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， 每年直接用于医疗系统的费用高达10亿美元。由于潜在的免疫抑制和缺乏有效的 在抗病毒治疗中，移植患者可能会因HuNoV感染而产生严重的后遗症。虽然 细菌肠道微生物群已显示在临床前增强肠道病毒的复制和致病性， 在HuNoV感染模型中，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，我们的目标是获得更大的机械 深入了解肠道微生物群如何调节移植患者的HuNoV感染。我们的核心假设是， 具有症状性HuNoV感染的移植患者将具有肠道微生物组特征，其显示 富集促进HIE感染的特定肠道微生物群（肠杆菌科）， 从HuNoV感染发展慢性症状将伴随特定肠道微生物群的消耗 调节宿主先天性免疫反应（1型干扰素）。在目标1中，我们将进一步定义两种肠道 移植患者±HuNoV感染的微生物组和宿主因子差异。首先，我们将建立一个更大的 成人和儿童移植患者队列并收集纵向粪便标本。然后，我们将执行 全面的肠道微生物组分析（宏基因组鸟枪测序和细菌qPCR），以确认我们的 初步结果。我们预测，HuNoV感染的移植患者将具有显著不同的肠道 与未感染的对应物相比，微生物群特征，以及更高强度的免疫抑制和 高抗生素负荷将对应于HuNoV感染移植患者的慢性腹泻。在目标2中，我们将 使用体外HIE模型确定肠杆菌科是否促进HuNoV感染。首先，我们会感染空肠 将HIE与HuNoV共孵育，并将HuNoV与各种肠杆菌科物种共孵育。然后，我们将执行RNA 提取物和定量HuNoV RT-qPCR以评估共孵育各种肠杆菌科的效果 spp.关于病毒复制。我们预测肠杆菌科促进HIEs的HuNoV感染。如果是这种 在这种情况下，我们将研究这是否是一种仅在HBGA表达成员中观察到的现象。 肠杆菌科在目标3中，我们将确定患有慢性症状性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系统性细胞因子特征。我们将进行批量细胞因子 使用Isoplexis分析血清样品，Isoplexis是一种新型功能蛋白质组学分析平台。我们预测 患有由HuNoV感染引起的慢性腹泻的移植患者将表现出类杆菌属的缺乏， 导致1型干扰素耗竭，然后导致Th 1免疫应答降低， Th 2免疫应答和Th 2优势细胞因子（IL-4、IL-5、IL-10）表达增加。拟议 实验，培训和教学课程，在这个K23将装备候选人（钟博士）独特的 技能，使她能够过渡到独立作为一个医生科学家在肠道微生物-HuNoV 移植患者的互动。