The Monocyte/Macrophage Role in Experimental Deep Vein Thrombosis Resolution and Vein Wall Injury

摘要 血栓后综合征（PTS）是深静脉血栓形成（DVT）最常见的后遗症， 其特征在于静脉壁纤维化、瓣膜破坏和经常闭塞。据估计，这将发生在约40 - 50%的人患有DVT。没有直接的药物治疗存在，以治疗PTS，突出了最近的失败 分级压缩长袜，以防止PTS（SOX试验）。吸引试验表明，即使是那些 接受溶栓治疗的患者在2年时仍有约40%的PTS事件，而侵入性药物治疗 机械血栓清除并不能改善结果。即使采用新的治疗方法， 非维生素K拮抗剂。单核细胞/巨噬细胞（Mo/MΦ）是白细胞介导的两个关键细胞， 病理生物学过程：静脉血栓形成消退和相关的静脉壁纤维化损伤。Mo/MΦ是 根据它们的炎症或抗炎功能分类，这是体内的动态过程。为 例如，白细胞介素-1（IL-1）、IL-12分泌和细胞表面Ly 6Chi、CCR 2 ++、CX 3CR 1+抗原表达 典型地表征活化的或促炎性的Mo/MΦ。相反，IL-10的分泌、转录 Nr 4a 1因子依赖性和细胞表面Ly 6Clo、CCR 2-、CX 3CR 1 ++抗原表达的特征 交替激活Mo/MΦ，具有促愈合和抗炎活性。从出版和 初步数据显示，促炎性Ly 6Chi Mo/MΦ与早期VT有关，随后是后期向VT的过渡。 Ly6Clo。我们发现，在一个静止的小鼠VT模型中，存在促炎细胞因子环境， LyC 6 hi Mo/MΦ转变为LyC 6lo含量，并且Ly 6Clo Mo/MΦ也可以驱动VT分辨率 静脉壁纤维化损伤但Mo/MΦ在VT消退和静脉壁损伤中的作用机制尚不清楚， 以及这是否是血栓形成模型，性别和年龄依赖性尚不清楚。我们的总体假设是 VT消退和静脉壁纤维化损伤依赖于Mo/MΦ作用，依赖于模型、年龄 血栓形成静脉中Ly 6Clo Mo/MΦ的增加可改善血栓形成静脉中的Ly 6Clo Mo/MΦ。我们会解决这个问题 三个具体目标的假设。具体目标1：确定局部环境和细胞因素， 在血栓形成静脉中驱动Ly 6Chi和Ly 6Clo Mo/MΦ表型，与性别、年龄和血栓形成模型相关 变化量具体目标2：直接确定Mo/MΦ介导的VT消退和静脉血栓形成的机制。 墙伤。具体目标3：确定血栓环境中的目标Mo/MΦ极化是否可以 促进VT消退和静脉壁愈合。小鼠室速模型，年龄和性别不同，Mo/MΦ 条件性缺失和转基因小鼠以及分子、免疫学和体内成像将用于 实现这些目标。本文提出的实验将通过定义 Mo/MΦ介导的VT消退和静脉壁损伤的机制，并将测试新的 评估和药剂以增加促愈合Mo/MΦ活性，并可能转化为PTS疗法。

ABSTRACT Post-thrombotic syndrome (PTS) is the most common sequelae from deep vein thrombosis (DVT), characterized by vein wall fibrosis, valve destruction, and often occlusion. It is estimated to occur in ~40 – 50% of those suffering a DVT. No direct medical therapy exists to treat PTS, highlighted by the recent failure of graded compression stockings to prevent PTS (SOX trial). The ATTRACT trial suggested that even those patients treated with thrombolysis still have a ~40% incident PTS at 2 years, and invasive pharmaco- mechanical thrombus removal did not improve outcomes. Significant bleeding risks remain even with the new non-vitamin-K antagonists. Monocyte/macrophages (Mo/MΦ) are the primary leukocyte directing two key pathobiologic processes: venous thrombosis resolution and the associated vein wall fibrotic injury. Mo/MΦ are classified by their inflammatory or anti-inflammatory functions, which is a dynamic process in vivo. For example, interleukin-1 (IL-1), IL-12 secreting and cell surface Ly6Chi, CCR2++, CX3CR1+ antigen expression characterizes classically activated, or pro-inflammatory Mo/MΦ. Conversely, IL-10 secreting, transcription factor Nr4a1 dependent, and cell surface Ly6Clo, CCR2-, CX3CR1++ antigen expression characterizes alternatively activated Mo/MΦ with pro-healing and inflammation resolving activities. From published and preliminary data, pro-inflammatory Ly6Chi Mo/MΦ are involved with early VT, followed by a later transition to Ly6Clo. We show that in a stasis murine model of VT that a pro-inflammatory cytokine milieu exists, that early LyC6hi Mo/MΦ changes over to a LyC6lo content, and that Ly6Clo Mo/MΦ may drive both VT resolution as well as vein wall fibrotic injury. However, the mechanism of Mo/MΦ actions in VT resolution and vein wall injury, as well as whether this is thrombogenic model, sex and age dependent is not known. Our overall hypothesis is that VT resolution and vein wall fibrotic injury is dependent on Mo/MΦ actions, is dependent on model, age, and sex, and can be ameliorated with increasing Ly6Clo Mo/MΦ in the thrombosed vein. We will address this hypothesis by three specific aims. Specific Aim 1: To define the local environmental and cellular factors that drive Ly6Chi and Ly6Clo Mo/MΦ phenotypes in the thrombosed vein, with sex, age, and thrombogenic model variation. Specific Aim 2: To directly determine the Mo/MΦ mediated mechanisms of VT resolution and vein wall injury. Specific Aim 3: To determine if targeted Mo/MΦ polarization within the thrombus environment can promote VT resolution and vein wall healing. Murine models of VT, with variations of age and sex, Mo/MΦ conditional deleted and transgenic mice and molecular, immunological, and in vivo imaging will be used to accomplish these aims. The proposed experiments herein will significantly move the field forward by defining the mechanisms of Mo/MΦ mediated actions in VT resolution and vein wall injury, and will test novel assessments and agents to increase pro-healing Mo/MΦ activity with potentially translation to PTS therapies.