Integrated transcriptomics and metabolomics approach to define pathogenicity of P.gingivalis

综合转录组学和代谢组学方法确定牙龈卟啉单胞菌的致病性

• 批准号: 10112887
• 负责人: Sasanka S Chukkapalli
• 金额: $ 0.89万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2021-06-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112887
• 关键词: Anaerobic Bacteria; Animal Model; Anti-Inflammatory Agents; Atherosclerosis; Bacterial Genes; Biological Markers; Cells; Chronic; Coronary artery; Coupled; Data; Development; Endothelial Cells; Endothelium; Environment; Event; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Gingiva; Goals; High Pressure Liquid Chromatography; Host Defense; Human; Immune response; Impairment; Infection; Infection Control; Inflammatory; Invaded; Lipoproteins; Messenger RNA; Metabolic; Metabolic Pathway; Molecular; Nature; Operon; Organism; Outcome; Pathogenesis; Pathogenicity; Pattern; Periodontal Diseases; Periodontitis; Porphyromonas gingivalis; Protein-arginine deiminase; Regulation; Resolution; Risk; Role; Signal Transduction; System; Systemic disease; Technology; Testing; Tooth Loss; Virulence; Virulence Factors; Work; chronic infection; design; effective therapy; endopeptidase Clp; endothelial dysfunction; gingipain; innovation; insight; lipidome; metabolome; metabolomics; microbial; mutant; next generation; pathogen; prevent; protein-histidine kinase; response; specific biomarkers; targeted treatment; transcriptome sequencing; transcriptomics; treatment strategy

Project abstract Porphyromonas gingivalis (Pg), a gram-negative asaccharolytic anaerobe, is a major causative pathogen of chronic periodontitis. Pg not only causes tooth loss, but is also associated with increased risk for other systemic diseases such as atherosclerosis. Endothelial cells provide an interface for the systemic dissemination of Pg. Pg evades endothelial cell defenses and persist within endothelial cells, thereby allowing chronic infections like periodontal disease and systemic diseases to develop. Pg persistence can also leads to endothelial dysfunction resulting in impaired inflammatory signals and atherosclerosis. Hence, it is critical to understand the mechanisms of Pg infection of endothelial cells. Our central hypothesis is that Pg invades endothelial cells and evades host defenses by regulating its gene expression and metabolic requirements to survive within endothelial cells. To test this hypothesis, we propose to employ an innovative approach of integrating transcriptomics and metabolomics to better understand the pathogenic nature of Pg and its interaction with the endothelial cells. This comprehensive approach of combining dual RNA-seq and metabolomics data will identify significant gene- metabolite integrated networks that are unique to Pg infection of endothelial cells. Consequently, these studies will define those metabolic changes that occur during intracellular adaptation and survival of Pg and provide much needed fundamental insights into periodontal disease and systemic diseases associated with Pg. In order to achieve this goal, we propose the following specific aims. Aim 1: To determine and compare the RNA expression patterns of P. gingivalis (W83, W83Δ0717 & W83Δ0717::0717) and host cells (HMVEC and HCAEC) during the invasion and persistence states. Aim 2: To define the metabolomic changes that occur during the invasion and persistence states of P. gingivalis (W83, W83Δ0717 & W83Δ0717::0717) in HMVEC and HCAEC cells. These studies will be the first to characterize the mRNA and metabolite patterns of both Pg and host cells during pathogenesis. The expected outcome of this work is a comprehensive understanding of what molecular mechanism(s) specific to gene-metabolite networks Pg utilizes during invasion and persistence states. The successful completion of the proposed studies will lay the groundwork for developing effective treatment strategies to prevent PD and its associated systemic diseases.

项目摘要 牙龈卟啉单胞菌（Porphyromonas gingivalis，Pg）是一种革兰氏阴性的溶砷厌氧菌，是牙龈炎的主要病原菌， 慢性牙周炎前列腺素不仅会导致牙齿脱落，而且还与其他全身性疾病的风险增加有关。 如动脉粥样硬化等疾病。内皮细胞为Pg的全身传播提供了界面。 逃避内皮细胞的防御，并在内皮细胞内持续存在，从而使慢性感染， 牙周病和全身性疾病的发展。PG持续存在也可导致内皮功能障碍 导致炎症信号受损和动脉粥样硬化。因此，了解 Pg感染内皮细胞的机制。我们的中心假设是Pg侵入内皮细胞， 通过调节其基因表达和代谢需求来逃避宿主防御，以在内皮细胞内存活。 细胞为了验证这一假设，我们建议采用一种创新的方法，将转录组学和 代谢组学，以更好地了解Pg的致病性及其与内皮细胞的相互作用。这 结合双重RNA-seq和代谢组学数据的综合方法将确定重要的基因， 代谢物整合网络是Pg感染内皮细胞所特有的。因此，这些研究 将定义在Pg的细胞内适应和存活期间发生的那些代谢变化，并提供 牙周病和与Pg相关的系统性疾病急需的基本见解。为了 为达致这个目标，我们提出以下具体目标。目的1：测定和比较 牙龈卟啉单胞菌（W83、W83Δ0717和W83Δ0717：：0717）和宿主细胞（HMVEC和HCAEC）的表达模式 在入侵和持续状态期间。目的2：确定在代谢过程中发生的代谢组学变化。 牙龈卟啉单胞菌（W83，W83Δ0717和W83Δ0717：：0717）在HMVEC和HCAEC中侵袭和持久状态 细胞这些研究将是第一个描述Pg和宿主细胞的mRNA和代谢物模式的研究 在发病过程中。这项工作的预期成果是全面了解什么分子 Pg在入侵和持续状态期间利用的基因代谢网络特有的机制。的 成功完成拟议的研究将为开发有效的治疗方法奠定基础 预防PD及其相关系统性疾病的策略。