MicroRNA expression in gingival tissues in periodontal health and disease

描述（由申请人提供）：MicroRNA （miRNA）控制最近成为基因表达和蛋白质合成的关键调节因子，影响体内平衡、健康和疾病。目前还没有关于不同牙周健康和疾病状态下牙龈组织中mirna的差异、表达这些mirna的细胞亚群或这些短RNA序列的功能含义的信息。在NICDR支持的早期工作中，我们收集了慢性或侵袭性牙周炎患者的临床健康和病变牙龈组织标本，研究了牙龈组织转录组及其与牙周炎临床、微生物和血清学标志物的关系。在当前提案的第一阶段，我们打算利用上述研究中242个可用的存档组织标本，这些标本来自101个完全表型化的患者（53名慢性牙周炎患者和48名侵袭性牙周炎患者），以检查牙周健康和疾病状态以及不同类型牙周病变中牙龈组织中mirna的发生。接下来，我们将使用生物信息学方法来鉴定鉴定出的mirna优先结合的潜在基因靶点，并将在我们自己的全基因组转录组数据库中验证这些预测的靶点是否确实在牙龈中下调。随后，我们将从新招募的个体中收集牙龈组织，我们将确定在整个组织裂解物中发现的mirna差异表达的特定细胞群，并将确认它们的靶基因是否也在这些细胞类型中被抑制。最后，我们将在适当的细胞培养模型中使用一种新的miRNA抑制方法，开始对鉴定的miRNA序列进行功能表征。拟议的研究将以具有成本效益的方式在短时间内解决知识方面的重大差距，并将促进我们对牙周病病理生物学的理解。

DESCRIPTION (provided by applicant): MicroRNA (miRNA) control has recently emerged as a critical regulator of gene expression and protein synthesis, affecting homeostasis, health and disease. No information is available on the differential occurrence of miRNAs in the gingival tissues in various states of periodontal health and disease, on the cell sub- populations that express these miRNAs, or on the functional implications of these short RNA sequences. In our earlier work supported by NICDR, we have collected specimens from clinically healthy and diseased gingival tissues from patients with chronic or aggressive periodontitis, and have studied the gingival tissue transcriptome and its association with clinical, microbiological and serological markers of periodontitis. In the first phase of the current proposal, we intent to utilize 242 available archived tissue specimens from the above study, obtained from a subset of 101 thoroughly phenotyped patients (53 with chronic and 48 with aggressive periodontitis), to examine the occurrence of miRNAs in the gingival tissues in states of periodontal health and disease and in different types of periodontal lesions. Next, we will use bio-informatics approaches to identify potential gene targets to which the identified miRNAs bind preferentially, and will validate in our own full- genome transcriptomic database whether these predicted targets were indeed down-regulated in the gingiva. We will subsequently harvest gingival tissues from de novo recruited individuals, we will identify the specific cell populations that account for the miRNAs found in the previous step to be differentially expressed in whole tissue lysates, and will confirm whether their target genes are also repressed in these cell types. Lastly, we will use a novel miRNA inhibition approach in appropriate cell culture models to begin to functionally characterize the identified miRNA sequences. The proposed studies will address a significant gap in knowledge in a cost-effective manner and in a short period of time, and will advance our understanding of the pathobiology of periodontal diseases.