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）：这项新申请解决了血管生物学中的一个重要问题，该问题涉及如何稳定新形成的内皮细胞（EC）衬里的管。我们在体外开发了这一过程的优秀模型，并确定了这些事件所需的关键机制和分子。 EC 衍生的蛋白酶（例如 MMP-1）能够在肾小管形态发生过程中降解 3D 胶原基质，从而破坏肾小管的稳定性并诱导退化。我们假设 EC 支持细胞（如周细胞）存在蛋白酶抑制剂（如 TIMP-3 和 RECK），其阻止 EC 在血管生成过程中降解其悬浮的细胞外基质环境的自然趋势。这些周细胞衍生的抑制剂可能代表对新形成的管的稳定至关重要的新分子。初步数据表明，TIMP-3 的独特之处在于它可以完全抑制 EC 肾小管形态发生并稳定 EC 内衬管。此外，TIMP-3 在周细胞的原代培养物中大量表达，而 EC 不表达。将血管平滑肌细胞（也表达 TIMP-3）与 EC 纳入 MMP-1 依赖性管回归的 3D 模型中，通过干扰 MMP-1 酶原激活来完全阻止该过程。我们将利用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）。周细胞在血管生成和血管发育过程中如何稳定 EC 管的机制仍然未知，该提议将直接检验蛋白酶抑制剂是调节这些事件的关键分子的新假设。 该应用程序的具体目标是； 目标#1。研究调节周细胞/血管平滑肌细胞在体外和体内诱导毛细管稳定的能力的主要机制和分子（即蛋白酶抑制剂和生长因子）。 目标#2。旨在研究 TIMP-3 在三维基质中的肾小管形态发生过程中通过完全抑制 EC 侵袭和管腔发育来调节体外和体内毛细管稳定性的分子机制。 目标#3。研究人基质金属蛋白酶-1 (MMP-1) 以及小鼠间质胶原酶（例如 MMP-13）在体外和体内直接控制毛细管退化事件的能力。