NETs and their modulating enzymes in age-related inflammatory diseases

NETs 及其调节酶在与年龄相关的炎症性疾病中的作用

• 批准号: 8799988
• 负责人: DENISA D WAGNER
• 金额: $ 44.04万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-12-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8799988
• 关键词: Address; Affect; Age; Aging; Animals; Biological Markers; Blood Cells; Blood Platelets; Bone Marrow; Bone Marrow Transplantation; Caloric Restriction; Cardiac; Cell Nucleus; Chimera organism; Chromatin; Chronic; Deacetylase; Disease; Disease model; Enzymes; Fibrosis; Genes; Genetic Engineering; Goals; Health; Heart; Impaired wound healing; In Vitro; Inflammation; Inflammatory; Lead; Leukocytes; Longevity; Lung; Mediating; Metabolic stress; Mitochondria; Modeling; Mus; Mutant Strains Mice; Mutation; Organ; P-Selectin; Plasma; Platelet Activation; Play; Population; Predisposition; Prevalence; Process; Processed Genes; Production; Protein-arginine deiminase; Reactive Oxygen Species; Regulation; Role; Siblings; Study models; Thrombosis; Tissues; Transgenic Mice; Wild Type Mouse; Work; age related; aging population; coronary fibrosis; extracellular; improved; in vivo; interstitial; neutrophil; overexpression; prevent; public health relevance; response

DESCRIPTION (provided by applicant): The prevalence of chronic inflammatory and thrombotic diseases is rising with the general increase in population longevity. Our application addresses the role of neutrophil extracellular traps (NETs), chromatin released by activated neutrophils, in such age-related diseases. We hypothesize that aging promotes NET formation and NETs in turn escalate chronic inflammatory and thrombotic diseases. We propose that genes regulating NETosis, such as peptidylarginine deiminase 4 (PAD4), impact aging. Our preliminary results indicate that neutrophils from old mice make more NETs and PAD4, which orchestrates NETosis, promotes cardiac fibrosis in older mice. We observed that the mitochondrial deacetylase sirtuin 3 (SIRT3), whose regulation of reactive oxygen species (ROS) is implicated in aging, is expressed by neutrophils and platelets and is a potent down-regulator of neutrophil ROS production and NETosis. We will address our hypothesis with in vitro studies and disease models in wild type (WT) and genetically engineered mutant mice. We propose three Aims: Aim 1. To study the relationship of NETosis and aging in WT mice, the effect of calorie restriction (CR) on NETosis and resulting inflammatory, thrombotic, and fibrotic responses. We will examine NETosis and the susceptibility to cardiac and lung fibrosis as a function of mouse age. We will evaluate whether DNase1, that helps clear NETs, will reduce experimental lung fibrosis. We will study the effect of CR, known to slow down aging, on NET formation and susceptibility to thrombosis and organ fibrosis. Aim 2. To investigate whether mutations in PAD4, an enzyme required for NET formation, affect aging and age-related inflammatory and thrombotic diseases. We will study the effect of PAD4- deficiency on age-related heart fibrosis and susceptibility to experimental lung fibrosis. We will follow aging healt and longevity of PAD4-/- and WT littermates. We will genetically engineer transgenic mice overproducing PAD4 in neutrophils. We anticipate that they will produce NETs more readily and have an increased propensity towards thrombosis and interstitial lung fibrosis as well as delayed wound healing. Aim 3. To evaluate the role of SIRT3 in NETosis and in disease models involving NETs. Study the function of platelet SIRT3 in platelet activation and in their priming of neutrophils towards NETosis. We are keen to evaluate the role of SIRT3 in neutrophils, platelets and in the interaction between platelets and neutrophils that may enhance NETosis. SIRT3-/- and already generated SIRT3- overexpressing mice will be evaluated in inflammation and thrombosis. Bone marrow transplant chimeric animals will be prepared to distinguish the role of blood cell versus tissue SIRT3. We hope to uncover new pharmacological targets to reduce NETs' contribution to chronic inflammatory and thrombotic processes and that this may eventually improve the health of our aging population.

描述（由申请人提供）：随着人口寿命的普遍延长，慢性炎症和血栓性疾病的患病率正在上升。我们的应用解决了中性粒细胞胞外陷阱 (NET)（活化的中性粒细胞释放的染色质）在此类与年龄相关的疾病中的作用。我们假设衰老促进 NET 的形成，而 NET 反过来又会加剧慢性炎症和血栓性疾病。我们认为调节 NETosis 的基因，例如肽基精氨酸脱亚胺酶 4 (PAD4)，会影响衰老。我们的初步结果表明，老年小鼠的中性粒细胞产生更多的 NET 和 PAD4，后者协调 NETosis，促进老年小鼠的心脏纤维化。我们观察到，线粒体脱乙酰酶 Sirtuin 3 (SIRT3) 由中性粒细胞和血小板表达，其对活性氧 (ROS) 的调节与衰老有关，并且是中性粒细胞 ROS 产生和 NETosis 的有效下调因子。我们将通过野生型（WT）和基因工程突变小鼠的体外研究和疾病模型来解决我们的假设。我们提出三个目标： 目标 1. 研究 WT 小鼠 NETosis 与衰老的关系、热量限制 (CR) 对 NETosis 以及由此产生的炎症、血栓和纤维化反应的影响。我们将检查 NETosis 以及对心脏和肺纤维化的易感性与小鼠年龄的关系。我们将评估有助于清除 NET 的 DNase1 是否会减少实验性肺纤维化。我们将研究 CR（已知可以延缓衰老）对 NET 形成以及血栓形成和器官纤维化易感性的影响。目标 2. 研究 PAD4（一种 NET 形成所需的酶）的突变是否会影响衰老以及与年龄相关的炎症和血栓性疾病。我们将研究 PAD4 缺陷对年龄相关心脏纤维化和实验性肺纤维化易感性的影响。我们将跟踪 PAD4-/- 和 WT 同窝小鼠的衰老健康状况和寿命。我们将对转基因小鼠进行基因改造，使中性粒细胞过度产生 PAD4。我们预计它们会更容易产生 NET，并且更容易发生血栓和间质性肺纤维化以及伤口愈合延迟。目标 3. 评估 SIRT3 在 NETosis 和涉及 NET 的疾病模型中的作用。研究血小板 SIRT3 在血小板激活及其启动中的功能 中性粒细胞走向 NETosis。我们热衷于评估 SIRT3 在中性粒细胞、血小板以及血小板和中性粒细胞之间可能增强 NETosis 的相互作用中的作用。 SIRT3-/-和已经生成的SIRT3-过表达小鼠将在炎症和血栓形成中进行评估。将准备骨髓移植嵌合动物来区分血细胞与组织 SIRT3 的作用。我们希望发现新的药理学靶点，以减少 NET 对慢性炎症和血栓形成过程的影响，这可能最终改善我们老龄化人口的健康。