MATRIX METALLOPROTEINASE IN VASCULOGENESIS & ANGIOGENESIS

血管发生中的基质金属蛋白酶

• 批准号: 7381238
• 负责人: PAL GOOZ
• 金额: $ 4.14万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7381238
• 关键词: angiogenesis; capillary; cell type; collagen; fibroblasts; model; play; role; stem cells; tissue engineering

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our objective is to study the role of matrix metalloproteinases (MMPs) in vasculogenesis and angiogenesis using a model system in which dermal fibroblasts are cocultured in 3-dimensional matrices along with hemangioblasts (CD34+ stem cells - vasculogenesis model) or with differentiated vascular endothelial cells (HUVEC -angiogenesis model). Different 3-dimensional matrices (collagen, Matrigel, fibrin) will be used that are similar to the different extracellular matrices that endothelial cells encounter in vivo. In addition, a composite matrix composed of collagen, chondroitin sulfate, and chitosan will be used that has already been successfully used in the tissue engineering of blood vessels. This proposal is based on observations suggesting that a variety of MMPs may play sometimes opposing roles in endothelial cell differentiation, the formation of vessels, and the maintenance of vessels. We hypothesize that different MMPs will be involved in vessel formation depending on whether endothelial cells or stems cells are used in experiments and depending on the choice of 3-dimensional matrix. Specifically we will: 1) Demonstrate that when CD 34+ stem cells enriched from peripheral blood mononuclear cells or HUVEC are co-cultured with dermal fibroblasts, capillary-like tubular structures form in 3-dimensional matrices. 2a) Use general MMP inhibitors to determine whether MMPs are involved in the formation (and maintenance) of these structures. 2b) Identify the specific MMPs that are present during the development of capillaries from HUVEC and CD34+ stem cells. 2c) Use stable mRNA silencing delivered by lentiviral vectors to determine whether the same MMPs are functionally involved in capillary formation by HUVEC and by CD34+ stem cells. 3) Use DNA microarray analysis to identify novel genes that are involved in capillary formation by HUVEC and/or CD34+ stem cells and whose expression is altered when total MMP activity is inhibited or when the expression of specific MMPs is blocked. The better understanding of the redundant cell types and molecular mechanisms involved in angiogenesis/vasculogenesis that will be revealed by our experiments will help sort out the roles that different cell types, different MMPs, and different ECM proteins play in vessel formation. In this manner, our studies may lead to clinical applications including: blocking tumor angiogenesis, promoting vascularization of myocardial infarcts, and forming tissue engineered vascular grafts.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们的目的是研究基质金属蛋白酶（MMP）在血管发生和血管生成中的作用，使用模型系统，其中真皮成纤维细胞与成血管细胞（CD 34+干细胞-血管发生模型）或与分化的血管内皮细胞（HUVEC -血管生成模型）共培养在三维基质中沿着。将使用与内皮细胞在体内遇到的不同细胞外基质相似的不同三维基质（胶原、基质胶、纤维蛋白）。此外，将使用由胶原蛋白、硫酸软骨素和壳聚糖组成的复合基质，该复合基质已经成功地用于血管的组织工程。这一建议是基于观察表明，各种基质金属蛋白酶有时可能发挥相反的作用，在内皮细胞分化，血管的形成，和维护的血管。我们假设不同的基质金属蛋白酶将参与血管形成，这取决于实验中是否使用内皮细胞或干细胞，并取决于三维基质的选择。具体而言，我们将：1）证明当从外周血单核细胞或HUVEC富集的CD 34+干细胞与真皮成纤维细胞共培养时，在三维基质中形成毛细血管样管状结构。2a）使用一般MMP抑制剂来确定MMP是否参与这些结构的形成（和维持）。2b）鉴定在来自HUVEC和CD 34+干细胞的毛细血管发育期间存在的特异性MMP。2c）使用由慢病毒载体递送的稳定mRNA沉默来确定相同的MMP是否在功能上参与HUVEC和CD 34+干细胞的毛细血管形成。3)使用DNA微阵列分析，以确定新的基因，参与毛细血管形成的HUVEC和/或CD 34+干细胞，其表达改变时，总MMP活性被抑制或特定MMP的表达被阻断。更好地了解参与血管生成/血管发生的冗余细胞类型和分子机制，将有助于我们的实验揭示不同细胞类型，不同MMP和不同ECM蛋白在血管形成中的作用。因此，我们的研究可能会导致临床应用，包括：阻断肿瘤血管生成，促进心肌梗死的血管化，形成组织工程血管移植物。