Pericyte Proteinase Inhibitors and EC Tube Stabilization

周细胞蛋白酶抑制剂和 EC 管稳定

• 批准号: 7163001
• 负责人: George E Davis
• 金额: $ 28.35万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7163001
• 关键词: Address; Angiopoietin-1; Appendix; Basement membrane; Biology; Blood Vessels; Blood capillaries; Cell Differentiation process; Cell Line; Cell Proliferation; Cell surface; Collagen; Data; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease regression; Disruption; Embryo; Endopeptidases; Endothelial Cells; Environment; Enzyme Precursors; Equilibrium; Event; Extracellular Matrix; Fibrin; Gelatinase A; Gelatinase B; Gene Delivery; Generations; Genes; Growth; Growth Factor; Heparitin Sulfate; Human; In Vitro; Interstitial Collagenase; Investigation; Kininogenase; Knockout Mice; Laminin; Left; Malignant Neoplasms; Matrix Metalloproteinases; Mechanics; Membrane; Metalloproteases; Modeling; Molecular; Morphogenesis; Mus; Pathogenesis; Pathologic Neovascularization; Peptide Hydrolases; Pericytes; Peripheral Vascular Diseases; Plasmin; Platelet-Derived Growth Factor; Play; Principal Investigator; Process; Production; Protease Inhibitor; Proteolysis; Reagent; Recombinants; Regulation; Reporting; Role; Serine Protease; Small Interfering RNA; Smooth Muscle Myocytes; Supporting Cell; Testing; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-3; Transforming Growth Factor beta; Trypsin; Tube; Tubular formation; Vascular Endothelial Growth Factors; Vascular Permeabilities; Vascularization; Withdrawal; angiogenesis; base; capillary; collagenase 3; concept; cytokine; human disease; in vitro Model; in vivo; inhibitor/antagonist; knockout gene; mechanical drive; neutralizing antibody; novel; response; scaffold; three-dimensional modeling; tissue-factor-pathway inhibitor 2; vasculogenesis

DESCRIPTION (provided by applicant): This new application addresses an important question in vascular biology which concerns how newly formed endothelial cell (EC)-lined tubes are stabilized. We have developed excellent models of this process in vitro and have identified key mechanisms and molecules, which are required for these events. EC-derived proteinases such as MMP-1 are capable of degrading 3D collagen matrices during tubular morphogenesis, which destabilizes tubes and induces regression. We hypothesize that EC supporting cells such as pericytes present proteinase inhibitors such as TIMP-3 and RECK which blocks the natural tendency of ECs during angiogenesis to degrade the extracellular matrix environment in which they are suspended. These pericyte-derived inhibitors likely represent new molecules that are critical to the stabilization of newly formed tubes. Preliminary data shows that TIMP-3 is unique in that it can completely inhibit EC tubular morphogenesis as well as stabilize EC-lined tubes. Furthermore, TIMP-3 is heavily expressed by primary cultures of pericytes and is not expressed by ECs. Inclusion of vascular smooth muscle cells (also express TIMP-3) with ECs in 3D models of MMP-1-dependent tube regression completely blocks the process by interfering with MMP-1 proenzyme activation. We will utilize a balanced experimental approach to determine the role of pericyte-derived proteinase inhibitors in EC tube stabilization in vitro and in vivo (using recombinant adenoviral gene delivery and TIMP-3 knockout mice). The mechanisms involved in how pericytes stabilize EC tubes during angiogenesis and vascular development remain unknown and this proposal will directly test the novel hypothesis that proteinase inhibitors are key molecules regulating these events. The specific aims of this application are; Aim #1. To investigate the primary mechanisms and molecules (i.e. proteinase inhibitors and growth factors) which regulate the ability of pericytes/vascular smooth muscle cells to induce capillary tube stabilization in vitro and in vivo. Aim #2. To investigate the molecular mechanism by which TIMP-3 regulates capillary tube stabilization in vitro and in vivo through complete inhibition of EC invasion and lumen development during tubular morphogenesis in three-dimensional matrices. Aim #3. To investigate the ability of human matrix metalloproteinase-1 (MMP-1) as well as mouse interstitial collagenases (e.g. MMP-13) to directly control capillary tube regression events in vitro and in vivo.

描述（由申请人提供）：此新应用程序解决了血管生物学中的一个重要问题，该问题涉及新形成的内皮细胞（EC）衬里的管子如何稳定。我们已经在体外开发了这一过程的出色模型，并确定了这些事件所需的关键机制和分子。在管状形态发生过程中，EC衍生的蛋白酶（例如MMP-1）能够降解3D胶原蛋白基质，从而破坏管子并诱导回归。我们假设EC支持细胞（如周细胞）呈现蛋白酶抑制剂，例如TIMP-3和RECK，它阻止了血管生成期间EC的自然趋势，从而降解了被悬浮的细胞外基质环境。这些衍生的周细胞抑制剂可能代表了对新形成的试管稳定至关重要的新分子。初步数据表明，TIMP-3是独一无二的，因为它可以完全抑制ec肾小管形态发生以及稳定EC衬管。此外，TIMP-3由周细胞的主要培养物大量表达，而不会由EC表达。在MMP-1依赖性管的3D模型中纳入血管平滑肌细胞（也表达EC），通过干扰MMP-1 Proeenzyme激活，完全阻止了该过程。我们将利用一种平衡的实验方法来确定周细胞衍生的蛋白酶抑制剂在体外和体内EC管稳定中的作用（使用重组腺病毒基因递送和TIMP-3敲除小鼠）。在血管生成和血管发育过程中，周细胞如何稳定EC管的机制尚不清楚，该建议将直接检验新的假设，即蛋白酶抑制剂是调节这些事件的关键分子。 该应用程序的具体目的是； 目标＃1。研究主要机制和分子（即蛋白酶抑制剂和生长因子），这些机制和分子抑制剂和生长因子）调节周细胞/血管平滑肌细胞在体外和体内诱导毛细管稳定的能力。 目标＃2。为了研究TIMP-3通过在三维矩阵中肾小管形态发生过程中的EC侵袭和流体发育中完全抑制EC的侵袭和流体发育，通过TIMP-3调节体外和体内毛细管稳定的分子机制。 目标＃3。研究人基质金属蛋白酶-1（MMP-1）以及小鼠间隙胶原酶（例如MMP-13）直接在体外和体内直接控制毛细管回归事件的能力。