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.

我们对传染病理解的一个主要差距是缺乏有关分子相互作用的信息感染病原体和人类宿主之间的网络，这对于细菌来说尚未完成该应用程序的目的是在转录水平上定义互作组。使用 RNA 在实验感染的人类志愿者中检测主要皮肤病原体杜克雷嗜血杆菌 (HD) 测序 (RNA-seq) 和代谢组学会导致软下疳，这是一种促进生殖器溃疡 (GU) 的疾病。 HIV 传播是雅司病流行国家儿童皮肤溃疡 (CU) 的主要原因。由于环境储存库，用抗生素根除 HD 相关 CU 的努力失败了。 HD生物学，我们开发了一个模型，其中接种HD GU菌株35000HP及其衍生物全基因组测序表明，大多数 CU 菌株都存在于成年志愿者上臂的皮肤中。与 35000HP 几乎相同，表明我们的模型与 GU 和 CU 高度相关。天然软下疳，HD 存在于脓肿中，周围被多形核白细胞包围，在我们新的初步数据中，我们展示了巨噬细胞并通过逃避吞噬作用保持在细胞外。 HD 和宿主之间存在相互作用网络，并且宿主和细菌转录本存在差异 HD 的表达与感染部位和受伤部位的代谢组变化相关。涉及适应厌氧以及金属离子的吸收和利用的基因的表达替代碳源，例如抗坏血酸，符合“营养毒力”的理念。代谢组学数据显示，抗坏血酸途径在病变中上调。宿主调节其基因转录以吞噬并限制 HD 的营养物质，HD 调节其基因转录来抵消这些宿主防御，宿主基因表达与感染部位的代谢组学特征，以及参与适应环境的细菌基因宿主反应所产生的代谢生态位是毒力所必需的，我们的具体目标是 1) 。定义 HD 与受感染组织中人类宿主之间的代谢组和相互作用组，并将宿主转录反应与病变代谢变化相关联；2) 识别负责的细胞；使用单细胞RNA-seq进行宿主转录反应3)确定HD基因是否存在对人类的毒力需要参与利用宿主生态位以及其机制这些基因有助于毒力，这项研究的重要性在于我们将成为第一个确定的基因。人类细菌感染位点的相互作用组，定义了负责转录的宿主细胞反应，将宿主反应与病变代谢组变化相关联，并研究 HD 如何利用这些变化我们将为模型细胞外细菌与代谢物之间的相互作用提供新的见解。人类宿主并揭示了控制 HD 相关 CU 的新策略。