Role of Inflammatory Neutrophils and their Unique MMP-9 in Tumor Progression

炎症性中性粒细胞及其独特的 MMP-9 在肿瘤进展中的作用

• 批准号: 8854046
• 负责人: JAMES P QUIGLEY
• 金额: $ 39.32万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8854046
• 关键词: Acute; Angiogenesis Inducing Agents; Angiogenic Factor; Angiogenic Switch; Biochemical; Blood Vessels; Bone Marrow; Bone Marrow Cells; Cells; Cessation of life; Chick Embryo; Chickens; Chronic; Endothelial Cells; Enzyme Precursors; Enzymes; Event; Gelatinase B; Goals; Human; Infection; Inflammation; Inflammatory; Injury; Kinetics; Laboratories; Light; Link; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Mesenchymal; Modeling; Monitor; Mus; Myeloid Cells; Nature; Neoplasm Metastasis; Neutrophil Infiltration; Nutrient; Organ; Pathologic Neovascularization; Physiologic Neovascularization; Plasmin; Plasminogen; Population; Primary Neoplasm; Process; Production; Prostate Adenocarcinoma; Prostate carcinoma; Proteomics; Reagent; Research; Role; Route; Serine Protease; Site; Squamous cell carcinoma; Staging; Stromal Cells; Stromal Neoplasm; Testing; Time; Tissues; Transgenic Model; Transgenic Organisms; Transplantation; Tumor Angiogenesis; Tumor Tissue; Variant; angiogenesis; cell type; comparative; congenic; cytokine; feeding; fibrosarcoma; granulocyte; human disease; in vivo; in vivo Model; in vivo imaging; knock-down; macrophage; monocyte; neoplastic cell; neovascular; neutrophil; research study; response; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): One of the early steps in malignant progression is tumor angiogenesis, supplying nutrients to the developing tumor and possibly providing routes for tumor cell escape. In numerous transplant and transgenic models, subsets of BMDCs were shown to deliver MMP-9 to mediate the angiogenic switch and frequently, the MMP-9-delivering BMDCs were identified as monocytes/macrophages. A few studies indicated that neutrophils might serve as an alternative BMDC type since they also express MMP-9 and have been associated, although transiently, with developing tumors. However, inflammatory neutrophils, with their only clear role in acute infection and injury, have generally been overlooked as contributors to tumor progression. We have recently demonstrated that neutrophils produce a unique, TIMP-free form of proMMP-9, making it a potent angiogenic agent. In light of this evidence and that neutrophils infiltrate tumor-stromal tissue and rapidly release their prestored, highly-angiogenic TIMP-free MMP-9, we would challenge the existing paradigms that monocyte-like cells are exclusive inducers of the angiogenic switch. We propose as an overall hypothesis that the initial delivery of neutrophils and their unique MMP-9 is a critical and targetable early event in tumor progression. To test this hypothesis we propose the following Specific Aims: Aim 1: To Directly Compare the Angiogenic Capabilities of Inflammatory Neutrophils and Their Unique TIMP-Free MMP-9 with Differentiating Monocytes and Their Secreted MMP-9. Comparisons will include kinetic studies of MMP-9 release/production by the different myeloid cell types, influx of MMP-9-delivering inflammatory cells into angiogenic and tumor tissues, and quantifying the in vivo angiogenic activity of MMP-9 delivered by inflammatory neutrophils versus monocytes. Aim 2: To Elucidate the Biochemical Mechanism(s) which Determine the High Pro-Angiogenic and Pro-Tumor Progression Potentials of Neutrophil MMP-9. The inducers of neutrophil influx into tumor tissues will be quantified. The mechanism of activation of the TIMP-free neutrophil MMP-9 zymogen will be determined, which in vivo has never been documented. The plasminogen/plasmin serine protease cascade will be examined mechanistically for partnering with activated MMP-9 during angiogenic induction. The in vivo substrates and effector molecules of activated neutrophil MMP-9 will be probed by new proteomic approaches. Aim 3: To Validate That the Pro-Angiogenic Activity of Neutrophil MMP-9 is Causally Linked to Tumor Dissemination via Neovascular Conduits. The specific reduction or stimulation of neutrophil MMP-9-mediated angiogenesis will be related directly to the levels of tumor cell intravasation and confirmed with in vivo rescue experiments using viable neutrophils and/or purified neutrophil MMP-9. The results will be then validated in a number of murine models of angiogenesis and metastasis including orthotopic transplants and transgenic models focusing on the human disease-like TRAMP model.

描述(申请人提供)：恶性进展的早期步骤之一是肿瘤血管生成，为发展中的肿瘤提供营养物质，并可能为肿瘤细胞逃逸提供途径。在众多的移植和转基因模型中，BMDCs的亚群被证明能够传递基质金属蛋白酶-9来介导血管生成的转换，并且通常，传递基质金属蛋白酶-9的骨髓基质细胞被鉴定为单核/巨噬细胞。一些研究表明，中性粒细胞可能是一种替代的BMDC类型，因为它们也表达基质金属蛋白酶-9，并与肿瘤的发生有关，尽管是暂时的。然而，炎性中性粒细胞在急性感染和损伤中唯一明确的作用，通常被忽视为肿瘤进展的贡献者。我们最近证明，中性粒细胞产生一种独特的、不含TIMP的原基质金属蛋白酶-9，使其成为一种有效的血管生成剂。鉴于这一证据以及中性粒细胞渗入肿瘤间质组织并迅速释放其预先储存的、高度血管生成的无TIMP的基质金属蛋白酶-9，我们将挑战现有的单核细胞样细胞是血管生成开关的唯一诱导者的范式。我们提出了一个总体假设，即中性粒细胞及其独特的基质金属蛋白酶-9的最初交付是肿瘤进展中一个关键的和有针对性的早期事件。为了验证这一假说，我们提出了以下具体目标：目的1：直接比较炎性中性粒细胞及其特有的无TIMP的基质金属蛋白酶-9与分化的单核细胞及其分泌的基质金属蛋白酶-9的血管生成能力。比较将包括不同类型的髓系细胞释放/产生基质金属蛋白酶-9的动力学研究，向血管生成和肿瘤组织输送基质金属蛋白酶-9的炎性细胞的流入，以及由炎性中性粒细胞与单核细胞输送的基质金属蛋白酶-9在体内的血管生成活性的量化。目的：阐明决定中性粒细胞基质金属蛋白酶-9高促血管生成和促肿瘤进展潜能的生化机制(S)。中性粒细胞流入肿瘤组织的诱导物将被量化。无TIMP的中性粒细胞基质金属蛋白酶-9酶原的激活机制将被确定，这在体内从未被记录过。将对纤溶酶原/纤溶酶丝氨酸蛋白酶级联进行机械检测，以便在血管生成诱导过程中与激活的基质金属蛋白酶-9配对。激活的中性粒细胞基质金属蛋白酶-9的体内底物和效应分子将通过新的蛋白质组学方法进行探索。目的3：验证中性粒细胞基质金属蛋白酶-9的促血管生成活性与肿瘤通过新生血管管道的扩散有关。中性粒细胞基质金属蛋白酶-9介导的血管生成的特异性减少或刺激将直接与肿瘤细胞的血管生成水平有关，并通过活体中性粒细胞和/或纯化的中性粒细胞基质金属蛋白酶-9的体内救援实验证实。然后，结果将在一些血管生成和转移的小鼠模型中得到验证，包括原位移植和专注于类似人类疾病的TRAMP模型的转基因模型。