Myeloid KLF2 mediates S-nitrosylation-driven vascular response to ischemic injury

骨髓KLF2介导S-亚硝基化驱动的血管对缺血性损伤的反应

• 批准号: 9754870
• 负责人: David Ryan Sweet
• 金额: $ 5万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9754870
• 关键词: Address; Affect; Arteries; Attenuated; Biochemical; Biological Assay; Blood; Blood Vessels; Blood flow; Bypass; Caliber; Cardiovascular Diseases; Cell Lineage; Cell Proliferation; Cell Surface Receptors; Cell physiology; Cells; Cessation of life; Complex; Data; Disease; Distal; Endothelial Cells; Endothelium; Enzymes; Exposure to; Foundations; Functional disorder; Genetic; Genetic Transcription; Goals; Healthcare Systems; Hindlimb; Histologic; Image; Immune; Inflammation; Inflammatory; Injury; Intervention; Ischemia; Knock-out; Lead; Limb structure; Link; Liquid substance; Macrophage Activation; Maintenance; Mediating; Mediator of activation protein; Modeling; Modification; Molecular; Morbidity - disease rate; Mus; Myelogenous; Myeloid Cell Activation; Myeloid Cells; NOS2A gene; Nitric Oxide; Nitric Oxide Synthase; Nodal; Organ; Patients; Perfusion; Pharmacology; Phenotype; Physiological; Play; Process; Production; Proteins; Recovery; Regulation; Reperfusion Therapy; Role; Smooth Muscle Myocytes; Techniques; Tissues; Tumor-infiltrating immune cells; Vascular Smooth Muscle; Vascular remodeling; Vasodilation; Work; Zinc Fingers; adverse outcome; angiogenesis; capillary bed; imaging modality; improved; in vivo; in vivo Model; insight; macrophage; microCT; monocyte; mortality; prevent; programs; response; shear stress; tool; transcription factor

PROJECT SUMMARY/ABSTRACT Vascular occlusive disease poses an increasing burden on healthcare systems. Blockage of major arteries causes severe ischemic damage to tissues, with the potential of limb loss, organ dysfunction, and death. As a response to occlusion, however, collateral vessels are able to remodel to bypass the occlusion, a process termed arteriogenesis. Accumulating evidence implicates myeloid cells as key mediators of this remodeling, however the exact role these cells play in arteriogenesis remains unclear. Previous studies from our group have identified KLF2 as a transcriptional regulator of myeloid cell activation, a process critical for effective arteriogenesis. Nascent observations in our lab demonstrate that loss of KLF2 in the myeloid compartment greatly enhances perfusion recovery following a model of hindlimb ischemia (HLI). In addition, these differences appear to be greatly dependent on induction of the prominent inflammatory enzyme, inducible nitric oxide synthase (iNOS). Modification of critical proteins in vascular cells by S-nitrosylation has been shown to greatly affect functions crucial to vascular remodeling. These findings provide the basis for our central hypothesis that KLF2 serves as a regulator of macrophage-mediated nitric oxide (NO) production that affects the S-nitrosoproteome of endothelial and smooth muscle cells during occlusive injury. The proposed study will vigorously interrogate the effects of myeloid-KLF2 on vascular remodeling. Specifically, we aim to examine whether KLF2 serves as a nodal regulator of myeloid function during arteriogenesis, in part through its regulation of NO production. To accomplish this goal, this study will utilize a wide range of biochemical, molecular, and pharmacological techniques, including, but not limited to: in vivo models of arteriogenesis, microCT imaging, pharmacological modulation of NO, and S-nitrosylation assays. Together, these studies will provide crucial insight on the cellular and molecular processes leading to proper arteriogenesis and will provide the foundation for interventions targeting vascular occlusion with the goal of reducing debilitating complications for patients.

项目摘要/摘要 血管闭塞疾病给医疗系统带来了越来越大的负担。大动脉阻塞 对组织造成严重的缺血性损害，有可能导致肢体丧失、器官功能障碍和死亡。作为一名 对闭塞的反应，然而，侧支血管能够重塑以绕过闭塞，这是一个过程 称为动脉生成。越来越多的证据表明，髓系细胞是这种重塑的关键媒介， 然而，这些细胞在动脉形成中扮演的确切角色仍不清楚。我们小组之前的研究 已经确定KLF2是髓系细胞激活的转录调节因子，这一过程对于有效 动脉形成。我们实验室的初步观察表明，KLF2在髓鞘间丢失 极大地增强后肢缺血(HLI)模型后的血流灌注恢复。此外，这些 差异似乎在很大程度上取决于显著的炎性酶--诱导性一氧化氮的诱导。 一氧化氮合酶(INOS)。S亚硝化修饰血管细胞中的关键蛋白质 极大地影响对血管重塑至关重要的功能。这些发现为我们的中央 假设KLF2是巨噬细胞介导的一氧化氮(NO)产生的调节器，影响 闭合性损伤时内皮细胞和平滑肌细胞的S亚硝基蛋白质组。拟议的研究将 积极探讨髓系KLF2对血管重塑的影响。具体地说，我们的目标是研究 KLF2是否在动脉形成过程中作为髓系功能的结节调节器，部分是通过其 对NO生产的监管。为了实现这一目标，这项研究将利用广泛的生化， 分子和药理学技术，包括但不限于：动脉形成的活体模型， MicroCT成像、NO的药理调节和S亚硝化试验。总而言之，这些研究将 提供对导致适当动脉形成的细胞和分子过程的重要见解，并将提供 以减少衰弱并发症为目标的血管闭塞干预的基础 对病人来说。