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: 8504062
• 负责人: Daniel T Eitzman
• 金额: $ 39.55万
• 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 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8504062
• 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; Functional disorder; Histones; Human; Immune response; 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; antimicrobial peptide; atherogenesis; cardiovascular risk factor; cell injury; cohort; density; design; extracellular; granulocyte; in vivo; inhibitor/antagonist; neutrophil; novel; premature; programs; public health relevance; 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）： 我们小组和其他人最近的工作涉及异常中性粒细胞胞外陷阱（NET）的形成，染色质纤维与抗菌肽复合物的程序性释放，作为一个潜在的关键现象，参与诱导系统性红斑狼疮（SLE）的自身免疫反应，器官损伤和过早的心血管疾病（CVD）。我们的数据得到了最近研究的支持，这些研究也将NETosis与特发性动脉粥样硬化和静脉血栓形成联系起来。这种竞争性的更新申请将确定异常NETosis在CV损伤和SLE其他表现中的体内重要性，对受这种疾病影响的患者的药物治疗具有明显的意义。目的1：探讨NET抑制剂对SLE血管损伤小鼠模型的影响。虽然NET形成中涉及的分子事件尚未完全了解，但最近的证据表明，组蛋白瓜氨酸对导致NETosis的染色质去浓缩至关重要6。中性粒细胞表达高水平的肽基精氨酸脱亚胺酶-4（PAD 4），该酶催化组蛋白瓜氨酸化7。缺乏PAD 4的小鼠在刺激后NETosis减少8。这些观察结果表明，PAD 4对NETosis至关重要，可以作为SLE及其相关血管并发症的靶点进行探索。我们的合作者Paul Thompson博士（Scripps）开发了一种新型PAD抑制剂N-<$-苯甲酰基-N5-（2-氯-1-亚氨基乙基-L-鸟氨酸酰胺（Cl-脒），可抑制低<$M效力的核PAD 4 9，10。Cl-脒抑制人和鼠SLE中性粒细胞中的NET形成（见初步数据），并且先前在其它鼠炎症模型中耐受良好。 我们将利用化学PAD 4抑制和可用的PAD 4敲除（KO）模型来阻断小鼠SLE和具有加速动脉粥样硬化形成的狼疮模型中的NET，使我们能够评估NET抑制对狼疮相关血管损伤模型的体内影响：颈动脉血栓形成、新血管生成、内皮依赖性血管舒张和动脉粥样硬化。我们预测，阻止NET的形成将显着减少SLE的血管损伤。目的2：确定NET抑制是否可以消除其他SLE表现。来自不同团体的最新数据支持这一概念，即NETosis是刺激IFN-<$生产的一种现象，可能会促进自身免疫3，13，14;此外，有迹象表明IFN-<$可能会刺激NETosis，从而导致有害的，自我延续的循环。由于NET可直接损伤血管系统，因此目标1是我们评估的开始位置。 然而，我们预计NET抑制也将对SLE疾病活动和I型IFN应答产生全球性影响。我们将评估异常NETosis在自身抗体（autoAb）合成，诱导I型IFN-应答，适应性免疫应答以及小鼠SLE中器官炎症和损伤中的作用，特别关注肾脏和皮肤损伤。目标3：确定异常中性粒细胞、NET和抗中性粒细胞Ab在SLE患者内皮功能障碍和CV风险的替代标志物中的相关性。 中性粒细胞：血小板聚集体（NPAs）、低密度粒细胞（LDGs）、NETosis和自体抗体与嗜中性粒细胞衍生的免疫刺激产物的相关性，以及内皮功能障碍、亚临床动脉粥样硬化（颈动脉和冠状动脉）、I型IFN信号和血管损伤的替代标志物的关系将在美国建立的一个充分表征的CV狼疮队列的横断面和纵向分析中确定。密歇根州。从这一建议的结果将提供关键信息的作用，中性粒细胞和异常NETosis的自身免疫反应和器官损伤的发病机制在SLE，并可能导致在个人与这种破坏性的条件下的治疗设备的变化。如果我们证实了我们的假设，并扩展了本文提出的研究中有关作用机制的知识，Cl-脒或其他PAD-4抑制剂可能代表了一类新型狼疮疾病修饰药物中的第一种，这些药物靶向最近描述的过程，该过程可能在SLE的自身免疫反应和器官损伤中至关重要。