Zebrafish as a natural host model for Vibrio cholerae

斑马鱼作为霍乱弧菌的天然宿主模型

• 批准号: 8160606
• 负责人: JEFFREY H WITHEY
• 金额: $ 22.8万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-07 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8160606
• 关键词: Animal Model; Animals; Area; Bacteria; Biological Models; Biological Process; Cessation of life; Cholera; Cholera Toxin; Complex; Conditioned Culture Media; Data; Developing Countries; Diarrhea; Disease; Disease Outbreaks; Disease model; Dose; Environment; Epidemic; Factor V; Fishes; Future; Gastrointestinal tract structure; Genes; Genetic Transcription; Goals; Human; Immune response; Incubated; Infant; Infection; Insecta; Intestines; Learning; Life; Life Style; Mediating; Modeling; Mus; Natural Disasters; Organism; Oryctolagus cuniculus; Pathogenesis; Pathway interactions; Patients; Pilum; Plankton; Process; Public Health; Regulon; Research; Research Personnel; Role; Shellfish; Signal Transduction; Site; Study models; Swimming; System; Testing; Time; TimeLine; Toxin; Vibrio cholerae; Vibrio cholerae O1; Vibrio cholerae non-O1; Virulence; Virulence Factors; War; Water; Work; Zebrafish; design; egg; in vivo; infectious disease model; killings; mouse model; neonate; new therapeutic target; pandemic disease; pathogen; research study

DESCRIPTION (provided by applicant): Vibrio cholerae are aquatic bacteria that cause the severe diarrheal disease cholera when ingested by humans in contaminated water. Cholera afflicts an estimated 5 million people annually, causing over 100,000 deaths. Currently the predominant animal models for V. cholerae are infant mice and rabbits. However, these models are expensive, difficult, unpleasant, and do not reproduce true disease states, as they are not natural hosts for V. cholerae. V. cholerae naturally inhabit several aquatic niches, including associations with shellfish, insect egg masses, and plankton. Recently V. cholerae were also found to colonize the intestinal tracts of vertebrate fish. Our central hypothesis is that the zebrafish, Danio rerio, is an ideal natural animal host model for V. cholerae that will make new studies of both pathogenesis and environmental colonization possible. Zebrafish have been utilized extensively in the past 20 years as model organisms for many different biological processes, including infectious disease models. Preliminary data indicate that V. cholerae colonize the zebrafish intestine to very high levels when administered either directly into the digestive tract via gavage or when simply added to water in which zebrafish are incubated. The latter is a much more natural, representative model for colonization than any existing animal model, and we are likely to learn important new information about the colonization process by utilizing zebrafish as an animal model. In addition to colonizing the zebrafish intestine, V. cholerae that have a functional virulence regulatory system also kill zebrafish through an unknown mechanism. Therefore, zebrafish may also be a good model for studying V. cholerae disease processes. The objective of this proposal is to establish zebrafish as a functional natural animal model for V. cholerae. We will determine the sites of V. cholerae colonization of zebrafish. We will establish a timeline for colonization, such that future experiments can utilize the most relevant time points for collecting data. We will determine the infectious dose required for V. cholerae colonization of zebrafish, which is important for the design of future experiments and will illuminate the dynamics of free swimming V. cholerae and fish in the natural environment. Finally, we will determine how secreted factors from V. cholerae cause fish death. Because the major virulence regulon is required for causing zebrafish death, V. cholerae virulence genes must be induced in the presence of zebrafish. Our long term goal is to identify the signals that induce virulence during human infection such that virulence can be disrupted therapeutically. The zebrafish model presents an entirely new opportunity for identifying these signals. Establishment of zebrafish as a natural animal host model should prove extremely useful to V. cholerae researchers studying not only colonization and disease processes but also the environmental lifestyle of V. cholerae, which is very important for understanding the V. cholerae reservoirs that can produce epidemic or pandemic cholera outbreaks. Therefore establishment of this new model should have a broad impact on V. cholerae research worldwide. PUBLIC HEALTH RELEVANCE: Cholera is a major public health problem in many developing countries and areas suffering from the effects of war or natural disaster, with an estimated 5,000,000 new cases and 100,000 deaths each year. The goal of this proposal is to establish zebrafish as a new natural host model for V. cholerae, the cause of cholera. This new model will facilitate better understanding of both how V. cholerae live in the environment and how they become human pathogens.

描述（由申请人提供）：霍乱弧菌是一种水生细菌，当人类在受污染的水中摄入时，会引起严重的霍乱。据估计，霍乱每年影响500万人，造成10万多人死亡。目前，霍乱弧菌的主要动物模型是幼年小鼠和兔。然而，这些模型是昂贵的，困难的，令人不快的，并且不能再现真实的疾病状态，因为它们不是霍乱弧菌的天然宿主。霍乱弧菌自然栖息于几个水生生态位，包括与贝类，昆虫卵块和浮游生物的联系。最近还发现霍乱弧菌在脊椎动物的肠道中定植。我们的中心假设是，斑马鱼，斑马鱼，是一个理想的自然动物宿主模型的霍乱弧菌，将使新的研究的发病机制和环境殖民可能。在过去的20年里，斑马鱼被广泛用作许多不同生物过程的模式生物，包括传染病模型。初步数据表明，当通过管饲法直接施用到消化道中或当简单地添加到孵育斑马鱼的水中时，霍乱弧菌在斑马鱼肠道中定殖至非常高的水平。后者是一个更自然的，代表性的殖民模型比任何现有的动物模型，我们可能会学习重要的新信息，通过利用斑马鱼作为动物模型的殖民过程。除了在斑马鱼肠道中定居外，具有功能性毒力调节系统的霍乱弧菌还通过未知机制杀死斑马鱼。因此，斑马鱼也可能是研究霍乱弧菌疾病过程的良好模型。本研究的目的是建立斑马鱼作为霍乱弧菌的功能性天然动物模型。我们将确定斑马鱼的霍乱弧菌定殖位点。我们将建立定植的时间轴，以便将来的实验可以利用最相关的时间点收集数据。我们将确定斑马鱼的霍乱弧菌定殖所需的感染剂量，这对未来实验的设计很重要，并将阐明自然环境中自由游泳的霍乱弧菌和鱼类的动态。最后，我们将确定霍乱弧菌的分泌因子如何导致鱼类死亡。由于主要毒力调节子是导致斑马鱼死亡所必需的，因此必须在斑马鱼存在的情况下诱导霍乱弧菌毒力基因。我们的长期目标是鉴定在人类感染期间诱导毒力的信号，使得毒力可以在治疗上被破坏。斑马鱼模型为识别这些信号提供了一个全新的机会。斑马鱼作为一种自然动物宿主模型的建立将被证明对霍乱弧菌研究人员非常有用，不仅研究霍乱弧菌的定殖和疾病过程，而且研究霍乱弧菌的环境生活方式，这对于了解可能导致流行性或大流行性霍乱暴发的霍乱弧菌宿主非常重要。因此，这一新模型的建立将对世界范围内霍乱弧菌的研究产生广泛影响。 公共卫生相关性：霍乱是许多发展中国家和受战争或自然灾害影响的地区的一个主要公共卫生问题，估计每年有500万新病例和10万人死亡。该提案的目标是将斑马鱼建立为霍乱弧菌（霍乱的原因）的新天然宿主模型。这一新模型将有助于更好地了解霍乱弧菌如何在环境中生存以及它们如何成为人类病原体。