Extracellular Histones in Burn-induced Microvascular Hyperpermeability

烧伤引起的微血管通透性过高中的细胞外组蛋白

基本信息

批准号：
10609034
负责人：
MACK H WU
金额：
$ 31.17万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10609034
关键词：
Acute Affect American Animals Anti-Inflammatory Agents Antibiotics Attenuated Binding Biological Assay Blood Blood Vessels Blood specimen Burn injury C Type Lectin Receptors Cannulations Cell Death Cell Nucleus Cell model Cells Cellular biology Cessation of life Characteristics Chromatin Circulation Confocal Microscopy Cultured Cells Cutaneous Data Development Edema Endothelium Environment Extracellular Space Extravasation Failure Fluid Therapy Focal Adhesions Functional disorder Goals Histamine Histones Hour Human Hypertension Imaging Techniques Immune Targeting Immunity Incineration Individual Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injury Intercellular Junctions Intra-abdominal Knowledge Life Lipids Lung Mediating Mediator Mesentery Microcirculation Molecular Molecular Conformation Mus Nuclear Nuclear Proteins Organ PTK2 gene Pathologic Pathology Pathway interactions Patients Permeability Pharmaceutical Preparations Phosphorylation Plasma Production Protein Tyrosine Kinase Receptor Signaling Reporting Research Role Series Signal Pathway Signal Transduction Study Subject Syndrome Techniques Testing Tissues Trauma Vascular Endothelial Cell Vascular Endothelium Visceral burn therapy cell injury comparative cytokine design experimental study extracellular healthy volunteer human subject in vivo in vivo evaluation inhibitor injured innovation insight monolayer mortality mouse model nanoimaging neutralizing antibody novel novel diagnostics novel therapeutics optoacoustic tomography organ injury receptor respiratory distress syndrome response severe burns systemic inflammatory response theories tissue injury translational approach translational study treatment strategy ultra high resolution vascular injury

项目摘要

ABSTRACT Extracellular histones are nuclear proteins released to the extracellular environment during tissue destruction or injury. Emerging evidence implicates them as danger associated molecules with immunostimulatory capability. The receptor-signaling mechanisms responsible for their tissue or cell-specific effects are poorly understood. In recent studies, we detected elevated plasma levels of histones in burn patients as well as animals. Administration of histones caused microvascular leakage and endothelial hyperpermeability-characteristic pathology underlying multiple organ dysfunction following burns, whereas histone inhibitors attenuated burn-induced barrier leakage. Moreover, we obtained novel evidence for the critical role of C-type lectin receptor 2d (Clec2d)-mediated tyrosine kinase signaling in endothelial response to histones. Built on these intriguing findings, this study will characterize the release, pathophysiological function, and molecular mechanisms of histones as an important contributor to burn-induced endothelial barrier injury in edema-prone tissues, including lungs and gut/mesenteric microvessels. We propose three aims: Aim 1 to characterize circulating histones in burn patients and animals correlated with organ dysfunction; Aim 2 to determine the causal effects of histones on microvascular hyperpermeability during burns; Aim 3 to explore the molecular mechanisms by which histones induce endothelial barrier breakdown. The specific mechanistic hypothesis to be tested is that following thermal destruction of tissues, injured cells release histones into the circulation where they directly interact with the vascular endothelium by binding to Clec2d and activating downstream intracellular signaling mediated by Syk/Src-FAK; these tyrosine kinases phosphorylate proteins that constitute cell-cell junctions and cell-matrix focal adhesions, thereby triggering their conformational changes and leading to increased permeability. This novel pathway will be tested in innovative experiments that incorporate newly developed imaging techniques and molecular assays into a comparative analysis of burn patients, human organs, and animal/cell models. Through this translational study, we expect to gain new insights that will not only shift the current paradigms in vascular endothelial cell biology, but also fill the gaps of knowledge in understanding burn pathophysiology. Identification of circulating histones as a key mediator of burn-induced tissue/organ injury may lead to the development of new diagnostics and therapies for thermal trauma.

抽象的细胞外组蛋白是在组织破坏或组织破坏过程中释放到细胞外环境的核蛋白。受伤。新的证据表明它们是具有免疫刺激能力的危险相关分子。对其组织或细胞特异性作用的受体信号传导机制知之甚少。在最近的研究中，我们检测到烧伤患者和动物的血浆组蛋白水平升高。行政组蛋白引起微血管渗漏和内皮细胞通透性过高-潜在的特征性病理学烧伤后多器官功能障碍，而组蛋白抑制剂则减弱烧伤引起的屏障渗漏。此外，我们获得了 C 型凝集素受体 2d (Clec2d) 介导的酪氨酸关键作用的新证据内皮细胞对组蛋白反应中的激酶信号传导。基于这些有趣的发现，本研究将描述组蛋白的释放、病理生理功能和分子机制作为重要贡献者烧伤引起的易水肿组织的内皮屏障损伤，包括肺和肠/肠系膜微血管。我们提出三个目标：目标 1 表征烧伤患者和与烧伤相关的动物的循环组蛋白器官功能障碍；目标 2 确定组蛋白对微血管通透性过高的因果影响烧伤；目标 3 探索组蛋白诱导内皮屏障破坏的分子机制。这待测试的具体机制假设是，在组织热破坏后，受损细胞释放组蛋白进入循环系统，通过与 Clec2d 结合直接与血管内皮细胞相互作用激活 Syk/Src-FAK 介导的下游细胞内信号传导；这些酪氨酸激酶磷酸化构成细胞-细胞连接和细胞-基质粘着斑的蛋白质，从而触发其构象变化并导致渗透性增加。这一新途径将在创新实验中进行测试将新开发的成像技术和分子测定纳入烧伤的比较分析中患者、人体器官和动物/细胞模型。通过这项转化研究，我们期望获得新的见解这不仅将改变血管内皮细胞生物学的当前范式，而且还将填补知识空白了解烧伤病理生理学。鉴定循环组蛋白作为烧伤诱导的关键介质组织/器官损伤可能会导致热损伤新诊断和治疗方法的发展。