Regulation of Wound Detection in Animal Tissues

动物组织伤口检测的监管

基本信息

批准号：
8545186
负责人：
Philipp Michael Niethammer
金额：
$ 33.53万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-15 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8545186
关键词：
Address Affect Back Behavior Binding Sites Bioinformatics Biological Assay Biological Models Biosensor Blood Vessels Calcium Cell Nucleus Cells Chemicals Chemotactic Factors Cluster Analysis Data Dependency Detection Diffuse Disease Endothelial Cells Epithelial Epithelial Cells Event Exhibits Fishes Fluorescence Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Transcription Healed Homeostasis Human Hydrogen Peroxide Image Inflammatory Inflammatory Response Injury Lead Length Leukocytes Life Literature Malignant Neoplasms Mediating Molecular Molecular Target NADPH Oxidase Nucleic Acid Regulatory Sequences Orthologous Gene Paracrine Communication Pathologic Pathway interactions Pattern Pharmacology Physiological Production Recruitment Activity Regulation Reporter Serum Response Factor Signal Pathway Signal Transduction Site Stream Tail Testing Time Tissues Transgenic Organisms Travel Vasodilation Zebrafish animal tissue autocrine cell injury cell type extracellular healing in vivo migration molecular imaging novel paracrine positional cloning programs response spatiotemporal wound

项目摘要

DESCRIPTION (provided by applicant): Wound detection and healing present consecutive steps of a dormant morphogenetic program to restore barrier function and tissue homeostasis after injury. Leukocytes detect a wound within seconds from hundreds of micrometers away, and migrate to the wound within minutes. The mechanisms that spatially propagate the information on where and when an injury has occurred in a tissue remain little studied and understood. By coordinating the behavior of different cell types in the wounded tissue (incl. leukocytes, endothelial and epithelial cells), these mechanisms control length- and time- scales of inflammatory responses, and warrant that duration and amplitude of inflammatory events (e.g. vasodilation, leukocyte recruitment, etc.) scale appropriately with the extent of tissue damage. Using the zebrafish tail fin wounding assay, we recently found that the epithelial NADPH oxidase DUOX generates a gradient of hydrogen peroxide (H2O2) that extends up to ~200 um from the wound margin into the tissue. This gradient is required for rapid wound recruitment of leukocytes. However, it remains still unclear how a reactive chemical such as H2O2, which exhibits little molecular target selectivity and that can damage cells, is harnessed as a specific wound signal. We hypothesize that within tissues, the precise spatial and temporal control of H2O2's range of action and/or cell selectivity allows it to act as a specific signal. To understand how H2O2 mediates wound detection, we thus propose to investigate where and when H2O2 is generated, how far and fast it propagates through the tissue, and where, when, and via which signaling pathways different cell types respond to it. The zebrafish tail fin wounding assay represents an excellent vertebrate model system for imaging wound responses and for molecular perturbation by pharmacology and reverse genetics. To systematically address temporal and spatial dynamics of wound responses, we will use transgenic zebrafish with ubiquitous, endothelial, epithelial, and leukocyte specific expression of fluorescence reporters for H2O2, its likely upstream activator calcium (Ca2+), and downstream effectors NF¿B. Using biosensor imaging and molecular perturbation in live zebrafish, we will address fundamental questions of how far and fast H2O2, a novel paracrine signal, travels in tissues, and how this oxidizing chemical is able to mediate specific cellular responses. Further, we will interrogate how length and timescales of H2O2 patterns are regulated by the DUOX activator Ca2+. Finally, we will deduce pathways that cooperate or act downstream of H2O2 from their transcriptional signature using microarray/bioinformatics. Starting with NF¿B, a central inflammatory regulator, we will image the spatiotemporal activation of these pathways, and probe their regulation by the H2O2 gradient.

描述（由申请人提供）：伤口检测和愈合是休眠形态发生程序的连续步骤，用于在损伤后恢复屏障功能和组织稳态。白细胞在几秒钟内检测到数百微米外的伤口，并在几分钟内迁移到伤口。通过协调受伤组织中不同细胞类型（包括白细胞，内皮细胞和上皮细胞），这些机制控制炎症反应的长度和时间尺度，并保证炎症事件（例如血管舒张、白细胞募集等）的持续时间和幅度与组织尾鳍损伤的程度适当地缩放。分析中，我们最近发现上皮 NADPH 氧化酶 DUOX 产生的过氧化氢 (H2O2) 梯度可达约 200从伤口边缘进入组织需要这种梯度。然而，目前尚不清楚如何利用诸如 H2O2 之类的反应性化学物质作为一种特定的分子靶点选择性。我们追求的是，在组织内，H2O2 的作用范围和/或细胞选择性的精确空间和时间控制使其能够充当特定信号。了解 H2O2 如何介导伤口检测，因此我们建议研究 H2O2 产生的地点和时间，它在组织中传播的距离和速度，以及不同细胞类型在何处、何时以及通过哪些信号通路对其做出反应。伤口测定代表了一种优秀的脊椎动物模型系统，用于成像伤口反应以及通过药理学和反向遗传学进行分子扰动为了系统地解决伤口反应的时间和空间动态，我们将使用转基因斑马鱼。 H2O2 荧光产生剂普遍存在，内皮、上皮和白细胞特异性表达，其可能的上游激活剂钙 (Ca2+) 和下游效应器 NF¿ B. 利用活体斑马鱼的生物传感器成像和分子扰动，我们将解决 H2O2（一种新型旁分泌信号）在组织中传播的距离和速度的基本问题，以及这种氧化化学物质如何能够介导特定的细胞反应。探究 DUOX 激活剂 Ca2+ 如何调节 H2O2 模式的长度和时间尺度。最后，我们将从 H2O2 的下游合作或作用途径中推断出它们。使用微阵列/生物信息学的转录签名从 NF¿ B 是一种中央炎症调节因子，我们将对这些通路的时空激活进行成像，并通过 H2O2 梯度探究它们的调节。