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: 8690794
• 负责人: JAMES P QUIGLEY
• 金额: $ 38.14万
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8690794
• 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（由申请人提供）：恶性进展的早期步骤之一是肿瘤血管生成，为正在发育的肿瘤提供营养，并可能为肿瘤细胞逃逸提供途径。在许多移植和转基因模型中，BMDC 亚群被证明可传递 MMP-9 以介导血管生成开关，并且通常，传递 MMP-9 的 BMDC 被鉴定为单核细胞/巨噬细胞。一些研究表明，中性粒细胞可能作为另一种 BMDC 类型，因为它们也表达 MMP-9，并且与肿瘤发展相关（尽管是短暂的）。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 proMMP-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浸润肿瘤间质组织并快速释放其预存的、高度血管生成的、不含TIMP的MMP-9，我们将挑战现有的范例，即单核细胞样细胞是血管生成开关的唯一诱导剂。我们提出一个总体假设，即中性粒细胞及其独特的 MMP-9 的初始递送是肿瘤进展中关键且可靶向的早期事件。为了检验这一假设，我们提出以下具体目标： 目标 1：直接比较炎症性中性粒细胞及其独特的无 TIMP MMP-9 的血管生成能力与分化单核细胞及其分泌的 MMP-9。比较将包括不同骨髓细胞类型的 MMP-9 释放/产生的动力学研究、递送 MMP-9 的炎症细胞流入血管生成和肿瘤组织，以及量化炎症中性粒细胞与单核细胞递送的 MMP-9 的体内血管生成活性。目标 2：阐明决定中性粒细胞 MMP-9 的高促血管生成和促肿瘤进展潜力的生化机制。中性粒细胞流入肿瘤组织的诱导物将被量化。将确定无 TIMP 的中性粒细胞 MMP-9 酶原的激活机制，该机制在体内从未有过记录。将在机制上检查纤溶酶原/纤溶酶丝氨酸蛋白酶级联在血管生成诱导期间与激活的 MMP-9 的配合。激活的中性粒细胞 MMP-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测。目标 3：验证中性粒细胞 MMP-9 的促血管生成活性与通过新血管导管的肿瘤播散存在因果关系。中性粒细胞MMP-9介导的血管生成的特异性减少或刺激将与肿瘤细胞内渗的水平直接相关，并通过使用活中性粒细胞和/或纯化的中性粒细胞MMP-9的体内拯救实验得到证实。然后，结果将在许多血管生成和转移的小鼠模型中得到验证，包括原位移植和侧重于人类疾病样 TRAMP 模型的转基因模型。