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（由适用提供）：直到最近，试图鉴定病毒病原体的研究仅限于通过细胞培养，PCR，用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。因此，测试仅限于已经表征的病毒，这在很大程度上是通过对体外细胞的细胞病变作用发现的。现在引入了高通量DNA测序，可以识别出所有病毒的宏基因组方法，并揭示了大量以前未知的病毒，属于许多不同的病毒家族，尤其是在人类以及野生和驯养的动物中。核酸测试的非常高的灵敏度也表明，不仅在腹泻中，而且在健康的人类和动物中，肠道病毒感染率很高。这里将使用一种宏基因组学/生物信息学方法来首先生成与特发性慢性腹泻（ICD）的大量恒河猕猴中所有肠道病毒的全面清单，这是类似于人类溃疡性的结肠的常见炎症状况。为了确定与这种意外疾病相关的候选病毒病原体，经典的病例控制研究将比较大量具有ICD与健康对照的动物中所有病毒的存在和病毒浓度。然后将使用原位杂交测试这些病毒在组织病理学的解剖部位中这些病毒的病毒复制。然后，在最大数量的动物中与ICD最密切相关的顶级候选病毒病原体将口服被口服成少年猕猴，并在结肠中密切监测的ICD，病毒脱落和病毒复制的发展。因此，我们将应用宏基因组学和生物信息学工具的力量，并结合经典的流行病学研究，以对病毒在影响恒河猴猕猴研究殖民地的常见和慢性原发性疾病中的作用进行全包测试。兽医科学专家及其病毒核酸将仔细选择大量动物样品，然后全面分析动物接种以鉴定一种或多种病毒病原体。这项研究还将提高我们对大型私人人群病毒多样性的理解，并证明宏基因组学识别出所有用于随后疾病关联研究的病毒，以比较受影响病例与健康对照的病毒复制。引起的ICD的病毒鉴定不仅将有助于管理生物医学研究中最常使用的非人类灵长类动物的健康，而且还将为病毒诱导的慢性腹泻/结肠炎提供模型系统，以及用于控制灵长类动物中心死亡率和动物变异性的主要来源的疫苗发育。