The role of gut microbiota in human norovirus infections in transplant patients

肠道微生物群在移植患者诺如病毒感染中的作用

• 批准号: 10524718
• 负责人: Pearlie Pao Ee Chong
• 金额: $ 18.55万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-22 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10524718
• 关键词: Acute; Adherent Culture; Adult; Antibiotics; Antiviral Agents; B-Lymphocytes; Bacteroides; Bacteroides fragilis; Binding; Bioinformatics; Blood Group Antigens; CD8-Positive T-Lymphocytes; Cells; Cessation of life; Childhood; Chronic; Chronic diarrhea; Clinical; Clinical Trials; Computerized Medical Record; Data; Dendritic cell activation; Enteral; Enterobacteriaceae; Etiology; Exhibits; Family; Flow Cytometry; Foundations; Future; Gastroenteritis; Goals; Grant; Health system; Hematopoietic Stem Cell Transplantation; Human; Immune response; Immunocompromised Host; Immunology; Immunosuppression; In Vitro; Incubated; Individual; Infection; Inflammatory; Innate Immune Response; Integration Host Factors; Interferon Type I; Interferon-alpha; Interferons; Interleukin-10; Interleukin-4; Interleukin-5; Intestines; Klebsiella; Malnutrition; Mediating; Metagenomics; Modeling; Molecular; Mus; Natural History; Norovirus; Organ; Pathogenesis; Pathogenicity; Patients; Physicians; Play; Polysaccharides; Pre-Clinical Model; Probiotics; Production; Proteins; Proteomics; RNA; Resolution; Role; Sample Size; Sampling; Scientist; Serum; Severities; Shotgun Sequencing; Solid; Surface; T cell response; TNF gene; Taxonomy; Therapeutic; Training; Transplant Recipients; United States; Viral; Viral Pathogenesis; Virus; Virus Replication; Wasting Syndrome; abstracting; cohort; cost; cytokine; enteric virus infection; experimental study; fecal transplantation; gut bacteria; gut microbiome; gut microbiota; human microbiota; insight; member; microbiome signature; microbiota profiles; next generation sequencing; novel; persistent symptom; response; skills; stool sample; tool

PROJECT SUMMARY Human norovirus (HuNoV) is the leading cause of acute gastroenteritis in the United States, resulting in $4.2 billion in direct health system costs annually. Due to underlying immunosuppression and the lack of effective antiviral therapeutics, transplant patients may develop serious sequelae from HuNoV infections. Though bacterial gut microbiota has been shown to enhance replication and pathogenesis of enteric viruses in preclinical models, its role in HuNoV infection remains largely unknown. In this project, we aim to gain greater mechanistic insights into how gut microbiota modulate HuNoV infection in transplant patients. Our central hypothesis is that transplant patients with symptomatic HuNoV infection will have a gut microbiome signature showing an enrichment of specific gut microbiota (Enterobacteriaceae) that facilitate infection of HIEs, and those who develop chronic symptoms from HuNoV infection will have a concomitant depletion of specific gut microbiota that modulate host innate immune responses (type 1 interferons). In Aim 1, we will further define both gut microbiome and host factor differences in transplant patients ±HuNoV infections. First, we will establish a larger cohort of adult and pediatric transplant patients and collect longitudinal stool specimens. Then, we will perform comprehensive gut microbiome profiling (metagenomic shotgun sequencing and bacterial qPCR) to confirm our preliminary results. We predict that HuNoV-infected transplant patients will have significantly different gut microbiota signatures compared to uninfected counterparts, and that higher intensity of immunosuppression and high antibiotic load will correspond with chronic diarrhea in HuNoV-infected transplant patients. In Aim 2, we will determine if Enterobacteriaceae facilitate HuNoV infection using an in vitro HIE model. First, we will infect jejunal HIEs with HuNoV and co-incubate HuNoV with various Enterobacteriaceae spp. Then, we will perform RNA extractions and quantitative HuNoV RT-qPCR to evaluate the effect of co-incubating various Enterobacteriaceae spp. on viral replication. We predict that Enterobacteriaceae promotes HuNoV infection of HIEs. If this is the case, we will investigate if this is a phenomenon observed only in HBGA-expressing members of Enterobacteriaceae. In Aim 3, we will determine if transplant patients with chronic symptomatic HuNoV infections have different systemic cytokine signatures compared to uninfected counterparts. We will perform bulk cytokine analysis on serum samples utilizing Isoplexis, a novel functional proteomic profiling platform. We predict that transplant patients with chronic diarrhea from HuNoV infection will exhibit a paucity of genus Bacteroides, resulting in depletion of type 1 interferons, which then leads to a decreased Th1 immune response, increased Th2 immune response and increased expression of Th2-predominant cytokines (IL-4, IL-5, IL-10). The proposed experiments, training and didactic coursework in this K23 will equip the candidate (Dr. Chong) with unique skillsets that will enable her transition to independence as a physician scientist in gut microbiota-HuNoV interactions in transplant patients.

项目摘要 人类诺如病毒（Hunov）是美国急性胃肠炎的主要原因，导致4.2美元 每年有十亿个直接卫生系统成本。由于潜在的免疫抑制和缺乏有效的 抗病毒疗法，移植患者可能会因匈奴感染而出现严重的后遗症。尽管 细菌肠道菌群已显示可增强临床前肠道病毒的复制和发病机理 模型，其在Hunov感染中的作用在很大程度上仍然未知。在这个项目中，我们旨在获得更大的机械 洞察肠道微生物群如何调节移植患者的Hunov感染。我们的中心假设是 有症状的Hunov感染的移植患者将具有肠道微生物组签名，显示 富含促进HIE感染的特定肠道菌群（肠杆菌科）和 霍诺夫感染产生慢性症状将伴随特定肠道菌群的耗竭 该调节宿主先天免疫反应（1型干扰素）。在AIM 1中，我们将进一步定义两个肠道 移植患者的微生物组和宿主因子差异±Hunov感染。首先，我们将建立更大的 成人和小儿移植患者的队列并收集纵向粪便标本。然后，我们将表演 全面的肠道微生物组分析（宏基因组shot弹枪测序和细菌QPCR）确认我们 初步结果。我们预测，饥饿感染的移植患者的肠道明显不同 与未感染的对应物相比，微生物群的特征，并且免疫抑制强度更高 高抗生素负荷将与饥饿感染的移植患者中的慢性腹泻相对应。在AIM 2中，我们将 确定使用体外HIE模型的肠杆菌科是否喜欢的Hunov感染。首先，我们将感染的空肠 HIE与Hunov和Hunov共同进行了各种肠杆菌科。然后，我们将执行RNA 提取和定量Hunov rt-QPCR，以评估共同结合各种肠杆菌科的效果 spp。关于病毒复制。我们预测，肠杆菌科会促进HIE的Hunov感染。如果这是 情况，我们将调查是否仅在表达HBGA的成员中观察到这是一种现象 肠杆菌科。在AIM 3中，我们将确定患有慢性症状Hunov感染的移植患者是否 与未感染的对应物相比，具有不同的全身细胞因子特征。我们将执行散装细胞因子 使用等形的血清样品分析，这是一种新型的功能性蛋白质组学分析平台。我们预测 霍诺夫感染患有慢性腹泻的移植患者将表现出很少的杀菌属， 导致1型中断的耗竭，然后导致Th1免疫反应下降，增加 Th2免疫响应和Th2-抗抑制细胞因子（IL-4，IL-5，IL-10）的表达增加。提议 该K23中的实验，培训和教学课程将为候选人（Chong博士）配备独特 能够在肠道微生物群中成为物理科学家过​​渡到独立性的技能集 移植患者的相互作用。