STRUCTURE-FUNCTION RELATIONSHIPS OF HEPARAN SULFATES IN VASCULOGENESIS

硫酸乙酰肝素在血管生成中的结构-功能关系

• 批准号: 8365540
• 负责人: MATTHEW A NUGENT
• 金额: $ 2万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365540
• 关键词: Affinity; Algorithms; Aliquot; Aorta; Atherosclerosis; Binding; Biological Assay; Biology; Blood Vessels; Cardiovascular Diseases; Cause of Death; Cells; Cellular Assay; Cessation of life; Collaborations; Data; Disaccharides; Disease; Endothelium; Engineering; Enzymes; Event; Extracellular Matrix; FGF2 gene; Fibroblast Growth Factor; Fibroblast Growth Factor 2; Fibroblast Growth Factor Receptors; Fluorescence; Functional disorder; Funding; Grant; Growth Factor; Heparin; Heparitin Sulfate; Inflammatory; Injury; Inorganic Sulfates; Lyase; Mass Spectrum Analysis; Mediating; Medicine; Methods; National Center for Research Resources; Oligosaccharides; Population; Principal Investigator; Process; Production; Rattus; Reader; Relative (related person); Research; Research Infrastructure; Resources; Response to stimulus physiology; Role; Series; Signal Transduction; Smooth Muscle Myocytes; Source; Stem cells; Stimulation of Cell Proliferation; Structure; Structure-Activity Relationship; Tissues; United States National Institutes of Health; Unspecified or Sulfate Ion Sulfates; cost; injured; interest; liquid chromatography mass spectrometry; receptor expression; repaired; response; restenosis; vasculogenesis

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Cardiovascular disease is the number one cause of death in the United Sates, and atherosclerosis and restenosis are responsible for the majority of these deaths. Neointimal expansion in atherosclerotic blood vessels and during restenosis is characterized by a series of events that reflect an attempt of the resident vascular cells to repair the dysfunctional blood vessel wall. Events such as: phenotypic changes in vascular smooth muscle cells leading to altered production of extracellular matrix, activation and recruitment of inflammatory cells, and recruitment of circulating endothelial progenitor cells reflect an attempt to effectively repair the blood vessel wall by establishing a functioning endothelium. However, in disease these processes may be insufficient or not properly controlled leading to a cascading propagation of the "injured/inflamed" state and continued vascular dysfunction. At the center of this stimulus-response network is the extracellular matrix (ECM), which both regulates and is regulated by the resident cells in the blood vessel wall. In this project we will define the central role of the ECM as a regulator of vascular growth factor activity. This collaboration involves functional characterization of heparan sulfate (HS) populations that bind fibroblast growth factors (FGFs). The activities of HS oligosaccharides of interest are being probed using a BaF32 cellular assay. The Baf32 cells are deficient in HS synthesis and lack FGF and FGF receptor expression. The cells have been engineered to express FGF receptor and will undergo FGF-mediated mitogenesis only when both FGF and heparin/HS are present. This assay has been scaled down for analysis using a fluorescence multiwall reader. Using this approach, it will be possible to determine the ability of defined HS fractions to induce or inhibit mitogenesis, as appropriate. Progress in 2010: The BaF32 assay was used to demonstrate that HS dp6 that bound to FGF2 showed increased activity in potentiating FGF2 signaling response relative to control dp6 (1). We are now using the glycomics methods developed during previous years to determine the changes to HS expression in aorta during balloon injury. HS was extracted from normal and injured rat aorta tissue. An aliquot of the HS was subjected to disaccharide analysis using a series of lyase enzymes to produce information on HS domain structure. The data will be analyzed using an algorithm "ChamP" developed by Nugent and co-workers at BUSM. In addition, we are acquiring LC/MS data on HS oligosaccharides corresponding to N-sulfated domains so as to extend the structural detail on HS structure during injury. 1. Naimy, H., Buczek-Thomas, J. A., Nugent, M. A., Leymarie, N., and Zaia, J. (2011) Highly sulfated non reducing end-derived heparan sulfate domains bind fibroblast growth factor-2 with high affinity and are enriched in biologically active fractions., J Biol Chem accepted 3/24/11.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 心血管疾病是联合病人的第一名死亡原因，动脉粥样硬化和再狭窄造成了大多数这些死亡。 动脉粥样硬化血管和再狭窄期间的新内膜扩张的特征是一系列事件，反映了居民血管细胞修复功能失调的血管壁的尝试。例如：血管平滑肌细胞的表型变化，导致细胞外基质的产生，炎症细胞的激活和募集的产生，以及募集循环内皮祖细胞的募集，反映了试图通过建立功能性内皮的有效修复血管壁的尝试。但是，在疾病中，这些过程可能不足或无法正确控制，从而导致“受伤/发炎”状态的层叠式传播和持续的血管功能障碍。 该刺激反应网络的中心是细胞外基质（ECM），既由血管壁中的居民细胞调节并调节。在这个项目中，我们将定义ECM作为血管生长因子活性的调节剂的核心作用。 这种合作涉及结合成纤维细胞生长因子（FGF）的乙par（HS）种群的功能表征。 使用BAF32细胞测定法对HS寡糖的活性进行了探测。 BAF32细胞缺乏HS合成，缺乏FGF和FGF受体表达。 这些细胞已经过设计以表达FGF受体，并且只有在存在FGF和肝素/HS时才会进行FGF介导的有丝分裂发生。 该测定法已缩小使用荧光多壁读取器进行分析。 使用这种方法，可以确定定义的HS级分诱导或抑制有丝分裂发生的能力。 2010年进度： BAF32测定法证明与FGF2结合的HS DP6在相对于对照DP6（1）的FGF2信号响应中显示出增加的活性。 现在，我们正在使用前几年开发的糖基因方法来确定气球损伤期间主动脉表达的变化。 从正常和受伤的大鼠主动脉组织中提取HS。 使用一系列裂解酶对HS结构结构的信息进行了二糖分析，对HS进行了二糖分析。 将使用Nugent和Busm的同事开发的算法“冠军”对数据进行分析。 此外，我们正在获取与N-硫化结构域相对应的HS寡糖的LC/MS数据，以扩展损伤期间HS结构的结构细节。 1。Naimy，H.，Buczek-Thomas，J.A.，Nugent，M.A.，Leymarie，N。和Zaia，J。（2011）高度硫化的非还原性的非还原终端源自硫酸盐硫酸盐结构剂结合了成纤维母细胞生长因子-2与高亲和力与高亲和力与生物学活跃的分裂富集，并在生物学上具有活性分裂。