Regulatory Roles for Vascular Peptidases in Angiogenesis

血管肽酶在血管生成中的调节作用

• 批准号: 7260512
• 负责人: RENATA PASQUALINI
• 金额: $ 35.56万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7260512
• 关键词: Accounting; Angiogenic Factor; Angiotensin III; Binding; Biochemical Markers; Biological Assay; Biology; Blood Vessels; Cell Line; Cell membrane; Cell surface; Cellular biology; Complex; Development; Disease; End Point; Endopeptidases; Endothelial Cells; Enzymes; Extracellular Matrix; Fingerprint; Genetic; Growth Factor; Hypoxia; In Situ; In Vitro; Inflammation; Knockout Mice; Lead; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Membrane; Methodology; Modeling; Molecular; Mus; Myocardial Infarction; Nature; Oxygen; Pathologic Neovascularization; Peptide Hydrolases; Peptide antibodies; Peptides; Pericytes; Phenotype; Physiological; Play; Process; Regulatory Pathway; Reporting; Retinal; Retinal Diseases; Role; Staging; Staining method; Stains; Stimulus; Stromal Cells; Tumor-Associated Vasculature; Up-Regulation; Vascularization; alanine aminopeptidase; angiogenesis; angiotensin II-producing serum enzyme III; base; cytokine; embryonic stem cell; enzyme substrate; homologous recombination; human tissue; hypoxia neonatorum; in vivo; inhibitor/antagonist; insight; mouse model; neoplastic cell; neovascularization; novel therapeutics; research study; response; therapy development; tumor

DESCRIPTION (provided by applicant): Angiogenesis is a complex multi-step process that can occur in development and diseases with a vascular component in response to various stimuli. Many lines of evidence indicate that angiogenesis depends on proteolytic activity. Activated blood vessels express biochemical markers that are differentially expressed and whose functional importance tias just begun to be uncovered. While only a few markers associated with angiogenic blood vessels have thus far been reported, it is remarkable that several are cell membrane-associated proteinases. By using genetic elimination and Diochemical inhibition, we have uncovered an as yet unrecognized mechanistic role for APA and CD13/APN in pathological angiogenesis. We hypothesize that APA and CD 13 activity contribute to important regulatory pathways. Here, we propose to investigate the mechanisms by which CD 13 and APA expression and activity control the physiological and pathological proliferation of activated endothelial cells forming blood vessels. First, we will study the induction and activity of CD 13 and APA during angiogenesis. We will evaluate the expression of CD 13 and of APA in angiogenic vasculature. As cell surface, membrane-bound enzymes upregulated in tumor vessels, APA and CD 13 may contribute to the initiation and/or progression of angiogenesis by (i) degrading inhibitory peptides, (ii) activating stimulatory molecules, or both. We will establish whether or not the levels of peptides known to bind and be degraded by APA and CD 13 are altered in activated blood vessels. Our main focus will be on the interdependent substrates for these enzymes, angiotensin III and VI. Angiotensin III has been shown to be pro-angiogenic, supporting our hypothesis. Second, we will determine the nature of the angiogenic stimuli leading to upregulation and activation of CD13/APN and APA in endothelial cells and pericytes. We will also examine the effects of inhibitors of CD13/APN and APA in angiogenesis. We will utilize anti-CD 13 and anti-APA inhibitory antibodies and peptides to evaluate if these peptidases enhance/accelerate angiogenesis ex-vivo and in vivo. Finally, we will evaluate the phenotype of CD13 and APA deficient mice with respect to angiogenesis using cytokine-, oxygen-and tumor-induced angiogenesis models. It is our hypothesis that these peptidases play complementary roles in angiogenesis. Our studies are likely to establish a mechanistic basis for the role of pro-angiogenic peptidases in the formation and maintenance of neovasculature, an aspect of high relevance in vascular biology. The proposed experiments in this application may also lead to development of new therapeutic strategies for diseases with an angiogenic component such as cancer and retinopathies.

描述（由申请人提供）：血管生成是一个复杂的多步骤过程，可能发生在发育和疾病中，其中血管成分响应各种刺激。许多证据表明血管生成取决于蛋白水解活性。活化的血管表达差异表达的生化标志物，其功能重要性刚刚开始被发现。虽然迄今为止只报道了少数与血管生成相关的标记物，但值得注意的是其中一些标记物是细胞膜相关的蛋白酶。通过使用遗传消除和生物化学抑制，我们发现了 APA 和 CD13/APN 在病理性血管生成中尚未认识到的机制作用。我们假设 APA 和 CD 13 活性有助于重要的调控途径。在这里，我们建议研究 CD 13 和 APA 表达和活性控制形成血管的活化内皮细胞的生理和病理增殖的机制。首先，我们将研究血管生成过程中 CD 13 和 APA 的诱导和活性。我们将评估血管生成脉管系统中 CD 13 和 APA 的表达。作为细胞表面，肿瘤血管中上调的膜结合酶、APA 和 CD 13 可能通过 (i) 降解抑制肽、(ii) 激活刺激分子或两者来促进血管生成的启动和/或进展。我们将确定已知与 APA 和 CD 13 结合并被降解的肽水平在活化的血管中是否发生改变。我们的主要关注点是这些酶的相互依赖的底物，血管紧张素 III 和 VI。血管紧张素 III 已被证明具有促血管生成作用，支持了我们的假设。其次，我们将确定导致内皮细胞和周细胞中 CD13/APN 和 APA 上调和激活的血管生成刺激的性质。我们还将检查 CD13/APN 和 APA 抑制剂在血管生成中的作用。我们将利用抗 CD 13 和抗 APA 抑制性抗体和肽来评估这些肽酶是否增强/加速离体和体内血管生成。最后，我们将使用细胞因子、氧气和肿瘤诱导的血管生成模型评估 CD13 和 APA 缺陷小鼠的血管生成表型。我们假设这些肽酶在血管生成中发挥互补作用。我们的研究可能为促血管生成肽酶在新血管系统的形成和维持中的作用建立机制基础，这是血管生物学中高度相关的一个方面。本申请中提出的实验还可能导致针对具有血管生成成分的疾病（例如癌症和视网膜病）开发新的治疗策略。