Detection of Obesogens and Diabetogens by Zebrafish Screening Models

描述（由申请人提供）：肥胖和肥胖相关疾病，如2型糖尿病、高血压和心血管疾病，在过去几十年中在西方国家，特别是美国急剧增加。这种增加的原因可能是多因素的;其中一个建议的新原因是加速暴露于肥胖或糖尿病诱导化学品（致肥胖剂或致糖尿病剂）。然而，在人类接触的数万种工业化学品中，很少有对致肥胖或致糖尿病能力进行过测试，主要是由于适当的筛选模型有限。在这里，我们建议开发基于斑马鱼的屏幕，以确定obesogens和diabetogens和确定剂量反应效应和曝光的关键窗口。本研究的目的是：1）建立基于脂质结合荧光染料的斑马鱼肥胖基因筛选方法; 2）建立基于转基因斑马鱼的斑马鱼糖尿病基因筛选方法; 3）研究肥胖基因和糖尿病基因在斑马鱼体内的作用机制。该项目的预期成果是用于肥胖和糖尿病筛查的中高通量模型，利用外部（子宫外），快速和透明的胚胎发育以及斑马鱼筛查的成本效益。这符合国家毒理学计划（NTP）的愿景，如在其21世纪世纪路线图中所述，以实现NTP愿景“完善传统毒理学测定，开发环境诱导疾病的快速、基于机制的预测筛选”（US National Toxicology Roadmap“A national toxicology program for the 21st世纪”;：ntp.niehs.nih.gov/files/ NTPrdmp.pdf）。我们将确定致肥胖和致糖尿病，暴露的临界窗口和效应剂量。通过将我们生成的结果和模型纳入NTP和EPA在美国开展的其他化学品筛选活动，并通过与化学品安全领域的其他举措合作，我们将参与大型筛选 努力确定化学品的优先次序，以便进一步进行风险评估。该项目的最终影响是保护人口在生命的关键时期免受致肥胖和致糖尿病物质的暴露，这符合NIH的总体使命，即通过预防疾病保护人类健康，保护发育中的胎儿，并增加对环境污染物生物效应的了解（NIH使命;：//www.nih.gov/about/mission.htm）。此外，该提议探索了斑马鱼模型作为毒性筛选的工具，这得到了NIH对斑马鱼和其他动物模型系统用于生物医学和行为研究的强烈倡导的支持（Henken DB，Rasooly RS，Javois L，休伊特AT. 2004年，美国国立卫生研究院跨NIH斑马鱼协调委员会。美国国立卫生研究院和斑马鱼的生长作为一个实验模式生物。斑马鱼1（2）：105-10）。! 公共卫生相关性：肥胖和肥胖相关疾病，如2型糖尿病，高血压和心血管疾病，在西方国家，特别是美国在过去几十年中急剧增加。一个新出现的原因可能是我们暴露在促进肥胖的工业化学品中。然而，在我们接触到的成千上万种化学物质中，很少有人测试过它们导致肥胖或糖尿病的能力，这主要是由于缺乏筛选这些化合物的模型。在这个项目中，我们将建立一个筛选模型，以识别导致肥胖和糖尿病的化学物质。筛选模型是基于斑马鱼，尽管它们与人类有明显的差异，但它们具有相似的制造和储存脂肪以及产生胰岛素的生物学方式。这些模型将用于筛选化学品和预测人类接触化学品的风险。!

DESCRIPTION (provided by applicant): Obesity and obesity-related disorders, such as type 2 diabetes, hypertension, and cardiovascular disease, have increased dramatically in Western countries, particularly the United States during the past decades. The reasons for this increase are likely to be multifactorial; one of the suggested emerging causes being the accelerated exposure to obesity or diabetes-inducing chemicals (obesogens or diabetogens). However, out of the tens of thousands of industrial chemicals that humans are exposed to, very few have been tested for obesogenic or diabetogenic capacity, mostly due to the limited availability of appropriate screening models. We here propose to develop zebrafish-based screens to identify obesogens and diabetogens and determine dose-response effects and critical windows of exposure. The aims are 1) To develop the zebrafish obesogen screen based on lipid binding fluorochromes 2) To establish the zebrafish diabetogen screen based on transgenic zebrafish, and 3) To characterize the mechanisms of action of obesogens and diabetogens in zebrafish. The expected outcomes of this project are medium to high throughput models for obesogen and diabetogen screening, taking advantage of the external (ex-utero), rapid and transparent embryonic development as well as cost efficiency of zebrafish screens. This is in line with the vision of the National Toxicology Program (NTP) as described in its 21st Century Roadmap to Achieve the NTP Vision in "refining traditional toxicology assays, developing rapid, mechanism-based predictive screens for environmentally induced diseases" (US National Toxicology Roadmap "A national toxicology program for the 21st century"; ://ntp.niehs.nih.gov/files/ NTPrdmp.pdf). We will identify obesogens and diabetogens, critical windows of exposure and effect doses. By incorporating of our generated results and models into other chemical screening activities in the US, performed by NTP and EPA, and through collaborations with other initiatives taken in the field of chemical safety, we will participate in the large screening effort to prioritize chemicals for further risk assessment. The ultimate impact of this project is o contribute to protection of the population from exposure to obesogens and diabetogens during critical periods in life, which is in line with the overall NIH mission to protect human health through prevention of disease, to protect the developing fetus, and to increase the knowledge of biological effects of environmental contaminants (NIH Mission; ://www.nih.gov/about/mission.htm). In addition, this proposal explores the zebrafish model as a tool for toxicity screening, which is supported by NIH's strong advocacy of zebrafish and other animal model systems for biomedical and behavior research (Henken DB, Rasooly RS, Javois L, Hewitt AT. 2004 National Institutes of Health Trans-NIH Zebrafish Coordinating Committee. The National Institutes of Health and the growth of the zebrafish as an experimental model organism. Zebrafish. 1(2):105-10). ! PUBLIC HEALTH RELEVANCE: Obesity and obesity-related disorders, such as type 2 diabetes, hypertension, and cardiovascular disease, have increased dramatically in Western countries, particularly the United States during the past decades. One emerging cause of the increase might be that we are exposed to industrial chemicals that promote obesity. However, out of the tens of thousands of chemicals that we are exposed to, very few have been tested for their capacity to cause obesity or diabetes, largely due to the lack of models to screen these compounds. In this project, we will produce a screening model to identify chemicals that induce obesity and diabetes. The screening model is based upon zebrafish, which, despite their apparent difference to humans, have similar biological ways of making and storing fat, and producing insulin. These models will be used to screen chemicals and to predict the risk of chemical exposure to humans. !