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（Pg）是一种革兰氏阴性解糖厌氧菌，是牙龈卟啉单胞菌的主要致病菌。 慢性牙周炎不仅会导致牙齿脱落，还会增加其他系统性风险。 动脉粥样硬化等疾病为 Pg 的全身传播提供了界面。 逃避内皮细胞防御并在内皮细胞内持续存在，从而导致慢性感染，例如 牙周病和全身性疾病的发展也可能导致内皮功能障碍。 导致炎症信号受损和动脉粥样硬化。因此，了解这一点至关重要。 Pg 感染内皮细胞的机制 我们的中心假设是 Pg 侵入内皮细胞并 通过调节其基因表达和代谢需求来逃避宿主防御，以在内皮细胞中生存 为了检验这一假设，我们建议采用一种整合转录组学和细胞学的创新方法。 代谢组学，以更好地了解 Pg 的致病性质及其与内皮细胞的相互作用。 结合双 RNA 测序和代谢组学数据的综合方法将识别重要基因 这些研究测试了内皮细胞 Pg 感染所特有的代谢整合网络。 将定义 Pg 的细胞内适应和生存过程中发生的代谢变化，并提供 迫切需要对与 Pg 相关的牙周病和全身性疾病的基本见解。 为了实现这一目标，我们提出以下具体目标 1：确定和比较 RNA。 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 目标 2：确定侵袭和持续状态期间发生的代谢组变化。 HMVEC 和 HCAEC 中牙龈卟啉单胞菌 (W83、W83Δ0717 和 W83Δ0717::0717) 的入侵和持续状态 这些研究将首次描述 Pg 和宿主细胞的 mRNA 和代谢模式。 这项工作的预期结果是全面了解什么是分子。 Pg 在入侵和持久状态期间利用基因代谢网络特有的机制。 成功完成拟议研究将为开发有效治疗奠定基础 预防帕金森病及其相关全身性疾病的策略。