Ebong 博士作为终身教授的独立研究职位的发展。她的研究目标是将工程学与先进的显微镜、细胞和分子生物学以及动物病理生理学相结合，阐明内皮细胞 (EC) 糖萼 (GCX) 超微结构及其在健康和动脉粥样硬化中 EC 机械转导和血管功能中的功能作用。凭借核心工程理论和实践的背景，以及将心血管和细胞生物工程与分子生物学相结合的扎实的跨学科研究经验，Ebong 博士非常有资格执行研究战略并实现职业目标和目标。该奖项将使她能够扩大研究范围，增强其解释并产生新的发现。 Ebong 博士将得到一个跨学科团队的指导，该团队在生物工程、体外和体内剪切和力转导实验、血管病理学体内建模、糖生物学、显微镜、解剖学和结构生物学以及分子生物学方面拥有广泛的专业知识。该团队的组成将为她提供广泛的技术、设施和研究机构网络。为该 K01 项目提议的指导活动将主要在东北大学进行，并在 Ebong 博士的实验室进行。东北大学（Ruberti、Khaw 和 Hancock 博士）、纽约城市学院（Tarbell 博士）、阿尔伯特·爱因斯坦医学院（Macaluso 先生）、埃默里大学医学院和佐治亚理工学院（Jo 博士）、天普大学医学院（Rizzo 博士）和加州大学圣地亚哥分校（Esko 博士）的教师和设施也将支持 Ebong 博士的培训。 动脉粥样硬化发生在血管部位，例如分支，不稳定（扰乱）的血流导致内皮细胞（EC）层功能障碍。健康的组织排列成笔直的血管，其中流线型（均匀）的血流允许正常的 EC 功能。血流改变 EC 功能以预防或促进动脉粥样硬化的机制仍不清楚。定义这些机制是 Ebong 博士研究项目的主要重点。 Ebong 博士和她的团队提出的 K01 研究策略是研究主要 EC 机械传感器候选物 - 表面糖萼 (GCX) 的结构和功能，它直接感知流动，可以通过细胞骨架将力传递到各种生物活性细胞位点，并在动脉粥样硬化中脱落。 GCX 对于血流转换为 EC 功能至关重要，包括一氧化氮 (NO) 释放、细胞骨架组织、间隙连接通讯和动脉粥样硬化中异常的细胞形状。 Ebong 博士已经证明，流动会诱导血管调节 EC 型 NO 合酶 (eNOS) 激活，但仅限于 EC GCX 的硫酸乙酰肝素 (HS) 磷脂酰肌醇蛋白聚糖-1 (GPC-1) 成分存在的情况下。她的工作表明，流动诱导的 EC 重塑依赖于 GCX 的 HS syndecan-1 (SDC-1) 成分。她目前正在使用快速冷冻/冷冻替代 (RF/FS) 透射电子显微镜 (TEM) 来定义 EC GCX 超微结构及其因细胞起源、生化和流动环境而产生的变化。 在获得 K01 奖期间，Ebong 博士和她的团队将通过测试以下假设来推进这项工作：EC 通过 GCX 超微结构的动态变化（目标 1）对均匀和扰动的流动做出不同的反应，从而触发小窝和细胞骨架（以及随后的间隙连接和基底基质）以差异性激活 EC 信号传导和重塑事件（目标 2），从而导致血管健康或疾病 （目标 3）。该假设将使用具有完整或操纵的 GCX 的培养 EC 进行测试，这些 EC 受到动脉粥样硬化和致动脉粥样硬化流动条件的影响，并使用平行板流动室在体外复制。还将采用 GCX 成分缺失、急性血流紊乱、内皮和血管功能障碍以及动脉粥样硬化的小鼠模型。将评估流动引起的 GCX 厚度、形态以及子成分含量和组织。将检查 EC 信号传导和重塑事件的流动和 GCX 调节，例如 eNOS 激活和 NO 产生、连接蛋白特异性间隙连接信号传导以及细胞形状和基底粘附的重塑。还将研究 GCX 成分特异性参与 EC 依赖性血管调节、强动脉粥样硬化的发展以及病变复杂性的确定。该团队希望鉴定出作为动脉粥样硬化生物标志物的 GCX 成分。这项研究的结果对于设计针对 EC GCX 对抗动脉粥样硬化的新诊断和治疗方法至关重要。 Ebong博士才华横溢，指导和研究团队经验丰富且跨学科，研究策略令人兴奋且创新。毫无疑问，Ebong 博士将从 K01 中脱颖而出，成为一名独立研究者，拥有独特的身份、广泛的强大研究工具、强大的研究基础设施，以及在与 GCX 相关的动脉粥样硬化预防、诊断和治疗方法的高层讨论中的发言权。 （摘要完）