Investigating the extracellular RNA content of Staphylococcus aureus

金黄色葡萄球菌胞外 RNA 含量的研究

• 批准号: 10289001
• 负责人: Ronan Carroll
• 金额: $ 7.55万
• 依托单位: OHIO UNIVERSITY ATHENS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-13 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10289001
• 关键词: Architecture; Bacteria; Bacterial Adhesins; Cell Wall; Cells; DNA; Dangerousness; Data; Future; Gram-Positive Bacteria; Growth; Human; Human body; Immune; Immune Evasion; Immune response; Immune system; Infection; Inflammatory; Interferon Type I; Knowledge; Life; Lipids; Membrane; Nucleic Acids; Pathogenesis; Pathogenicity; Play; Production; Property; Proteins; RNA; Reporting; Research; Role; Source; Staphylococcus aureus; Staphylococcus aureus infection; Superantigens; Time; Toll-like receptors; Toxin; Vesicle; Virulence; Virulence Factors; cytokine; cytotoxic; exoenzyme; extracellular; extracellular vesicles; immune activation; microbial; nanosized; pathogenic bacteria; stem cells; transcriptome sequencing; vesicular release; virtual

PROJECT SUMMARY/ABSTRACT Extracellular vesicles (EVs) are nano sized, membrane-enclosed vesicles produced and released by cells in all domains of life. EVs contain a variety of biomolecules, such as lipids, proteins, and nucleic acids, and are involved in a broad range of functions depending on the progenitor cell. The study of EVs in Gram-positive bacteria is a relatively new field. Due to the architecture of the Gram-positive cell wall it was long thought to be impossible for Gram-positive bacteria to produce EVs. However recent studies have shown EV production in a variety of Gram-positive bacteria and implicated EVs in important functions like pathogenesis and immune evasion. Staphylococcus aureus is both a commensal of humans and a highly dangerous bacterial pathogen. EV production by S. aureus has been demonstrated and EVs shown to be cytolytic and play roles in immune activation. Studies of S. aureus EV composition have focused on their protein content and to date there are no reports describing the nucleic acid content of S. aureus EVs. S. aureus DNA and RNA are known to elicit an immune response however the mechanism(s) through which these nucleic acids gain access to the host cell is unknown. We hypothesize that EVs represent one mechanism through which S. aureus DNA/RNA is introduced into host cells. In this exploratory proposal we present preliminary data showing that RNA is located inside and associated with S. aureus EVs. We propose to identify the EV-associated RNA in EVs from five different S. aureus strains by RNA-sequencing, and to use that information to define the S. aureus EV-RNome – i.e. the RNA molecules found in EVs from all S. aureus strains. Defining the EV RNome will allow us to identify candidate RNA molecules for future analysis to determine their contribution to the immune stimulating properties of EVs.

项目概要/摘要 细胞外囊泡 (EV) 是由所有细胞产生和释放的纳米尺寸、膜封闭的囊泡 EV 包含多种生物分子，例如脂质、蛋白质和核酸。 参与广泛的功能，具体取决于祖细胞对革兰氏阳性细胞的 EV 的研究。 由于革兰氏阳性细胞壁的结构，细菌是一个相对较新的领域。 革兰氏阳性细菌不可能产生 EV，但最近的研究表明 EV 的产生。 多种革兰氏阳性细菌和涉及发病机制和免疫等重要功能的 EV 金黄色葡萄球菌既是人类的共生菌，也是高度危险的细菌病原体。 金黄色葡萄球菌可产生 EV，且 EV 具有溶细胞作用并在免疫中发挥作用 对金黄色葡萄球菌 EV 成分的研究主要集中在其蛋白质含量上，迄今为止还没有。 描述金黄色葡萄球菌 EV 的核酸含量的报告已知会引发 然而，免疫应答这些核酸进入宿主细胞的机制是 未知。我们认为 EV 代表了金黄色葡萄球菌 DNA/RNA 的引入机制。 在这个探索性提议中，我们提供了初步数据，表明 RNA 位于内部和 我们建议鉴定来自五种不同金黄色葡萄球菌 EV 的 EV 相关 RNA。 通过 RNA 测序分析金黄色葡萄球菌菌株，并使用该信息来定义金黄色葡萄球菌 EV-RNome，即 在所有金黄色葡萄球菌菌株的 EV 中发现的 RNA 分子定义 EV RNome 将使我们能够识别候选者。 用于未来分析的 RNA 分子，以确定它们对 EV 免疫刺激特性的贡献。