SIGNALING MECHANISMS IN REGULATION OF TUMOR ANGIOGENESIS

肿瘤血管生成调节的信号机制

基本信息

批准号：
8279120
负责人：
Dominic E. Cosgrove
金额：
$ 26.45万
依托单位：
FATHER FLANAGAN'S BOYS' HOME
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8279120
关键词：
Adenoviruses Affect Angiogenesis Inhibitors Angiogenic Factor Apoptosis Apoptotic Applications Grants Baculovirus Expression System Basement membrane Binding Binding Sites Biochemical Pathway Blood Blood Vessels C-terminal Catabolism Catalytic Domain Cause of Death Cell Culture System Cell Culture Techniques Cell Proliferation Cell surface Cells Cessation of life Co-Immunoprecipitations Collagen Type IV Colon Carcinoma Complex Data Development Disease Endothelial Cells Extracellular Matrix Fibroblast Growth Factor 2 Gelatinase A Goals Immunoblotting Immunohistochemistry Integrins Investigation Laboratories Life Lung MAP Kinase Gene MMP14 gene Malignant Neoplasms Mediating Mitochondria Modeling Molecular Molecular Target Mus PTGS2 gene PTK2 gene Pathway interactions Phase Phosphorylation Plasma Platelet-Derived Growth Factor Proliferating Proteins Regulation Role Signal Pathway Signal Transduction Solid Neoplasm Stream Testing Therapeutic Transgenic Mice Tumor Angiogenesis Tumor Necrosis Factor Ligand Superfamily Member 6 Up-Regulation Vascular Endothelial Growth Factors Work angiogenesis base bevacizumab cancer type caspase-3 in vivo inhibitor/antagonist insight matrigel migration mouse model proMMP-2 public health relevance therapeutic target tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Cancer is currently one of the most prevalent causes of death in the U.S.A. A major therapeutic option aims to slow the progression of cancer (example: use of anti-VEGF in colon cancer). Therefore, a renewed effort must be made to identify relevant molecular pathways, which could be exploited as therapeutic targets. Progression of many types of cancers is associated with up-regulation of several proangiogenic factors (HIF- 1a, VEGF, bFGF, PDGF, and MMP's etc.) which promotes tumor angiogenesis. These angiogenic factors activate different signaling pathways that regulate cell proliferation, migration and apoptosis. The long-term goals of my laboratory aims to understand the complex signaling mechanisms of angioinhibitors from extracellular matrix (type IV collagen NC1 domains, which are normally present in the blood), and to determine their role in antiangiogenic signaling. Recently my laboratory determined that a1(IV)NC1 mediates multiple signaling mechanisms to regulate tumor angiogenesis. However, its complex influences on tumor- angiogenesis and tumor growth are far from being fully understood. The present grant proposal is specifically directed at elucidating the mechanism whereby a1(IV)NC1 inhibits expression of the pro-angiogenic molecules, and its role in inhibition of tumor growth. In this study we aim to test our hypothesis: "a1(IV)NC1 promotes endothelial cell apoptosis through a1¿1 integrin and inhibits MMP-2 activation through an integrin independent pathway. These activities are critical for the antiangiogenic and antitumorogenic activity of a1(IV)NC1". Towards this end, we have generated an a1(IV)NC1 transgenic mouse model and adenoviruses, cloned and expressed a1(IV)NC1 in the baculovirus expression system and will use this protein for studies outlined in the specific aims. (1) Investigate the angioinhibitory signaling mechanisms by which a1(IV)NC1 inhibits the expression of angiogenic factors, including investigation of possible biochemical pathways. (2) Investigate the complex interactions between a1(IV)NC1/MMP-2 and identify the complementary binding sites involved in this interaction and function. (3) To investigate inhibition of tumor growth by a1(IV)NC1 in a1-integrin null and a1(IV)NC1 transgenic mice. The above Specific aims will be accomplished using cell cultures, a1 integrin null, a1(IV)NC1 transgenic mice and adenoviruses harboring a1(IV)NC1 domain expression cassettes. We will analyze these models using immunoblotting, co-immunoprecipitation, immunohistochemistry, Matrigel plug, and tumor studies. Successful completion of the proposed work will provide valuable insights that may facilitate the development of new antiangiogenic tumor therapies. PUBLIC HEALTH RELEVANCE: Cancer is currently one of the most prevalent causes of death in the USA, and current therapeutic options aim only to slow the progression of cancer. Therefore, a renewed effort must be made to identify nontoxic endogenous circulating anticancer molecules which could be exploited as therapeutic targets. My laboratory has cloned and expressed one such circulating molecule, and is presently testing it in a cell culture system and in live mice having tumors. The data being developed from these proposed studies have potential applications to cure solid tumor growth (cancers) in which angiogenesis contributes to the disease phase.

描述（由适用提供）：癌症目前是美国最普遍的死亡原因之一。一种主要的治疗选择旨在减缓癌症的进展（例如：在结肠癌中使用抗VEGF）。因此，必须采取新的努力来识别相关的分子途径，这可以被用作治疗靶标。多种类型的癌症的进展与促进肿瘤血管生成的几种促血管生成因子（HIF-1A，VEGF，BFGF，PDGF和MMP等）的上调有关。这些血管生成因子激活了调节细胞增殖，迁移和凋亡的不同信号通路。我的实验室的长期目标旨在了解细胞外基质（通常存在于血液中的IV型胶原蛋白NC1域）中血管生成的复杂信号传导机制，并确定其在抗高基因信号传导中的作用。最近，我的实验室确定A1（IV）NC1介导了多种信号传导机制以调节肿瘤血管生成。然而，它对肿瘤血管生成和肿瘤生长的复杂影响远非充分理解。目前的赠款提案专门阐明A1（IV）NC1抑制促血管生成分子的表达及其在抑制肿瘤生长中的作用的机制。在这项研究中，我们旨在检验我们的假设：“ A1（IV）NC1通过A1¿1整联蛋白促进内皮细胞凋亡，并通过整合素独立途径抑制MMP-2激活。这些活性对于A1（IV）NC1的抗抗生物和抗肿瘤活性至关重要。为此，我们已经在杆状病毒表达系统中生成了A1（IV）NC1转基因小鼠模型和腺病毒，并将A1（IV）NC1克隆并表达，并将使用该蛋白质用于特定目的中概述的研究。（1）研究A1（IV）NC1抑制血管生成因子的表达，包括研究可能的生化途径的表达，研究了血管抑制信号传导机制。（2）研究A1（IV）NC1/MMP-2之间的复杂相互作用，并确定与此相互作用和功能有关的互补结合位点。（3）研究A1-整合素无效的A1（IV）NC1对肿瘤生长的抑制作用和A1（IV）NC1转基因小鼠。上述特定目标将使用细胞培养物A1整合蛋白NULL，A1（IV）NC1转基因小鼠和带有A1（IV）NC1域表达盒的腺病毒。我们将使用免疫印迹，共免疫沉淀，免疫组织化学，母质塞和肿瘤研究来分析这些模型。成功完成拟议的工作将提供有价值的见解，以支持新的抗血管生成肿瘤疗法的发展。公共卫生相关性：癌症目前是美国最普遍的死亡原因之一，目前的治疗选择只旨在减缓癌症的进展。因此，必须采取新的努力来识别无毒的内源性循环抗药性分子，该分子可以作为治疗靶标探索。我的实验室已经克隆并表达了一个循环分子，目前正在细胞培养系统和具有肿瘤的活小鼠中进行测试。这些研究从这些拟议的研究中得出的数据具有潜在的应用来治疗血管生成有助于疾病阶段的实体瘤生长（癌症）。