The Role of IL-1R1 in cell transitions in Aortic Aneurysm Pathogenesis

IL-1R1 在主动脉瘤发病机制中细胞转变中的作用

• 批准号: 9042424
• 负责人: Gorav Ailawadi
• 金额: $ 48.66万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9042424
• 关键词: Abdominal Aortic Aneurysm; Affect; Aneurysm; Angiotensin II; Antibodies; Aortic Aneurysm; Apoptosis; Attenuated; B-Lymphocytes; Blood; Blood Circulation; CCL2 gene; Caliber; Cardiovascular Diseases; Cause of Death; Cell Culture System; Cell Lineage; Cells; Chronic; Clinical; Clinical Trials; Collagen; Coupled; Data; Development; Disease; Elastases; Elastin; Elderly; Epigenetic Process; Exhibits; Flow Cytometry; Gelatinase A; Gelatinase B; Genes; Genetic; Goals; Harvest; Health; Histology; Human; IL1R1 gene; Immune; Immunohistochemistry; Incidence; Infiltration; Inflammation; Inflammatory; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Intervention; Knock-out; Knockout Mice; Label; Laboratories; Lesion; Leukocytes; Measures; Medical; Methods; Microscopic; Modeling; Mus; Muscle; Myelogenous; Myeloid Cells; Myocardial Infarction; Myosin Heavy Chains; Neutrophil Infiltration; Pathogenesis; Pathology; Pathway interactions; Patients; Perfusion; Phenotype; Plasminogen Activator Inhibitor 1; Play; Prevalence; Prevention; Production; Publications; Publishing; Pump; RANTES; Recombinant Interleukin-1; Resolution; Role; Rupture; Sampling; Secondary to; Serine Protease; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Staging; T-Lymphocyte; TNF gene; Tamoxifen; Testing; Tissues; Translating; Western Blotting; anakinra; attenuation; base; cell type; clinically relevant; cytokine; inhibitor/antagonist; macrophage; male; mouse model; neutrophil; novel; novel marker; prevent; protective effect; receptor; treatment strategy

 DESCRIPTION (provided by applicant): Hypothesis: Our previous studies using elastase-perfusion models of abdominal aortic aneurysms (AAA) have shown that IL-1R1 KO mice have a decreased incidence and size of aneurysm formation. We have also recently shown that treatment with the IL-1 pathway inhibitor, anakinra, in both prevention and treatment studies results in significant protection from aneurysm formation in a mouse model of AAA. In the current proposal, we will investigate the role of IL-1 signaling in smooth muscle and macrophage phenotypic transitions using novel rigorous lineage tracing mouse models of abdominal aortic aneurysm (AAA) formation and to examine the mechanism of IL-1R1 signaling in AAA formation as a method to selectively determine treatment strategies to protect against AAA formation. Methods: We will use a elastase perfusion and Angiotensin II murine models of AAA coupled with rigorous smooth muscle and macrophage lineage tracing analysis in conditional IL-1 receptor 1 KO mice. Anakinra treatment will be administered via osmotic pump to determine the role of Anakinra treatment on smooth muscle cell and macrophage transitions during aneurysm formation. Aortic diameter will be measured on day 3, 7, 14, and 28. Aortic tissue will be harvested to analyze pro-inflammatory cytokine (IL-1β, TNF-α, MCP-1, IL-6, and RANTES), MMP-2 and MMP-9 activity by zymography, serine proteases (uPA, tPA and PAI-1) by western blots, elastin and collagen degradation as well as aortic smooth muscle expression by histology, and immune cell (macrophages, T cells, neutrophils, B cells) infiltration by flow cytometry. Results: Preliminary data found significant attenuation in aneurysm formation in IL-1R1 KO mice 14 days following elastase perfusion in AAA formation. Wild-type male mice administered anakinra via osmotic pump 3 days before TAA formation also exhibited decreased aortic diameter. AAA formation in IL1β and IL1R1 KO male mice display a decreased aortic diameter, decreased cytokine production (TNFα, MCP-1, IL-6 and IL-27), decreased MMP-9 expression and decreased macrophage and neutrophil infiltration compared to wild-type male mice. Finally, IL-1R1 is increased in human AAA samples versus non-aneurysmal controls via qPCR and immunohistochemistry. Conclusions: IL-1 pathway inhibition can attenuate aneurysm formation and inflammation in the topical elastase murine model of AAA. We propose to delineate the mechanism of IL-1 signaling in AAA formation using novel lineage tracing analysis in smooth muscle cell and macrophages and to examine the role of IL-1 signaling in the pathology of aneurysm formation with the overall goal to develop aneurysm treatment.

 描述（由申请人提供）：假设：我们先前使用腹主动脉瘤（AAA）弹性蛋白酶灌注模型的研究表明，IL-1 R1 KO小鼠的动脉瘤形成发生率和大小降低。我们最近还表明，在预防和治疗研究中，用IL-1通路抑制剂阿那白滞素治疗可显著保护AAA小鼠模型免于动脉瘤形成。在目前的提案中，我们将使用腹主动脉瘤（AAA）形成的新的严格谱系追踪小鼠模型研究IL-1信号在平滑肌和巨噬细胞表型转变中的作用，并研究IL-1 R1信号在AAA形成中的机制，作为选择性确定治疗策略以防止AAA形成的方法。研究方法：我们将在条件性IL-1受体1 KO小鼠中使用弹性蛋白酶灌注和血管紧张素II AAA小鼠模型，并结合严格的平滑肌和巨噬细胞谱系追踪分析。阿那白滞素治疗将通过渗透泵给药，以确定阿那白滞素治疗对动脉瘤形成期间平滑肌细胞和巨噬细胞转化的作用。将在第3、7、14和28天测量主动脉直径。将收获主动脉组织以分析促炎细胞因子（IL-1β、TNF-α、MCP-1、IL-6和RANTES）、MMP-2和MMP-9活性（通过酶谱法）、丝氨酸蛋白酶（uPA、tPA和派-1）（通过蛋白质印迹法）、弹性蛋白和胶原降解以及主动脉平滑肌表达（通过组织学）和免疫细胞（巨噬细胞、T细胞、中性粒细胞、B细胞）浸润（通过流式细胞术）。结果：初步数据发现，在AAA形成中弹性蛋白酶灌注后14天，IL-1 R1 KO小鼠的动脉瘤形成显著减弱。在TAA形成前3天通过渗透泵给予阿那白滞素的野生型雄性小鼠也表现出主动脉直径减小。与野生型雄性小鼠相比，IL 1 β和IL 1 R1 KO雄性小鼠中的AAA形成显示主动脉直径减小、细胞因子产生减少（TNFα、MCP-1、IL-6和IL-27）、MMP-9表达减少以及巨噬细胞和中性粒细胞浸润减少。最后，通过qPCR和免疫组织化学，与非囊性对照相比，人AAA样本中的IL-1 R1增加。结论：IL-1途径抑制可以减轻局部弹性蛋白酶小鼠AAA模型中动脉瘤的形成和炎症。我们建议使用平滑肌细胞和巨噬细胞中的新谱系追踪分析来描绘AAA形成中的IL-1信号传导机制，并检查IL-1信号传导在动脉瘤形成的病理学中的作用，总体目标是开发动脉瘤治疗。