Uncovering small RNAs that contribute to Coxiella burnetii infection

发现导致伯内氏柯克斯体感染的小 RNA

基本信息

批准号：
9529169
负责人：
Rahul Raghavan
金额：
$ 7.43万
依托单位：
PORTLAND STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-19 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9529169
关键词：
Acute Aerosols Affect Anaplasma Bacteria Binding Bioinformatics Biological Assay Biology Cell physiology Cells Chlamydia Chronic Coxiella Coxiella burnetii Data Development Disease Outbreaks Distant Ehrlichia Endocarditis Enterobacteriaceae Environment Epidemic Etiology Gene Expression Gene Expression Regulation Genes Genus staphylococcus Goals Goat Growth Human Infection Knowledge Lead Legionella Livestock Measures Mediating Membrane Proteins Messenger RNA Mitochondria Molecular Netherlands Northern Blotting Organism Outcome Pathogenesis Pathogenicity Process Proteins Public Health Q Fever Quantitative Reverse Transcriptase PCR Regulation Regulator Genes Regulon Research Ribonucleases Rickettsia Role Small RNA Technology Testing Time Translations Treatment Efficacy Untranslated RNA Validation Virulence Work Zoonoses base improved innovation insight macrophage monocyte mutant novel novel therapeutic intervention novel therapeutics pathogen prevent targeted treatment transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT Non-coding small RNAs (sRNAs), which act swiftly and specifically, are ideal gene regulators in pathogens such as Coxiella burnetii that shift rapidly between disparate environments (e.g. aerosol vs. intracellular). However, very little is known about sRNAs that facilitate the pathogenicity of C. burnetii—a select agent that causes acute Q fever and chronic endocarditis. It is important that we gain better molecular insights into C. burnetii infection because it is found across the globe and has the potential to cause epidemics such as the recent outbreak in the Netherlands that involved more than 4,000 human cases and resulted in the culling of over 50,000 goats (a primary reservoir). Our long-term goal is to define the functions of non- coding RNAs during C. burnetii infection and to apply this knowledge to developing novel therapeutic ap- proaches. Towards attaining this goal, the objective of this application is to identify sRNAs that promote C. burnetii's growth within human cells. Based on preliminary data that identified several novel sRNAs in Cox- iella, our central hypothesis is that sRNAs promote intracellular growth of C. burnetii. The objectives of this project will be accomplished by two Specific Aims: (1) Identify sRNAs that are important to Coxiella's intra- cellular growth. Using RNA-seq we expect to identify sRNAs that are expressed during various stages of in- fection, and by examining transposon-insertion mutants we will assay their importance to intracellular growth. (2) Define the role of the sRNA CbsR14 in promoting Coxiella's intracellular growth. To begin to understand how sRNAs facilitate infection, we will interrogate the function of CbsR14, This sRNA was cho- sen because its expression was induced intracellularly, a transposon insertion in it significantly reduced Coxiella's intracellular growth, and preliminary analyses indicated that it targets the adjacent yidC gene. All studies to date on C. burnetii infection have only investigated proteins; hence, the research proposed here is innovative because it will depart from the status quo to investigate the role of non-coding RNAs in facilitat- ing Coxiella infection. This study is significant because it will, for the first time, identify sRNAs that pro- mote intracellular growth of C. burnetii. This new knowledge is expected to advance the field considerably by transforming our current understanding of Coxiella's pathogenicity. Additionally, uncovering the regula- tory circuit of YidC could be applicable to other organisms because this membrane protein is conserved across bacteria and mitochondria. Furthermore, the approaches we develop in this project will guide us (and others) to analyze other classes of non-coding RNAs (e.g. riboswitches) in Coxiella, and to examine sRNAs in other intractable intracellular pathogens such as Chlamydia, Rickettsia, Anaplasma and Ehrlichia. Moreover, a deeper understanding of sRNA-based gene regulation could contribute to the development of novel anti-Coxiella therapeutic agents that target critical components of the sRNA regulon.

项目摘要/摘要非编码小RNA（SRNA），迅速，具体地起作用，是病原体中理想的基因调节剂例如coxiella burnetii在不同的环境（例如气溶胶与细胞内）之间迅速转移。但是，关于促进C. burnetii的致病性的SRNA知之甚少这会导致急性Q热和慢性心内膜炎。重要的是要获得更好的分子见解进入C. burnetii感染，因为它在全球范围内发现，并且有可能引起发作作为最近在荷兰的爆发，涉及4,000多个人类案件，并导致淘汰超过50,000山羊（主要储层）。我们的长期目标是定义非 - 在C. burnetii感染过程中编码RNA，并将这些知识应用于开发新的治疗性AP- 促进。为了实现这一目标，本应用的目的是确定促进C的SRNA。 Burnetii在人类细胞中的生长。基于初步数据，该数据鉴定了Cox-中的几个新型SRNA IELLA，我们的中心假设是SRNA促进了burnetii的细胞内生长。目标的目标项目将通过两个具体目标来完成：（1）确定对Coxiella内部至关重要的SRNA 细胞生长。使用RNA-seq，我们期望识别在内部各个阶段表达的SRNA 派系，通过检查转座子插入突变体，我们将主张它们对细胞内的重要性（2）定义了SRNA CBSR14在促进Coxiella细胞内生长中的作用。开始了解SRNA如何促进感染，我们将询问CBSR14的功能，该SRNA是cho的 SEN由于其表达是细胞内诱导的，因此中插座插入显着降低 Coxiella的细胞内生长和初步分析表明，它靶向相邻的YIDC基因。全部迄今为止，对伯内蒂感染的研究仅研究了蛋白质。因此，这里提出的研究是创新性是因为它将偏离现状，以调查非编码RNA在设施中的作用 coxiella感染。这项研究很重要，因为它将首次确定srnas MOTE C. burnetii的细胞内生长。期望这一新知识会大大推进领域通过改变我们当前对Coxiella致病性的理解。另外，揭示了调节 YIDC的保守回路可能适用于其他生物，因为该膜蛋白是保守的跨细菌和线粒体。此外，我们在该项目中开发的方法将指导我们（以及其他）分析其他类别的非编码RNA（例如核糖开关），并检查SRNA 在其他棘手的细胞内病原体中，例如衣原体，立克，anaplasma和Ehrlichia。此外，对基于SRNA的基因调节的更深入了解可能有助于发展靶向SRNA调节的关键成分的新型抗氧化菌治疗剂。