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期间释放到细胞外环境的核蛋白质或受伤。新兴的证据暗示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负责的受体信号机制知之甚少。在最近的研究，我们检测到燃烧患者和动物的血浆组蛋白的血浆水平升高。行政组蛋白引起微血管泄漏和内皮过度透明度 - 特征病理燃烧后多个器官功能障碍，而组蛋白抑制剂减弱了烧伤引起的屏障泄漏。此外，我们获得了C型凝集素受体2D（CLEC2D）介导的酪氨酸的关键作用的新证据内皮反应中的激酶信号传导。这项研究以这些有趣的发现为基础组蛋白的释放，病理生理功能和分子机制是重要的促进者燃烧诱导的容易浮肿组织的内皮屏障损伤，包括肺和肠/肠系膜微血管。我们提出了三个目标：目标1来表征与烧伤患者和动物相关的烧伤组蛋白的表征器官功能障碍；目标2以确定组蛋白对微血管高温性的因果影响烧伤；目标3探索组蛋白诱导内皮屏障分解的分子机制。这要测试的具体机械假设是，在组织热破坏后，受伤的细胞释放组蛋白进入循环系统，通过与Clec2d结合，它们直接与血管内皮相互作用激活由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 - 细胞连接和细胞矩阵局灶性粘附的蛋白质，从而触发其构象变化并导致渗透率增加。这种新颖的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燃烧的比较分析患者，人体器官和动物/细胞模型。通过这项翻译研究，我们希望获得新的见解这不仅会改变血管内皮细胞生物学中当前的范例，而且还会填补知识的空白了解烧伤病理生理学。鉴定循环组蛋白是燃烧引起的关键介体组织/器官损伤可能会导致热创伤的新诊断和疗法的发展。