NETS in thrombosis and inflammatory responses

 描述(申请人提供)：当中性粒细胞激活时，肽基精氨酸脱亚胺酶4(PAD4)可能转移到细胞核中形成瓜氨酸组蛋白。这会使染色质解聚，染色质以中性粒细胞外陷阱(Net)的形式释放。蚊帐捕获微生物，但也促进炎症。与这一应用相关的是，PAD4/Nets在病理性血栓形成中起着重要作用，癌症可以刺激它们的形成。这一提议的中心假设是：Net参与了稳定的、有组织的和有血管的血栓的形成，并且Net的破裂是溶栓所必需的。血栓形成促进邻近血管壁和远处器官的净沉积，导致血栓后综合征(PTS)和全身促凝血和促炎状态的增加。最后，PAD4/NETs提高了肿瘤的凝固性，促进了肿瘤的血管生成和生长。为了解决这些假说，我们将研究野生型(WT)和基因工程小鼠的几种深静脉血栓形成(DVT)和乳腺癌模型。将要使用的特殊基因敲除包括PAD4-/-和新生成的DNase1-/-小鼠。新技术将被应用于提取新生血管的3D模型，以从血栓生成血管计量学数据。此外，还将测试剪切激活纳米疗法(SA-NTS)以诱导血栓溶解。该提案有两个具体目标：目的1：研究NETS在静脉血栓形成和溶栓中的作用。为此，我们建议研究NETs在深静脉血栓形成的不同阶段以及由此导致的血管壁纤维化中的作用。我们将评估细胞外染色质和血小板在血栓新生血管形成中的作用，以及染色质和新生血管如何影响血栓溶解。目的：研究Nets/PAD4在血栓诱导的全身炎症和癌症诱导的血栓形成中的作用。在这个目标中，我们将检查有DVT经历的小鼠的全身效应和易感性，以及Net在多大程度上有助于癌症的促凝和促血管生成活动。血栓形成，包括深静脉血栓/肺栓塞，现在是美国最大的杀手。我们希望我们的研究将扩大对NETs在血栓形成中所起作用的理解，并有助于指导新的治疗和预防病理性血栓形成、PTS和癌症诱导的血栓形成。

 DESCRIPTION (provided by applicant): Upon neutrophil activation, peptidylarginine deiminase 4 (PAD4) may translocate to the nucleus to citrullinate histones. This decondenses chromatin which is released as neutrophil extracellular traps (NETs). NETs trap microbes but also promote inflammation. Relevant to this application, PAD4/NETs play an important role in pathological thrombosis and their formation can be stimulated by cancer. The central hypotheses of this proposal are: NETs are involved in the formation of a stable organized and vascularized thrombus and breaking up NETs is necessary for thrombolysis. Thrombosis promotes NET deposition in the adjacent vessel wall and in distant organs leading to post-thrombotic syndrome (PTS) and an increased systemic pro-coagulant and pro-inflammatory state. Finally, PAD4/NETs elevate coagulation in cancer and promote tumor angiogenesis and growth. To address the hypotheses, we will study several models of deep vein thrombosis (DVT) and breast cancer in wild-type (WT) and genetically engineered mice. The special knockouts to be used include PAD4-/- and newly generated DNase 1-/- mice. Novel technologies will be applied to extract 3D models of the neovasculature to generate vasometrics from the thrombi. Also, shear-activated nanotherapeutics (SA-NTs) will be tested to induce thrombolysis. The proposal has two specific aims: Aim1: to study NETs in venous thrombosis and thrombolysis. In this aim, we propose to investigate the role of NETs in various stages of DVT formation and in resulting fibrosis of the vessel wall. We will evaluate the role of extracellular chromatin and platelets in thrombus neo- vascularization and how chromatin and neovascularization affect thrombolysis. Aim 2: to study NETs/PAD4 in thrombosis-induced systemic inflammation and cancer-induced thrombosis. In this aim, we will examine what systemic effects and susceptibilities a mouse with DVT experiences and to what extent NETs contribute to pro-coagulant and pro-angiogenic activities of cancer. Thrombosis, including DVT/pulmonary embolism, is now the biggest killer in United States. We hope that our studies will broaden understanding of the roles NETs play in thrombosis and help to guide new treatments and prevention of pathological thrombosis, PTS and cancer-induced thrombosis.