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

项目摘要 牙龈卟啉单胞菌(Pg)是一种革兰氏阴性厌氧菌，是引起牙周炎的主要病原菌。 慢性牙周炎。前列腺癌不仅会导致牙齿脱落，还会增加其他系统性疾病的风险。 动脉粥样硬化等疾病。内皮细胞为PG的全身性传播提供了接口。皮格 逃避内皮细胞防御并持续存在于内皮细胞内，从而允许像这样的慢性感染 牙周病和全身性疾病的发展。PG持续存在也会导致内皮功能障碍 导致炎症信号受损和动脉粥样硬化。因此，理解 PG感染内皮细胞的机制。我们的中心假设是PG侵入内皮细胞并 通过调节其基因表达和代谢要求来逃避宿主防御，以在内皮细胞内生存 细胞。为了验证这一假设，我们建议采用一种创新的方法，将转录和 代谢组学，以更好地了解PG的致病本质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识别重要的基因。 PG感染内皮细胞所特有的代谢产物整合网络。因此，这些研究 将定义在PG细胞内适应和生存过程中发生的代谢变化，并提供 急需对牙周病和与牙周炎相关的全身疾病的基本见解。按顺序 为实现这一目标，我们提出了以下具体目标。目的1：测定和比较核糖核酸 牙龈假单胞菌(W83，W83Δ0717和W83Δ0717：：0717)和宿主细胞(HMVEC和HCAEC)的表达模式 在入侵和持久状态期间。目标2：定义在代谢过程中发生的代谢变化 牙周炎假单胞菌(W83，W83Δ0717和W83Δ0717：：0717)在人脐静脉内皮细胞中的侵袭和持续状态 细胞。这些研究将首次对PG和宿主细胞的mRNA和代谢产物模式进行表征 在发病过程中。这项工作的预期结果是全面了解什么分子 PG在入侵和持续状态中利用的基因-代谢网络特有的机制(S)。这个 拟议研究的成功完成将为开发有效的治疗方法奠定基础 预防帕金森病及其相关系统性疾病的策略。

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.