Neutrophil Extracellular Traps and Host Immunity

项目摘要 除了经典的吞噬和细胞内氧化杀灭病原体的模式外，最近还有一种 已发现的中性粒细胞的抗菌功能是形成细胞外陷阱(中性粒细胞外 陷阱，蚊帐)，它可以捕捉和杀死病原体细胞外。AS网络形成(NETsis)通常需要 活性氧物种(ROS)的产生，我们和其他人发现，中性粒细胞从慢性 肉芽肿病患者和NADPH氧化酶复合体突变的Gp91Phox/-CGD小鼠 在体外和体内对各种刺激和肺部细菌的反应表现出网络生成受损 感染。我们最近报道了FDA批准的选择性雌激素受体(ER)他莫昔芬(TMX) 治疗乳腺癌的改良剂，在不依赖ROS的CGD中性粒细胞中诱导抗菌网 举止。我们进一步证明自噬的激活是诱导TMX介导的必要条件和充分条件。 篮网。除了这份开创性的报告外，拟议研究的前提是来自我们的初步 数据表明TMX通过非典型自噬的ROS和ER非依赖性NETsis的新途径 激活。这两个拟议的具体目标将使TMX成为具有抗菌和抗病毒作用的网络诱导剂 临床前期小鼠CGD和人CGD中性粒细胞的炎症效应(AIM 1)；并阐明TMX介导的 中性粒细胞中的非典型自噬信号通路，最终导致核层解体 协助放行净网人员(目标2)。我们的研究为一种新的自噬现象提供了重要的机械论见解 TMX激活的途径，不仅对网络研究，而且对开发 自噬和网络治疗传染病和自身免疫性疾病。通过利用井中的中性粒细胞- 这些研究是NIH临床中心CGD患者的特征，也提供了一个令人兴奋的机会 用于临床前测试CGD中的TMX以恢复中性粒细胞的抗菌功能以对抗肺炎 细菌感染，常见于这些患者。

Project Summary In addition to the classical mode of phagocytosis and intracellular oxidative killing of pathogens, a recently discovered antimicrobial function of neutrophils is the formation of extracellular traps (Neutrophil Extracellular Traps, NETs), which can trap and kill pathogens extracellularly. As NET formation (NETosis) generally requires reactive oxygen species (ROS) generation, we and others have found that neutrophils from Chronic Granulomatous Diseases (CGD) patients and Gp91phox-/- CGD mice with mutations in NADPH oxidase complex exhibit impaired NET generation in-vitro and in-vivo in response to various stimuli and pulmonary bacterial infection. We recently reported that Tamoxifen (TMX), an FDA approved selective estrogen receptor (ER) modifier for treatment of breast cancer, induces antimicrobial NETs in CGD neutrophils in a ROS independent manner. We further showed that activation of autophagy is necessary and sufficient to induce TMX-mediated NETs. In addition to this seminal report, the premise of the proposed research is derived from our preliminary data indicating a novel pathway of ROS-and ER-independent NETosis by TMX via a non-canonical autophagy activation. The two proposed specific aims will establish TMX as NET-inducing agent with antimicrobial and anti- inflammatory effect in preclinical murine CGD and human CGD neutrophils (Aim 1); and elucidate TMX-mediated non-canonical autophagy signaling pathway in neutrophils that culminates in disintegration of nuclear lamina to facilitate the release of NETs (Aim 2). Our studies provide important mechanistic insights into a novel autophagy pathway activated by TMX which will have implications not only for NET research but also for exploiting autophagy and NETs to treat infectious and autoimmune diseases. By leveraging neutrophils from a well- characterized cohort of CGD patients at NIH Clinical Center, these studies also present an exciting opportunity for preclinical testing of TMX in CGD to restore antimicrobial function of neutrophils to combat pneumonic bacterial infections, frequently observed in these patients.