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（由适用提供）：伤口检测和愈合当前的连续步骤是休眠形态发生程序，以恢复受伤后的障碍功能和组织稳态。白细胞在数百微米远的几秒钟内检测到伤口，并在几分钟内迁移到伤口。空间传播有关组织中损伤发生何时和何时发生的信息的机制几乎没有研究且被理解。通过协调受伤的组织中不同细胞类型的行为（包括白细胞，内皮细胞和上皮细胞），这些机制控制着炎症反应的长度和时间尺度，并保证炎症事件的持续时间和放大器（例如血管抑制，白细胞，术语等）量表。使用斑马鱼尾鳍图测定法，我们最近发现上皮氧化物DUOX会产生过氧化氢（H2O2）的梯度，该氢从伤口缘从伤口缘延伸到组织。这种梯度是快速伤口募集白细胞所必需的。但是，仍然不清楚反应性化学物质（例如H2O2）如何表现出很少的分子目标选择性并且会损害细胞的反应性化学物质是一种特定的伤口信号。我们假设在组织中，对H2O2的作用范围和/或细胞选择性的精确空间和临时控制允许其充当特定信号。到了解H2O2如何介导伤口检测，因此我们建议研究H2O2在何时何地产生H2O2，它在组织中传播的距离和快速，以及在何时何地，何时以及通过哪些信号通路不同的细胞类型对其做出响应。斑马鱼尾鳍绕组测定法代表了一种出色的脊椎动物模型系统，用于成像伤口反应以及通过药物和反向遗传学的分子扰动。为了系统地解决伤口反应的暂时和空间动力学，我们将使用无处不在，内皮，上皮，上皮和白细胞特定表达H2O2的转基因斑马鱼，用于H2O2的特定表达，它的上游激活量（Ca2+）和下游效果nf nf nf nf nf nf。关于多远和快速H2O2的基本问题，一种新型的旁分泌信号，在组织中传播，以及这种氧化化学物质如何能够介导特定的细胞反应。此外，我们将询问H2O2模式的长度和时间尺度如何由Duox Activator Ca2+调节。最后，我们将使用微阵列/生物信息学从转录签名中推断出H2O2下游教练或行动的途径。从中央炎症性调节剂NF知识开始，我们将对这些途径的时空激活进行成像，并通过H2O2梯度探测它们的调节。