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: 8294290
• 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 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8294290
• 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; Image; 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; 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. PUBLIC HEALTH RELEVANCE: Deaths from cancer occur mainly because tumor cells spread from the primary tumor site to other vital organs and tissues through the new angiogenic vasculature of the primary tumor. One of the non-tumor cell types of the host that can actively contribute to the induction of new angiogenic blood vessels feeding the tumor is the inflammatory neutrophil, which can import molecules that directly induce the formation of new blood vessels and do so early on in tumor progression. The goal of the proposed research is to biochemically and mechanistically examine the pro-malignant role of the unique neutrophil enzyme, MMP-9, one of the few molecules released by neutrophils that is a potent inducer of angiogenesis and likely an inducer of tumor metastasis and thus could be therapeutically targeted in cancers associated with inflammatory neutrophil infiltration.

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，为发育中的肿瘤提供营养，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，显示BMDC的亚群递送MMP-9以介导血管生成转换，并且通常，MMP-9递送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一种替代的BMDC类型，因为它们也表达MMP-9，并已与肿瘤的发展，虽然短暂。然而，炎症性中性粒细胞，其唯一明确的作用，在急性感染和损伤，通常被忽视的贡献者肿瘤进展。我们最近证明，中性粒细胞产生一种独特的，无TIMP形式的proMMP-9，使其成为一种有效的血管生成剂。鉴于这一证据以及嗜中性粒细胞浸润肿瘤间质组织并迅速释放其预储存的、高度血管生成的无TIMP的MMP-9，我们将挑战现有的范式，即单核细胞样细胞是血管生成开关的唯一诱导物。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的关键和可靶向的早期事件。目的1：直接比较炎性中性粒细胞及其独特的无TIMP的MMP-9与分化的单核细胞及其分泌的MMP-9的血管生成能力。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，MMP-9递送炎性细胞流入血管生成和肿瘤组织，以及定量炎性中性粒细胞相对于单核细胞递送的MMP-9的体内血管生成活性。目的2：阐明决定神经元MMP-9的高促血管生成和促肿瘤进展潜力的生化机制。将对中性粒细胞流入肿瘤组织的诱导物进行定量。将确定无TIMP的中性粒细胞MMP-9酶原的激活机制，这在体内从未有过记录。纤溶酶原/纤溶酶丝氨酸蛋白酶级联将在血管生成诱导过程中与活化的MMP-9合作进行机械检查。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。目的3：证实神经元MMP-9的促血管生成活性与肿瘤通过新生血管管道扩散的因果关系。嗜中性粒细胞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，并通过使用活的嗜中性粒细胞和/或纯化的嗜中性粒细胞MMP-9的体内拯救实验证实。然后将在许多血管生成和转移的鼠模型中验证结果，包括原位移植和转基因模型，重点是人类疾病样TRAMP模型。 公共卫生相关性：癌症导致的死亡主要是因为肿瘤细胞从原发肿瘤部位通过原发肿瘤的新生血管系统扩散到其他重要器官和组织。可以积极促进诱导滋养肿瘤的新血管生成血管的宿主的非肿瘤细胞类型之一是炎性中性粒细胞，其可以输入直接诱导新血管形成的分子，并且在肿瘤进展的早期这样做。拟议研究的目标是从生物化学和机制上研究独特的中性粒细胞酶MMP-9的促恶性作用，MMP-9是中性粒细胞释放的少数分子之一，是血管生成的有效诱导剂，可能是肿瘤转移的诱导剂，因此可以在与炎症性中性粒细胞浸润相关的癌症中进行治疗靶向。