Human Intestinal Enteroids as Ex Vivo Models of Human Rotavirus Infection

人肠肠类作为人轮状病毒感染的离体模型

• 批准号: 9031047
• 负责人: Mary Kolb Estes
• 金额: $ 27.19万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2020-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9031047
• 关键词: 3-Dimensional; Address; Affect; Africa; Age; Animal Model; Animals; Antigen Receptors; Antigens; Attention; Autophagocytosis; Biocompatible Materials; Cells; Cessation of life; Child; Complex; Country; Cultured Cells; Developing Countries; Disease; Engineering; Enteral; Environment; Epithelial; Ethnic group; Evaluation; Fluids and Secretions; Functional disorder; Gastrointestinal tract structure; Genetic; Growth; High Prevalence; Housing; Human; Immune response; Immunity; Infection; Intestines; Knowledge; Lead; Licensing; Life; Link; Lipids; Mediating; Microbe; Modeling; Molecular; Mus; Organism; Outcome; Pathogenesis; Pathway interactions; Physiological; Play; Population; Predisposition; Prevalence; Probiotics; Property; Resolution; Role; Rotavirus; Rotavirus Infections; Rotavirus Vaccines; Rotavirus disease; Signal Transduction; Stretching; Structure; System; Thinking; Time; Up-Regulation; Vaccination; Vaccines; Viral Pathogenesis; Viral Proteins; Virus Diseases; Vomiting; Work; base; co-infection; cytokine; enteric pathogen; extracellular; gastrointestinal infection; immunogenicity; improved; microbial; microbiome; novel; pathogen; pathogenic Escherichia coli; pathogenic bacteria; permissiveness; prevent; receptor; response; sugar; transmission process; vaccine efficacy

PROJECT SUMMARY Rotavirus (RV) is the leading cause of life-threatening diarrheal disease in children under the age of 5, and results in nearly half a million deaths annually, despite the use of two licensed RV vaccines. Although RV vaccines are available, they do not work optimally in countries where they are needed most. Human rotaviruses (HRVs) grow poorly in cultured cells and do not infect mice, which are only susceptible to animal rotavirus infection. We do not understand how HRV infections cause life-threatening disease and question if knowledge from existing animal models is fully applicable to HRVs. We pioneered establishing human intestinal enteroids (HIEs) as novel ex vivo model “mini-gut” systems to examine host-pathogen interactions and pathophysiology. HIEs are self-organizing, 3-dimensional physiologically active, multicellular cultures that mimic the ordered structure of the intestine. We have found that HRV strains replicate extremely well in HIEs mimicking the host permissiveness observed in people, thus establishing a new model to study HRV infections and pathogenesis. We propose studies to directly examine mechanisms of pathophysiology and genetic control of HRV infection and growth. The recent discovery that cellular receptors used by HRV strains are histoblood group antigens (HBGAs) resulted in a paradigm shift in thinking about the host's genetic influence on RV infection and provides a new perspective on zoonotic transmission of rotaviruses. Unique differences in HBGA expression may explain the higher prevalence of unusual HRV strains in Africa and lower vaccine efficacy seen in distinct ethnic populations. We hypothesize that HRV infection of enteroids is genetically restricted and permissive infection leads to novel intracellular and extracellular innate signaling and response pathways that underlie HRV pathogenesis and pathophysiology. In addition, the outcome of HRV infection or vaccination can be modulated by beneficial microbes. We propose studies in Aim 1 to determine whether genetically-controlled modulators of susceptibility (HBGAs) and distinct innate responses in HIEs regulate HRV infection and pathophysiology (with Cores B and C). Studies in Aim 2 will establish the effect of commensal or pathogenic organisms on HRV or vaccine infection. These studies will elucidate the molecular basis for strain- specific host range restriction observed in children of different ethnic groups and be relevant to understanding vaccine properties. Furthermore, evaluation of modified HIE cultures that house human commensals, probiotics or other pathogens (with Cores B, C and Project 2) in new tailored biomaterial platforms (with Project 3) will lead to a better understanding of how epithelial responses to a viral infection are modulated. Collectively, this work will provide a new understanding of rotavirus pathophysiology and the subsequent critical adaptive responses in humans that trigger immunity and disease.

项目摘要 轮状病毒（RV）是5岁以下儿童威胁生命的腹泻病的主要原因， 每年导致近50万人死亡，使用两种持牌RV疫苗。虽然RV 可以使用疫苗，它们在最需要的国家 /地区无法最佳地工作。人类 轮状病毒（HRV）在培养的细胞中生长不佳，不感染小鼠，而小鼠仅易于动物 轮状病毒感染。我们不了解HRV感染如何引起威胁生命的疾病，并质疑是否是否 现有动物模型的知识完全适用于HRV。我们开创了建立人 肠内肠to（hies）作为新型的离体模型“迷你态”系统，以检查宿主病原体相互作用 和病理生理学。 hies是自组织的，三维体育活动的多细胞培养物 模仿肠的有序结构。我们发现HRV菌株在HIE中复制得很好 模仿在人们中观察到的宿主允许性，从而建立了一种研究HRV感染的新模型 和发病机理。我们提出的研究是直接检查病理生理学和遗传学的机制 控制HRV感染和生长。最近发现，HRV菌株使用的细胞接收器是 Histoblood组抗原（HBGAS）在思考宿主的遗传影响方面导致了范式转变 关于RV感染，并提供了关于轮状病毒的人畜共动性传播的新观点。独特的差异 HBGA表达可能解释了非洲异常HRV菌株和较低疫苗的较高患病率 在不同的种族人群中看到的疗效。我们假设HRV感染了肠to的感染 受限制和允许的感染导致细胞内和细胞外的先天信号传导和反应 HRV发病机理和病理生理学的途径。另外，HRV感染的结果或 可以通过有益的微生物调节疫苗接种。我们建议在目标1中进行研究，以确定是否 易感性（HBGA）的遗传控制调节剂和HIS中不同的先天反应调节HRV 感染和病理生理（核B和C）。 AIM 2中的研究将确定共生或 HRV或疫苗感染的致病生物。这些研究将阐明菌株的分子基础 在不同种族的儿童中观察到的特定寄宿范围限制，与理解有关 疫苗特性。此外，对容纳人类儿童的修改的HIE文化的评估， 益生菌或其他病原体（带有核心B，C和项目2） 项目3）将更好地了解如何调节对病毒感染的上皮反应。 总的来说，这项工作将提供对轮状病毒病理生理学和随后的新理解 引发免疫力和疾病的人类的关键适应性反应。