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也在上升。我们的应用程序解决了中性粒细胞胞外陷阱（NET），染色质释放的激活中性粒细胞，在这种年龄相关性疾病的作用。我们推测，老化促进NET的形成和NET反过来升级慢性炎症和血栓性疾病。我们认为NETosis的调控基因，如肽基精氨酸脱亚胺酶4（PAD 4），影响衰老。我们的初步结果表明，来自老年小鼠的中性粒细胞产生更多的NETs，而协调NETosis的PAD 4促进老年小鼠的心脏纤维化。我们观察到，线粒体去乙酰化酶sirtuin 3（SIRT 3），其调节活性氧（ROS）参与衰老，是由中性粒细胞和血小板表达，是一种有效的下调中性粒细胞ROS的生产和NETosis。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小鼠中NETosis与衰老的关系，卡路里限制（CR）对NETosis的影响以及导致的炎症、血栓形成和纤维化反应。我们将研究NETosis和心脏和肺纤维化的易感性作为小鼠年龄的函数。我们将评估有助于清除NETs的DNase 1是否会减少实验性肺纤维化。我们将研究已知可以延缓衰老的CR对NET形成以及对血栓形成和器官纤维化的易感性的影响。目标2.研究PAD 4（NET形成所需的酶）突变是否影响衰老和年龄相关的炎症性和血栓性疾病。我们将研究PAD 4缺乏对年龄相关性心脏纤维化和实验性肺纤维化易感性的影响。我们将跟踪PAD 4-/-和WT同窝仔的衰老、健康和寿命。我们将对转基因小鼠进行基因工程改造，使其在中性粒细胞中过度产生PAD 4。我们预计，它们将更容易产生NET，并增加血栓形成和间质性肺纤维化以及延迟伤口愈合的倾向。目标3。评估SIRT 3在NETosis和涉及NET的疾病模型中的作用。研究血小板SIRT 3在血小板活化及其启动中的作用。 嗜中性粒细胞向NETosis。我们热衷于评估SIRT 3在中性粒细胞、血小板中的作用，以及在可能增强NETosis的血小板和中性粒细胞之间的相互作用中的作用。将在炎症和血栓形成中评价SIRT 3-/-和已经产生的SIRT 3过表达小鼠。将制备骨髓移植嵌合动物以区分血细胞与组织SIRT 3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贡献，这可能最终改善我们老龄化人口的健康。