NETs in the development of lupus and its cardiovascular complications

NETs 在狼疮及其心血管并发症发生发展中的作用

• 批准号: 8853182
• 负责人: Daniel T Eitzman
• 金额: $ 39.56万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8853182
• 关键词: Affect; Amidines; Antigen-Antibody Complex; Atherosclerosis; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Blood Platelets; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Carotid Artery Thrombosis; Carotid Atherosclerotic Disease; Chemicals; Chromatin; Chromatin Fiber; Complex; Coronary; Data; Dendritic Cells; Development; Disease; Endothelial Cells; Enzymes; Event; Health; Histones; Human; Individual; Inflammation; Inflammatory; Interferons; Kidney; Knock-out; Knowledge; Lead; Link; Lupus; Mediator of activation protein; Medical; Michigan; Modeling; Molecular; Mus; Nuclear; Organ; Ornithine; Pathogenesis; Patients; Pharmaceutical Preparations; Process; Production; Role; Skin; Skin Manifestations; Source; Stimulus; Support Groups; Surrogate Markers; Systemic Lupus Erythematosus; Therapeutic; Therapeutic Intervention; Vasodilation; Venous Thrombosis; Work; adaptive immunity; antimicrobial peptide; atherogenesis; cardiovascular risk factor; cell injury; cohort; density; design; endothelial dysfunction; extracellular; granulocyte; in vivo; inhibitor/antagonist; neutrophil; novel; premature; programs; response

DESCRIPTION (provided by applicant): Recent work from our group and others implicate aberrant neutrophil extracellular trap (NET) formation, the programmed release of chromatin fibers in complex with antimicrobial peptides, as a potentially crucial phenomenon involved in the induction of autoimmune responses, organ damage and premature cardiovascular disease (CVD) in systemic lupus erythematosus (SLE). Our data are supported by recent studies also linking NETosis to idiopathic atherosclerosis and to venous thrombosis. This competing renewal application will determine the in vivo importance of aberrant NETosis in CV damage and other manifestations of SLE, with obvious implications for medical therapy in patients affected by this condition. Aim 1: Explore the impact of NET inhibition on murine models of SLE vascular damage. While molecular events implicated in NET formation are incompletely understood, recent evidence indicates that histone citrullination is crucial for chromatin decondensation leading to NETosis6. Neutrophils express high levels of peptidylarginne deiminase-4 (PAD4), the enzyme that catalyzes histone citrullination7. Mice that lack PAD4 have decreased NETosis upon stimulation8. These observations indicate that PAD4 is crucial for NETosis and could be explored as a target in SLE and its associated vascular complications. Our collaborator, Dr. Paul Thompson (Scripps), has developed a novel PAD inhibitor N-�-benzoyl-N5-(2-chloro-1-iminoethyl-L-ornithine amide (Cl-amidine) that inhibits nuclear PAD4 with low �M potency9,10. Cl-amidine inhibits NET formation in human and murine SLE neutrophils (see preliminary data) and has previously been well-tolerated in other murine inflammatory models. We will utilize chemical PAD4 inhibition and available PAD4 knockout (KO) models to block NETosis in murine SLE and in models of lupus with accelerated atherogenesis, allowing us, to assess the in vivo impact of NET inhibition on models of lupus-associated vascular damage: carotid thrombosis, neoangiogenesis, endothelial-dependent vasorelaxation and atherosclerosis. We predict that blocking NET formation will significantly reduce vascular damage in SLE. Aim 2: Determine if other SLE manifestations can be abrogated by NET inhibition. Recent data from various groups supports the concept that NETosis is a stimulus for IFN-� production, a phenomenon that may promote autoimmunity3,13,14; further, there are emerging hints that IFN-� may stimulate NETosis, thereby resulting in a harmful, self-perpetuating cycle. As NETs can damage the vasculature directly, Aim 1 is a logical place to begin our assessment. However, we anticipate that NET inhibition will also have a global impact on SLE disease activity and on type I IFN responses. We will assess the role of aberrant NETosis in autoantibody (autoAb) synthesis, induction of type I IFN-responses, adaptive immune responses and organ inflammation and damage in murine SLE, with a particular focus on kidney and skin damage. Aim 3: Determine the association of aberrant neutrophils, NETs and anti-neutrophil Abs in surrogate markers of endothelial dysfunction and CV risk in SLE patients. The association of neutrophil:platelet aggregates (NPAs), low-density granulocytes (LDGs), NETosis and autoAbs to neutrophil-derived immunostimulatory products, with endothelial dysfunction, subclinical atherosclerosis (carotid and coronary), type I IFN signatures and surrogate markers of vascular damage will be determined in cross-sectional and longitudinal analyses in a well-characterized CV Lupus Cohort established at the U. of Michigan. Results from this proposal will provide crucial information regarding the role of neutrophils and aberrant NETosis in the pathogenesis of autoimmune responses and organ damage in SLE, and may lead to changes in the therapeutic armamentarium in individuals with this devastating condition. If we confirm our hypothesis and extend the knowledge regarding mechanisms of effect in the studies proposed herein, Cl-amidine or other PAD-4 inhibitors may represent the first of a novel class of disease-modifying drugs for lupus that target a recently described process which may be crucial in autoimmune responses and organ damage in SLE.

描述(由申请人提供)： 我们和其他人最近的工作表明，异常的中性粒细胞细胞外陷阱(Net)的形成，即染色质纤维在与抗菌肽的复合体中的程序性释放，是系统性红斑狼疮(SLE)诱导自身免疫反应、器官损伤和过早心血管疾病(CVD)的潜在关键现象。我们的数据也得到了最近研究的支持，这些研究还将NETsis与特发性动脉粥样硬化和静脉血栓联系起来。这一竞争性的更新应用将确定变异的NETsis在心血管损害和SLE的其他表现中的体内重要性，对受这种情况影响的患者的药物治疗具有明显的意义。目的1：探讨净抑制对系统性红斑狼疮小鼠血管损伤模型的影响。虽然与网络形成有关的分子事件还不完全清楚，但最近的证据表明，组蛋白瓜氨酸化是染色质解缩导致NETosis的关键。中性粒细胞表达高水平的多肽精氨酸脱亚胺酶-4(PAD4)，这种酶催化组蛋白瓜氨酸化7。缺乏PAD4的小鼠在刺激后减少了NETase 8。这些观察表明，PAD4在NETsis中是至关重要的，可以作为SLE及其相关血管并发症的靶点。我们的合作者保罗·汤普森博士(斯克里普斯)已经开发出一种新型的PAD抑制剂N-�-benzoyl-N5-(2-chloro-1-iminoethyl-L-ornithine酰胺(氯-酰胺)，它以低�M值9，10抑制核PAD4。CL-嘧啶抑制人和小鼠系统性红斑狼疮中性粒细胞的净形成(见初步数据)，并且以前在其他小鼠炎症模型中具有良好的耐受性。我们将利用化学PAD4抑制和现有的PAD4基因敲除(KO)模型来阻断小鼠SLE和加速动脉粥样硬化的狼疮模型中的NETsis，使我们能够在体内评估Net抑制对狼疮相关血管损伤模型的影响：颈动脉血栓形成、新生血管生成、内皮依赖的血管松弛和动脉粥样硬化。我们预测，阻断网络的形成将显著减少SLE的血管损害。目的2：确定其他系统性红斑狼疮的表现是否可以通过网络抑制来消除。来自不同团体的最新数据支持这一概念，即干扰素-�是一种刺激产生干扰素-�的现象，这一现象可能会促进自身免疫3，13，14；此外，有新的迹象表明，干扰素-DNA可能会刺激网纹病，从而导致有害的、自我持续的循环。由于NETS可以直接损害血管系统，因此目标1是我们开始评估的合乎逻辑的地方。然而，我们预计净抑制也将对SLE疾病活动和I型干扰素反应产生全球影响。我们将评估异常网状病毒在小鼠系统性红斑狼疮自身抗体(AutoAb)合成、诱导I型干扰素反应、适应性免疫反应以及器官炎症和损伤中的作用，尤其是肾脏和皮肤损伤。目的：探讨中性粒细胞、Net和抗中性粒细胞抗体异常与系统性红斑狼疮患者血管内皮细胞功能障碍和心血管风险的关系。中性粒细胞：血小板聚集(NPA)、低密度粒细胞(LDGs)、网织红细胞增多和自身抗体与中性粒细胞衍生的免疫刺激产物、内皮功能障碍、亚临床动脉粥样硬化(颈动脉和冠状动脉)、I型干扰素签名和血管损伤的替代标记物的相关性将在密歇根大学建立的具有良好特征的CV狼疮队列中通过横断面和纵向分析来确定。这项建议的结果将提供有关中性粒细胞和异常网织红细胞在系统性红斑狼疮自身免疫反应和器官损伤的发病机制中所起作用的关键信息，并可能导致患有这种毁灭性疾病的个体的治疗设施的变化。如果我们证实我们的假设，并扩展在本文提出的研究中关于作用机制的知识，氯-嘧啶或其他PAD-4抑制剂可能代表第一类针对最近描述的过程的狼疮新的疾病修饰药物，该过程可能在SLE的自身免疫反应和器官损伤中至关重要。