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.

项目概要 人类诺如病毒 (HuNoV) 是美国急性胃肠炎的主要原因，造成 4.2 美元的损失 每年 10 亿美元的直接卫生系统费用。由于潜在的免疫抑制和缺乏有效的 如果接受抗病毒治疗，移植患者可能会因 HuNoV 感染而出现严重的后遗症。尽管 临床前研究表明，细菌肠道微生物群可增强肠道病毒的复制和发病机制 模型中，其在 HuNoV 感染中的作用仍然很大程度上未知。在这个项目中，我们的目标是获得更大的机械性 深入了解肠道微生物群如何调节移植患者的 HuNoV 感染。我们的中心假设是 患有 HuNoV 感染症状的移植患者的肠道微生物组特征显示 促进 HIE 感染的特定肠道微生物群（肠杆菌科）的富集，以及那些 HuNoV 感染出现慢性症状将伴随特定肠道微生物群的耗竭 调节宿主先天免疫反应（1 型干扰素）。在目标 1 中，我们将进一步定义肠道 移植患者的微生物组和宿主因素差异±HuNoV 感染。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（宏基因组鸟枪法测序和细菌 qPCR）以确认我们的研究 初步结果。我们预测 HuNoV 感染的移植患者的肠道会有显着不同 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 高抗生素负荷将与 HuNoV 感染的移植患者的慢性腹泻相对应。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，我们要感染空肠 HIE 与 HuNoV 以及 HuNoV 与各种肠杆菌科细菌共同培养。然后，我们将进行 RNA 提取和定量 HuNoV RT-qPCR 评估共培养各种肠杆菌科细菌的效果 种。关于病毒复制。我们预测肠杆菌科促进 HIE 的 HuNoV 感染。如果这是 情况下，我们将调查这是否是仅在 HBGA 表达成员中观察到的现象 肠杆菌科。在目标 3 中，我们将确定移植患者是否患有慢性症状性 HuNoV 感染 与未感染的对应物相比，具有不同的全身细胞因子特征。我们将进行批量细胞因子 利用 Isoplexis（一种新型功能蛋白质组分析平台）对血清样品进行分析。我们预测 因 HuNoV 感染而患有慢性腹泻的移植患者将表现出拟杆菌属的缺乏， 导致 1 型干扰素耗竭，进而导致 Th1 免疫反应降低、增加 Th2 免疫反应和 Th2 主要细胞因子（IL-4、IL-5、IL-10）表达增加。拟议的 K23 中的实验、培训和教学课程将为候选人（Chong 博士）提供独特的能力 将使她能够独立成为肠道菌群医学科学家的技能-HuNoV 移植患者之间的相互作用。