Diabetes-enhanced Experimental Periodontitis

翻译后摘要：糖尿病增加了细菌感染的风险和频率，并改变了适应性免疫 反应令人惊讶的是，关于糖尿病对树突细胞的影响以及它如何增加， 牙周炎的易感性。直到最近，由于技术有限， 定义树突状细胞的特定行为以及在活动期间与其他细胞类型相互作用的能力 牙周病单细胞转录组学的新进展，如单细胞RNA-seq（scRNA-seq）， 提供了一个指数增加，我们的能力，解剖行为的特定细胞类型在各种疾病 条件这些研究将阐明糖尿病如何改变树突细胞的功能， 会导致牙周炎已添加新的Prel数据，从而导致完全修订的提案 这表明糖尿病改变了体内树突状细胞基因表达，如通过scRNA-seq评估的。研究 建议涉及最近获得的scRNA-seq数据的复杂生物信息学分析和一种新的 实验方法，空间分辨转录组学，提供新的见解，糖尿病如何可能影响 树突状细胞活性。目标1中该方法的风险通过已识别潜在风险的Prel数据缓解 牙周病活动期树突状细胞中牙周组织上调或下调的关键基因 进展，这将通过scRNA-seq结果的生物信息学分析进一步研究。此外，本发明还提供了一种方法， 在目标1中提出了转化研究作为治疗干预的临床前步骤。功能研究 在目标2中提出，在体外检查候选基因，以建立 候选基因和功能结果。Aim 3中的实验将使用空间转录组学来定义 糖尿病可能会改变DC和其他白细胞的空间区室， 牙周炎和调查共定位的细胞和炎症基因表达模式，以更好地 了解他们的互动。总而言之，这个提议结合了一种无偏见的假设方法 通过功能性体外和体内方法进行假设检验和建立机制。

Abstract: Diabetes increases the risk and frequency of bacterial infection and alters the adaptive immune response. Surprisingly little is known about the impact of diabetes on dendritic cells and how it may increase susceptibility to periodontitis. The lack of understanding until recently has been hampered by limited technical ability to define the specific behavior of dendritic cells and interactions with other cell types during active periodontal disease. New advances in single cell transcriptomics such as single cell RNA-seq (scRNA-seq) have provided an exponential increase in our ability to dissect the behavior of specific cell types under various disease conditions. The studies proposed will elucidate how diabetes alters dendritic cell function in a way that contributes to periodontitis. New Prel Data has been added resulting a completely revised proposal demonstrating that diabetes alters dendritic cell gene expression in vivo as assessed by scRNA-seq. The studies proposed involve sophisticated bioinformatic analysis of scRNA-seq data very recently obtained and a new experimental approach, spatially resolved transcriptomics, to provide new insight on how diabetes may affect dendritic cell activity. The risk in this approach in Aim 1 is mitigated by Prel Data that have identified potential key genes that are up- or down-regulated in the periodontium in dendritic cells during active periodontal disease progression, which will be further investigated by bioinformatic analysis of scRNA-seq results. In addition, translational studies are proposed in Aim 1 as a preclinical step for therapeutic intervention. Functional studies are proposed in Aim 2 to examine gene candidates in vitro to establish cause and effect relationships between candidate genes and functional outcomes. Experiments in Aim 3 will use spatial transcriptomics to define how diabetes may alter the spatial compartments in which DC and other leukocytes are found during the initiation of periodontitis and investigate co-localization of cells and pattern of inflammatory gene expression to better understand their interaction. Taken together this proposal combines an unbiased approach for hypothesis discovery with functional in vitro and in vivo methods for hypothesis testing and establishing mechanisms.