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 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。我们的目标是使用模型系统研究基质金属蛋白酶 (MMP) 在血管生成和血管生成中的作用，其中真皮成纤维细胞与成血管细胞（CD34+干细胞 - 血管生成模型）或分化的血管内皮细胞（CD34+干细胞 - 血管生成模型）在 3 维基质中共培养（ HUVEC-血管生成模型）。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3维基质（胶原蛋白、基质胶、纤维蛋白）。此外，还将使用由胶原蛋白、硫酸软骨素和壳聚糖组成的复合基质，该基质已成功用于血管组织工程。该提议基于的观察结果表明，多种 MMP 有时可能在内皮细胞分化、血管形成和血管维持中发挥相反的作用。我们假设不同的 MMP 将参与血管形成，具体取决于实验中使用的是内皮细胞还是干细胞以及 3 维基质的选择。具体来说，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 3 维基质中形成毛细血管样管状结构。 2a) 使用通用 MMP 抑制剂来确定 MMP 是否参与这些结构的形成（和维持）。 2b) 识别 HUVEC 和 CD34+ 干细胞毛细血管发育过程中存在的特定 MMP。 2c) 使用慢病毒载体传递的稳定 mRNA 沉默来确定相同的 MMP 是否在功能上参与 HUVEC 和 CD34+ 干细胞的毛细血管形成。 3) 使用 DNA 微阵列分析来鉴定参与 HUVEC 和/或 CD34+ 干细胞毛细血管形成的新基因，并且当总 MMP 活性受到抑制或特定 MMP 的表达被阻断时，这些基因的表达会发生改变。我们的实验将揭示对血管生成/血管发生中涉及的冗余细胞类型和分子机制的更好理解，这将有助于理清不同细胞类型、不同MMP和不同ECM蛋白在血管形成中所起的作用。通过这种方式，我们的研究可能会带来临床应用，包括：阻断肿瘤血管生成、促进心肌梗塞的血管化以及形成组织工程血管移植物。