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（由申请人提供）：癌症目前是美国最常见的死亡原因之一。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）。必须确定相关的分子途径，这些途径可以作为治疗靶标，许多类型的癌症的进展与几种促血管生成因子（HIF-1a、VEGF、bFGF、 PDGF和MMP等）促进肿瘤血管生成，这些血管生成因子激活调节细胞增殖、迁移和凋亡的不同信号通路。我实验室的长期目标是了解细胞外基质（类型）的血管抑制剂的复杂信号传导机制。 IV 胶原蛋白 NC1 结构域（通常存在于血液中），并确定其在抗血管生成信号传导中的作用。最近我的实验室确定 a1(IV)NC1 介导多种信号传导机制来调节肿瘤血管生成。对肿瘤血管生成和肿瘤生长的影响还远未完全了解，目前的拨款提案专门针对阐明 a1(IV)NC1 抑制促血管生成分子表达的机制及其在抑制肿瘤生长中的作用。在这项研究中，我们旨在检验我们的假设：“a1(IV)NC1 通过 a1¿1 整合素促进内皮细胞凋亡，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的抗血管生成和抗肿瘤活性”。为此，我们构建了a1(IV)NC1转基因小鼠模型和腺病毒，在杆状病毒表达系统中克隆并表达了a1(IV)NC1，并将将该蛋白用于具体目标中概述的研究 (1) 研究 a1(IV)NC1 抑制血管生成因子表达的血管抑制信号机制，包括可能的生化研究。 (2) 研究a1(IV)NC1/MMP-2 之间的复杂相互作用，并鉴定参与这种相互作用和功能的互补结合位点。 (3) 研究a1-整联蛋白无效时a1(IV)NC1 对肿瘤生长的抑制作用。上述具体目标将使用细胞培养物、a1整联蛋白缺失、a1(IV)NC1转基因小鼠和含有a1(IV)NC1结构域的腺病毒来实现。我们将使用免疫印迹、免疫共沉淀、免疫组织化学、基质胶塞和肿瘤研究来分析这些模型，成功完成拟议的工作将为新的抗血管生成肿瘤疗法的开发提供有价值的见解。公共健康相关性：癌症目前是美国最普遍的死亡原因之一，目前的治疗方案仅旨在减缓癌症的进展，因此，必须重新努力识别无毒的内源性循环抗癌分子。我的实验室已经克隆并表达了一种这样的循环分子，目前正在细胞培养系统和患有肿瘤的活小鼠中对其进行测试，从这些拟议研究中获得的数据具有治疗实体瘤生长的潜在应用。（癌症）其中血管生成导致疾病阶段。