Viral etiology of idiopathic chronic diarrhea in rhesus macaques

恒河猴特发性慢性腹泻的病毒病因学

基本信息

批准号：
9083165
负责人：
ERIC L DELWART
金额：
$ 52.71万
依托单位：
VITALANT
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：
2016-08-12 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9083165
关键词：
Adenoviruses Adolescent Affect Animals Antibiotic Therapy Antibodies Antigens Autopsy Base Sequence Bioinformatics Biological Assay Biological Models Biomedical Research Body Weight decreased California Case-Control Studies Cell Culture Techniques Cells Chronic Colitis Colon Complex Control Animal DNA DNA Sequence DNA Viruses Dehydration Deposition Development Diagnostic Diarrhea Disease Domestic Animals Enteral Enterovirus Epidemiologic Studies Etiology Family Feces Future Genbank Genome Genomics Health High-Throughput DNA Sequencing Human In Situ Hybridization In Vitro Incidence Infant Inflammatory Laboratories Libraries Life Location Macaca Macaca mulatta Measures Medical Metagenomics Modeling Monitor Neurologic Nucleic Acid Amplification Tests Nucleic Acids Parvovirus Pathogenicity Pathology Performance Picobirnavirus Population Primate Diseases Primates Protocols documentation RNA RNA Viruses Recurrence Reovirus Research Resistance Role Sampling Satellite Viruses Site Source Staining method Stains Testing Time Tissues Ulcerative Colitis Viral Viral Genome Viral Load result Virus Virus Diseases Virus Replication Virus Shedding Wild Animals animal mortality animal tissue base case control cohort deep sequencing design genome sequencing improved in vivo interdisciplinary approach juvenile animal mortality multidisciplinary next generation sequencing nonhuman primate novel novel virus particle pathogen pathogenic bacteria public health relevance respiratory tissue fixing tool vaccine development veterinary science virome virus identification

项目摘要

DESCRIPTION (provided by applicant): Until recently, studies attempting to identify viral pathogens were restricted to testing for previously known viruses detectable by cell culture, PCR, staining with antigen-specific antibodies, or through sero-conversion. Testing was therefore restricted to already characterized viruses, largely those that had been discovered through their cytopathic effects on cells grown in vitro. The introduction of high-throughput DNA sequencing now allows metagenomic approaches to identify all the viruses present and has revealed a large number of previously unknown viruses, belonging to many different viral families, particularly in the feces of humans as well as in wild and domesticated animals. The very high sensitivity of nucleic acid testing has also revealed a high rate of enteric viral infections not just in diarrheic but also in healthy humans and animals. Here a metagenomics/bioinformatics approach will be used to first generate a comprehensive list of all the enteric viruses present in a large cohort of rhesus macaques with idiopathic chronic diarrhea (ICD), a common inflammatory condition of the colon resembling human ulcerative colitis. In order to identify candidate viral pathogens associated with this unexplained disease, a classic case control study will compare the presence and viral concentration of all the virus present in a large number of animals with ICD versus healthy controls. Viral replication of these viruses in the anatomical site of tissue pathology in animals affected with ICD will then be tested using in situ hybridization. The top candidate viral pathogens, most strongly associated with ICD in the largest number of animals, will then be orally inoculated into juvenile macaques and the development of ICD, viral shedding, and viral replication in the colon closely monitored. We will therefore apply the power of metagenomics and bioinformatics tools combined with a classic epidemiological study for an all-inclusive test of the role of viruses in a common and chronic primate disease affecting rhesus macaque research colonies. A large numbers of animal samples will be carefully selected by veterinary science experts and their viral nucleic acids comprehensively analyzed followed by animal inoculations to identify one or more viral pathogens. This study will also advance our understanding of viral diversity in a large primate population, and demonstrate the utility of metagenomics to identify all viruses present for subsequent disease association studies comparing viral replication in affected cases versus healthy controls. The identification of virus(es) causing ICD will not only assist managing the health of the non-human primate species most often used in biomedical research but will also provide model systems for virally induced chronic diarrhea/colitis and viral targets for vaccine development to control a major source of mortality and animal variability in primate centers.

描述（由申请人提供）：直到最近，试图鉴定病毒病原体的研究仅限于测试可通过细胞培养、PCR、抗原特异性抗体染色或通过血清转化检测到的先前已知病毒。高通量 DNA 测序的引入现在允许宏基因组方法识别所有存在的病毒，并揭示了大量以前未知的病毒，这些病毒属于病毒。许多不同的病毒家族，特别是在人类以及野生和家养动物的粪便中，核酸检测的极高灵敏度也表明，不仅在腹泻患者中，而且在健康的人类和动物中，肠道病毒感染率也很高。这里将使用宏基因组学/生物信息学方法首先生成一大群患有特发性慢性腹泻（ICD）的恒河猴中存在的所有肠道病毒的综合列表，ICD是一种常见的炎症性疾病。为了识别与这种无法解释的疾病相关的候选病毒病原体，一项经典的病例对照研究将比较大量 ICD 动物和健康对照动物中存在的所有病毒的存在情况和病毒浓度。然后，将使用原位杂交测试这些病毒在受 ICD 影响的动物的组织病理学解剖部位中的复制，然后将口服接种与 ICD 最密切相关的顶级候选病毒病原体。因此，我们将利用宏基因组学和生物信息学工具，结合经典的流行病学研究，对病毒在结肠中的作用进行全面测试。一种影响恒河猴研究群体的常见慢性灵长类疾病，兽医科学专家将精心挑选大量动物样本，对其病毒核酸进行全面分析，然后对动物进行接种以进行鉴定。这项研究还将增进我们对大型灵长类动物群体中病毒多样性的了解，并证明宏基因组学可用于识别所有病毒，以便进行后续疾病关联研究，比较受影响病例与健康对照中的病毒复制情况。引起 ICD 的病毒不仅有助于生物医学研究中最常使用的非人类灵长类动物的健康，而且还将提供病毒引起的慢性腹泻/结肠炎的模型系统和用于疫苗开发以控制主要来源的病毒靶点死亡率和动物变异性灵长类动物中心。