Nitric Oxide Signaling in Hypoxia and Immunity in Drosophila

果蝇缺氧和免疫中的一氧化氮信号传导

基本信息

批准号：
7694365
负责人：
PATRICK H O'FARRELL
金额：
$ 34.13万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-03-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7694365
关键词：
Acute Address Animal Model Area Biodiversity Biological Biological Models Cell Culture Techniques Cell Survival Cells Coupling Cultured Cells Cysteine Cytoplasm Defect Development Dissection Drosophila genus Embryo Event Experimental Models Generations Genes Genetic Genetic Epistasis Genetic Models Guanylate Cyclase Health Human Hypoxia Hypoxia Inducible Factor Immune Immune response Immunity Infection Interphase Cell Intracellular Transport Investigation Ischemia Larva Life Measures Mediating Mediator of activation protein Mitochondria Modeling Mutation Myocardial Infarction Natural Immunity Nature Nitric Oxide Nitric Oxide Pathway Nitric Oxide Signaling Pathway Nitric Oxide Synthase Nitrites Organ Transplantation Oxygen Pathway interactions Permeability Personal Satisfaction Phenotype Physiological Play Post-Translational Protein Processing Process Processed Genes Production Proteins RNA Interference Reactive Oxygen Species Reporter Role Signal Pathway Signal Transduction Signal Transduction Pathway Source Stress Stroke Swelling System Testing Thioredoxin Tissues Transcriptional Regulation Transducers Transgenes animation base cancer therapy chemotherapy computerized data processing deprivation gene discovery hemodynamics in vivo man novel prevent public health relevance response tool tumor growth

项目摘要

DESCRIPTION (provided by applicant): Hypoxia figures importantly in the biggest of health issues. It is responsible for the damage caused by the ischemia accompanying cardiac infarct and stroke, and it plays a central role in limiting tumor growth as well as blunting the actions of important chemotherapies. While we have a mechanistic understanding of hypoxic regulation of transcription by Hypoxia Inducible Factor (HIF), other modes of response to oxygen deprivation are still poorly understood. How does the mitochondrion, the major consumer of oxygen in the cell, adjust to shortfalls in oxygen supply, and how does it signal to the rest of the cell, demanding accommodations to these shortfalls? Reactive oxygen species produced by mitochondria have been ascribed the responsibility of communicating oxygen stress to the cell, but precise pathways of signal generation, transduction and action have not been established. Recognizing the complexity of the issues and the fundamental nature of the questions, it seems that the powerful genetics available in model organisms could make a major contribution. We found that nitric oxide (NO) mediates immediate responses to hypoxia in Drosophila, including an arrest of embryos in a reversible state of suspended animation. We have devoted ourselves to the development of a powerful and greatly simplified experimental context in which we can dissect the NO-signaling mechanisms. Our finding that NO regulates innate immune responses in Drosophila revealed a hypoxia-immunity connection, and gave us the simplification we sought. Immune reporters signal the action of NO, and the response to hypoxia can be recapitulated in cultured cells amenable to powerful RNAi screens. In this proposal, we will combine in vivo studies and analysis of cell culture models to investigate NO-mediated signaling. In vivo studies will probe how infection induces NOS, and how NO signaling contributes to the immune response. In Drosophila S2 cells, we will examine the basis of hypoxia-induced NO signaling that appears to originate in the mitochondria, and will trace the transformation and transport of this signal as it is conveyed to the cytoplasm. Finally, we will define the genes and pathways by which mitochondria sense hypoxia and signal the dramatic changes within the mitochondria, and will the coupling between mitochondrial changes and the signals emanating from the mitochondria that have such huge impacts on survival of cells and the well-being of man. These studies will give us a mechanistic understanding of a major component of the hypoxia response, contribute to the understanding of the biological diversity in tolerance to hypoxia, and give us approaches to manipulate hypoxia sensitivity to potentially benefit issues of important health concern. PUBLIC HEALTH RELEVANCE The most common life-threatening health problems, cardiac infarct and stroke, cause damage by interrupting oxygen supply, yet we understand little about the biological responses to the resulting acute hypoxia. We will dissect the mechanisms of response to hypoxia in a powerful model genetic system, Drosophila. Our findings have the potential to influence treatment of cardiac infarct and stroke, and could have an impact on areas as diverse as sustaining organs for transplantation, and cancer therapies.

描述（由申请人提供）：在最大的健康问题中重要的缺氧数字。它是由伴随心脏梗塞和中风的缺血造成的损害，并且在限制肿瘤生长以及削弱重要化学疗法的作用方面起着核心作用。虽然我们对缺氧诱导因子（HIF）对转录的低氧调节有一种机械理解，但其他对氧气剥夺的反应方式仍然很少了解。线粒体是细胞中的主要氧气消费者，适应氧气供应的不足，以及如何向其他细胞发出信号，要求对这些不足的适应性？线粒体产生的活性氧已被归因于将氧应激传达给细胞的责任，但尚未确定信号产生，转导和作用的精确途径。认识到问题的复杂性和问题的基本性质，似乎模型生物体中可用的强大遗传学可以做出重大贡献。我们发现，一氧化氮（NO）介导了果蝇中缺氧的立即反应，包括在可逆的悬浮动画状态下逮捕胚胎。我们致力于发展强大而简化的实验环境，在这种情况下，我们可以剖析无信号的机制。我们的发现没有调节果蝇中先天免疫反应显示出缺氧 - 免疫的联系，并为我们提供了简化的信息。免疫记者向NO的作用发出了信号，并且可以在培养的细胞中概括对强大的RNAi筛查的培养细胞的反应。在此提案中，我们将结合体内研究和细胞培养模型的分析，以研究无介导的信号传导。体内研究将探测感染如何诱导NOS，以及无信号传导如何促进免疫反应。在果蝇S2细胞中，我们将检查缺氧诱导的无信号传导的基础，该信号似乎起源于线粒体，并将追踪该信号传递到细胞质时的转化和运输。最后，我们将定义线粒体感知缺氧的基因和途径，并向线粒体内部的急剧变化发出巨大变化，并将在线粒体变化和线粒体发出的信号之间的耦合对细胞的存活以及良好的生存产生巨大影响是人。这些研究将使我们对缺氧反应的主要组成部分有一种机械理解，有助于理解对低氧耐受性的生物学多样性，并为我们赋予操纵缺氧对潜在健康问题的潜在问题的敏感性的方法。公共卫生相关性是最常见的威胁生命的健康问题，心脏梗死和中风，通过中断氧气供应而造成损害，但我们对生物学对急性缺氧的生物学反应一无所知。我们将在强大的模型遗传系统果蝇中剖析对缺氧反应的机制。我们的发现有可能影响心脏梗塞和中风的治疗，并且可能会影响如维持移植器官和癌症疗法等多样性的区域。