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ROS responses during Vibrio cholerae infection

霍乱弧菌感染期间的ROS反应

基本信息

批准号：
9102467
负责人：
Jun Zhu
金额：
$ 50.33万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9102467
关键词：
Adult Animal Model Animals Cell physiology Cells Cholera Country Cysteine Disease Enzymes Escherichia coli Experimental Animal Model Feces Fimbriae Proteins Gene Expression Genes Goals Growth Haiti Human Immune Immune response Infection Inflammation Inflammatory Intestines Lead Light Minor Modeling Modification Mus NADPH Oxidase Pathogenesis Pathogenicity Pathology Patients Phase Play Post-Translational Protein Processing Post-Translational Regulation Poverty Production Property Proteins Proteomics Reactive Oxygen Species Regulation Regulatory Pathway Research Resistance Resolution Resources Role Salmonella Sampling Series Signal Pathway Signal Transduction Small Intestines Sulfhydryl Compounds Tissues Transcriptional Regulation Vibrio cholerae Virulence Virulence Factors Work base coping gut microbiome gut microbiota in vivo microbial microbiome mouse model novel novel strategies pathogen public health relevance research study resistance mechanism response tissue culture transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Vibrio cholerae is a human pathogen which colonizes small intestines of host, resulting in the onset of a severe diarrheal disease known as cholera. In order for V. cholerae to successfully colonize the host, it must express a series of virulence factors, which have been the main focus of the cholera research. However, bacterial pathogenicity is a multifactorial property in vivo that involves host response to infection, and gu microbiome interference of colonization. For example, although the pathology of cholera is not immune driven, it has been shown that minor, but significant inflammation responses in cholera patients and in experimental animal models are induced by V. cholerae infection and likely play a role in the resolution of disease. Little is known how inflammation is induced by V. cholerae and how V. cholerae copes with these signals and help its colonization. Here we performed an RNAseq analysis on mouse intestines free of V. cholerae and those that were infected to determine host responses to V. cholerae infection. One gene involved in reactive oxygen species (ROS) production, Duox2, encoding an NADPH oxidase and has been shown to be essential for controlling the growth of gut flora, was strongly upregulated. We show that both Duox2 expression and ROS production are induced by V. cholerae in a tissue culture model and in mice. Interestingly, this induction is dependent on V. cholerae virulence gene expression. Moreover, our preliminary studies indicate that V. cholerae cells in cholera patients' stool samples are highly resistant to ROS, suggesting that V. cholerae undergoes induction of ROS resistance during infection. To elucidate how V. cholerae overcome ROS produced by the host, we performed a Tn-seq experiment and found a set of V. cholerae genes that are required for ROS resistance in vivo. Together with a proteomics approach, we reveal novel transcriptional and posttranslational regulation mechanisms that are involved in regulating ROS resistance. Therefore we hypothesize that during infection, V. cholerae induces host Duox2 expression, thus ROS production, which facilitate V. cholerae colonization by modulating gut flora composition, whereas transcriptional and posttranslational regulatory pathways lead to V. cholerae inherently resistant to ROS in vivo. We will investigate the mechanism of V. cholerae-induced host ROS production and its effects on gut microbiota composition and V. cholerae colonization. We will investigate V. cholerae ROS resistance during infection of an adult mouse model. We will focus on transcriptional regulation and posttranslational modification of cellular functions of ROS resistance. Finally, we will investigate V. cholerae ROS resistance mechanisms in cholera patients by performing RNAseq and proteomic analysis of V. cholerae cells directly from cholera patient vomituses and stools.

描述(申请人提供)：霍乱弧菌是一种人类病原体，它定植于宿主的小肠，导致一种被称为霍乱的严重腹泻疾病的发病。在……里面霍乱弧菌要想在宿主上成功定植，必须表达一系列毒力因子，这些毒力因子一直是霍乱研究的重点。然而，细菌的致病力在体内是一个多因素的特性，涉及宿主对感染的反应，以及GU微生物群对定植的干扰。例如，虽然霍乱的病理不是由免疫驱动的，但已表明霍乱患者和实验动物模型中轻微但显著的炎症反应是由霍乱弧菌感染引起的，并可能在疾病的解决中发挥作用。霍乱弧菌是如何引起炎症的，以及霍乱弧菌如何应对这些信号并帮助其定植，目前还知之甚少。在这里，我们对没有霍乱弧菌的小鼠肠道和感染霍乱弧菌的小鼠肠道进行了RNAseq分析，以确定宿主对霍乱弧菌感染的反应。参与ROS产生的一个基因DUOX2编码NADPH氧化酶，已被证明是控制肠道菌群生长所必需的，它被强烈上调。我们发现霍乱弧菌在组织培养模型和小鼠体内都能诱导DUOX2的表达和ROS的产生。有趣的是，这种诱导依赖于霍乱弧菌毒力基因的表达。此外，我们的初步研究表明，霍乱患者粪便中的霍乱弧菌细胞对ROS具有高度的抗性，这表明霍乱弧菌在感染过程中经历了对ROS的诱导耐药。为了阐明霍乱弧菌如何克服宿主产生的ROS，我们进行了TN-seq实验，并在体内找到了一组抵抗ROS所需的霍乱弧菌基因。结合蛋白质组学方法，我们揭示了参与调节ROS抗性的新的转录和翻译后调节机制。因此我们推测，在感染过程中，霍乱弧菌诱导宿主DUOX2的表达，从而产生ROS，从而通过调节肠道菌群组成促进霍乱弧菌的定植，而转录和翻译后调控途径导致霍乱弧菌在体内对ROS产生固有的抗性。我们将研究霍乱弧菌诱导宿主ROS产生的机制及其对肠道微生物区系组成和霍乱弧菌定植的影响。我们将在成年小鼠模型上研究霍乱弧菌对ROS的耐药性。我们将重点介绍ROS抗性细胞功能的转录调控和翻译后修饰。最后，我们将通过对霍乱患者呕吐物和粪便中的霍乱弧菌细胞进行RNAseq和蛋白质组学分析来研究霍乱患者对ROS的耐药性机制。