MicroRNA mediated inhibition of apoptosis in Coxiella burnetii infection

MicroRNA介导的伯内氏柯克斯体感染细胞凋亡抑制

• 批准号: 9243896
• 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
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-15 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243896
• 关键词: Acute; Affect; Anaplasma phagocytophilum; Antibiotics; Apoptosis; Apoptotic; BCL2 gene; Bacteria; Binding; Biological Assay; Biological Process; Biology; Cells; Chlamydia trachomatis; Chronic; Colorectal; Coxiella; Coxiella burnetii; Data; Development; Disease Outbreaks; Endocarditis; Epidemic; Eukaryota; Gene Targeting; Goals; Growth; Human; Infection; Infection Control; Inhibition of Apoptosis; Knowledge; Mediating; Messenger RNA; MicroRNAs; Molecular; Netherlands; Outcome; Pathogenicity; Pathway interactions; Pharmacologic Substance; Play; Process; Proteins; Public Health; Q Fever; Quantitative Reverse Transcriptase PCR; RNA; RNA analysis; Regulation; Research; Role; Testing; Therapeutic; Translations; Type IV Secretion System Pathway; Untranslated RNA; Vacuole; Work; Zoonoses; base; differential expression; improved; innovation; insight; macrophage; monocyte; mortality; novel; overexpression; pathogen; response; targeted agent; transcriptome sequencing

Project Summary/Abstract The Q fever pathogen Coxiella burnetii inhibits apoptosis (programmed cell death) in host cells in order to replicate in a stable intracellular compartment. Effector proteins secreted by Coxiella into host cell have been shown to inhibit apoptosis, although their mechanisms of action are not fully understood. Additionally, no information is currently available about the roles host microRNAs (miRNAs) potentially play in regulating apoptosis in Coxiella-infected cells. miRNAs are ~22 nt RNAs that regulate several biological processes in human cells, including apoptosis, by binding to target mRNAs and either blocking translation or causing target degradation. Our long-term goal is to understand how non-coding RNAs regulate Coxiella-host interactions. Towards attaining this goal, the objective of this application is to identify miRNAs that have a role in inhibiting apoptosis during Coxiella infection. Based on preliminary data that revealed potential functions for apoptosis-related miRNAs in Coxiella-infected macrophages, our central hypothesis is that miRNAs have a role in delaying apoptosis during Coxiella infection. The objective of this project will be accomplished by three specific aims: (1) Identify miRNAs that are induced or repressed by Coxiella infection. Using RNA-seq and qRT-PCR we will identify all miRNAs that are differentially expressed in Coxiella- infected THP-1 (human monocyte/macrophage) cells. (2) Determine miRNAs that inhibit apoptosis during Coxiella infection. For the miRNAs identified in our preliminary studies and in Aim 1, we will assay their ability to inhibit apoptosis. (3) Define the role of miR-148a during Coxiella infection. To begin to understand at a molecular level how miRNAs facilitate inhibition of apoptosis during Coxiella infection, we will interrogate the function of miR-148a. This miRNA was chosen because it was significantly down-regulated in Coxiella-infected cells in our preliminary analysis, and its target gene (Bcl-2) has critical roles in both C. burnetii development and apoptosis inhibition. The research proposed here is innovative, because it will depart from the status quo to investigate a whole new realm of host-pathogen dynamics: the RNA-level interaction between Coxiella and human cells. This work is significant because this will be the first study to uncover the roles of miRNAs during Coxiella infection, and based on our results, novel pharmaceutical agents that target miRNAs could potentially be developed to control chronic Coxiella infections, which are difficult to treat with currently available antibiotics. Our approach and results could also be broadly applied to studying the functions of miRNAs during infections caused by other intracellular pathogens.

项目摘要/摘要 Q发烧病原体Coxiella burnetii抑制宿主细胞中凋亡（程序性细胞死亡） 为了在稳定的细胞内室中复制。 Coxiella分泌的效应蛋白 宿主细胞已显示可抑制细胞凋亡，尽管它们的作用机制尚未完全 理解。此外，目前尚无有关角色主机microRNA的信息 （miRNA）有可能在调节感染的细胞中调节凋亡。 miRNA是〜22 nt RNA 通过与靶标的结合来调节包括凋亡在内的人类细胞中的几个生物学过程 mRNA并阻止翻译或导致目标降解。我们的长期目标是 了解非编码RNA如何调节Coxiella-Host相互作用。为了实现这个目标， 该应用的目的是确定在抑制凋亡中作用的miRNA 考克斯菌感染。基于揭示与凋亡相关的潜在功能的初步数据 我们的中心假设是在柯西埃拉感染的巨噬细胞中的miRNA，是miRNA在 在考克斯菌感染过程中延迟凋亡。该项目的目的将由 三个特定的目的：（1）识别由Coxiella感染诱导或抑制的miRNA。使用 RNA-Seq和QRT-PCR我们将确定所有在Coxiella-中差异表达的miRNA 感染的THP-1（人单核细胞/巨噬细胞）。 （2）确定抑制凋亡的miRNA 在考克斯菌感染期间。对于我们的初步研究和AIM 1中确定的miRNA，我们将 测定它们抑制凋亡的能力。 （3）定义miR-148a在考西菌感染过程中的作用。到 开始在分子水平上了解miRNA如何促进考西ella期间的凋亡抑制 感染，我们将询问miR-148a的功能。选择了这个mirna，因为它是 在我们的初步分析中，在Coxiella感染的细胞中显着下调，其靶基因 （Bcl-2）在弯曲梭菌发育和凋亡抑制中具有关键作用。研究 这里提出的是创新的，因为它将偏离现状来调查一个全新的 宿主 - 病原体动力学的领域：考克斯菌与人类细胞之间的RNA级相互作用。这 工作很重要，因为这将是第一个揭示miRNA在考克斯氏菌中角色的研究 感染，并根据我们的结果，靶向miRNA的新型药物可能有可能 开发以控制慢性考克斯菌感染，这些感染很难治疗当前可用的 抗生素。我们的方法和结果也可以广泛应用于研究的功能 由其他细胞内病原体引起的感染过程中的miRNA。