Determination of the Interactome between Haemophilus ducreyi and the Human Host.

杜克雷嗜血杆菌与人类宿主之间相互作用组的测定。

基本信息

批准号：
10531548
负责人：
Stanley M. Spinola
金额：
$ 58.04万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-17 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10531548
关键词：
Abscess Adult Affect Africa Anaerobiosis Annual Reports Antibiotics Ascorbic Acid Asia Azithromycin Bacteria Bacterial Genes Bacterial Infections Binding Biology Carbon Case Study Cells Chancroids Child Chronic Communicable Diseases Country Cutaneous Data Development Disease Endemic Diseases Gene Expression Genes Genetic Transcription Genital Genitalia Growth HIV HIV-1 Hemophilus ducreyi Host Defense Human Human Volunteers Immune response In Vitro Individual Infection Infectious Skin Diseases Ions Leprosy Lesion Macrophage Metabolic Metals Micro Array Data Modeling Molecular Mus Nutrient Nutritional Organism Papua New Guinea Pathogenesis Pathway interactions Phagocytes Phagocytosis Pharmaceutical Preparations Publications Site Skin Skin colonization Source Testing Tissues Transcript Treponema pallidum Ulcer Upper arm Vaccines Virulence World Health Organization Yaws case finding disability extracellular fly genome sequencing in vivo infection risk insight metabolome metabolomics neglected tropical diseases neutrophil novel strategies pathogen pressure response single-cell RNA sequencing skin disorder therapeutic development transcriptome sequencing transmission process uptake volunteer whole genome

项目摘要

A major gap in our understanding of infectious diseases is the lack of information about molecular interaction networks between an infecting pathogen and the human host, which has yet to be accomplished for a bacterial infection in humans. The purpose of this application is to define an interactome on a transcript level for the primary skin pathogen, Haemophilus ducreyi (HD), in experimentally infected human volunteers using RNA sequencing (RNA-seq) and metabolomics. HD causes chancroid, a genital ulcer (GU) disease that facilitates HIV transmission and is as a major cause of cutaneous ulcers (CU) in children in yaws-endemic countries. Efforts to eradicate HD-associated CU with antibiotics failed due to environmental reservoirs. To study the biology of HD, we developed a model in which the HD GU strain 35000HP and its derivatives are inoculated into the skin of the upper arm of adult volunteers. Whole genome sequencing shows that most CU strains are nearly identical to 35000HP, indicating that our model is highly relevant to GU and CU. In the model and in natural chancroid, HD resides in an abscess where it is surrounded by polymorphonuclear leukocytes and macrophages and remains extracellular by evading phagocytosis. In our new preliminary data, we show that an interaction network exists between HD and the host and that differential host and bacterial transcript expression correlates with metabolomic changes at infected vs. wounded sites. HD primarily upregulates the expression of genes involved in adapting to anaerobiosis and uptake and utilization of metal ions and alternative carbon sources, such as ascorbic acid, consistent with the idea of “nutritional virulence.” Our new metabolomics data show that ascorbic acid pathways are upregulated in lesions. Thus, we hypothesize that the host regulates its gene transcription to phagocytize and limit nutrients to HD, that HD regulates its gene transcription to counteract these host defenses, that host gene expression correlates with metabolomic profiles at infected sites, and that bacterial genes that are involved in adaptation to the metabolic niche created by the host response will be required for virulence. Our specific aims are 1) to define the metabolome and the interactome between HD and the human host in infected tissue and to correlate the host transcriptional response to metabolic changes in lesions; 2) to identify the cells responsible for the host transcriptional response using single cell RNA-seq; 3) to determine whether the HD genes involved in exploitation of the host niche are required for virulence in humans and the mechanism(s) by which these genes contribute to virulence. The importance of this study is that we will be the first to determine an interactome at a site of a bacterial human infection, define the host cells responsible for the transcriptional response, correlate the host response to metabolomic changes in lesions, and study how HD exploits these metabolites. We will provide new insights into the interaction between a model extracellular bacterium and the human host and unmask novel strategies to control HD-associated CU.

我们对传染病的理解的主要差距是缺乏有关分子相互作用的信息受感染的病原体与人类宿主之间的网络，尚未完成细菌人类感染。此应用程序的目的是在转录级上定义一个互动组在实验感染的人类志愿者中，原发性皮肤病原体，嗜血杆菌Ducreyi（HD）测序（RNA-SEQ）和代谢组学。高清导致chancroid，一种促进的生殖器溃疡（GU）疾病艾滋病毒传播是偏航国家儿童皮肤溃疡（CU）的主要原因。由于环境储层而导致的放射性HD相关铜与抗生素相关的CU失败。研究 HD的生物学，我们开发了一个模型，其中HD GU菌株35000HP及其衍生物被接种进入成年志愿者上臂的皮肤。整个基因组测序表明大多数Cu菌株是几乎与35000HP相同，表明我们的模型与GU和CU高度相关。在模型和天然chancroid，HD居住在脓肿中，在该脓肿中被多形核白细胞包围巨噬细胞通过逃避吞噬作用，保持细胞外。在我们的新初步数据中，我们显示高清和宿主之间存在一个相互作用网络以及差异宿主和细菌转录本表达与受伤部位与受伤部位的代谢组变化相关。高清主要上调与适应厌氧菌病的基因表达，金属离子的摄取和利用和利用替代碳源，例如抗坏血酸，与“营养病毒”的概念一致。我们的新代谢组学数据表明，抗坏血酸途径在病变中已更新。那我们假设宿主调节其基因转录以吞噬细胞并将养分限制在HD中，该HD调节其调节基因转录以抵消这些宿主防御，宿主基因表达与在受感染部位的代谢组谱，以及与适应的细菌基因病毒需要由宿主反应产生的代谢生态市场。我们的具体目标是1）在受感染组织中定义HD和人类宿主之间的代谢组和相互作用组将宿主转录反应与病变的代谢变化相关联； 2）识别负责的单元用于使用单细胞RNA-Seq的宿主转录响应； 3）确定HD基因是否参与人类病毒需要剥削宿主利基和宿主利基市场需要这些基因有助于病毒。这项研究的重要性是，我们将是第一个确定一个在人类感染的部位相互作用，定义负责转录的宿主细胞反应，将宿主反应与病变的代谢组变化相关，并研究HD如何利用这些。代谢物。我们将提供有关模型细胞外细菌与该模型之间相互作用的新见解人类宿主并揭露了控制HD相关CU的新颖策略。