Kalea A Z, Hoteit R, Suvan J, Lovering R C, Palmen J, Cooper J A, Khodiyar V K, Harrington Z, Humphries S E, D'Aiuto F
Centre for Cardiovascular Genetics, Institute of Cardiovascular Science, University College London, London, UK
Centre for Cardiovascular Genetics, Institute of Cardiovascular Science, University College London, London, UK.
J Dent Res. 2015 Mar;94(3 Suppl):59S-69S. doi: 10.1177/0022034514568197. Epub 2015 Jan 28.
Increased local immune and inflammatory responses in obese individuals with periodontitis may explain the aggressive clinical presentation and altered treatment response when compared to that of normal weight subjects. Our goal was to identify any differences in microRNA (miRNA) expression profiles of gingival tissue in periodontitis when obesity is present, which may suggest novel molecular pathways that this miRNA network may affect. Total RNA was extracted from gingival tissue biopsies collected from normal weight and obese individuals with periodontitis; miRNA expression profiling was performed with Affymetrix GeneChip miRNA 3.0 arrays; and results were validated with quantitative reverse transcription polymerase chain reaction (qRT-PCR). In silico identification of previously confirmed miRNA gene targets was conducted through miRTarBase and miRWalk databases, and pathway enrichment analysis identified enriched miRNA gene sets. Expression of selected genes in the same biopsy samples was tested with qRT-PCR. The gingival tissue miRNA profile of obese patients, compared to that of normal weight patients, showed 13 upregulated and 22 downregulated miRNAs, among which miR-200b was validated by qRT-PCR to be significantly increased in obesity. Functional analysis of 51 experimentally validated miR-200b gene targets identified enrichment of genes involved in cell motility, differentiation, DNA binding, response to stimulus, and vasculature development pathways not previously identified in the obesity-specific disease profile. Furthermore, the expression of the miR-200b gene targets ZEB1/2, GATA2, and KDR was confirmed by qRT-PCR as being lower in obese patients with periodontitis versus normal weight patients, suggesting a role of miR-200b in regulation of a set of gene targets and biological pathways relevant to wound healing and angiogenesis. Functional studies to explore the role of miR-200b in the above processes may offer new insights on putative therapeutic targets for this group of patients.
与正常体重受试者相比,肥胖牙周炎患者局部免疫和炎症反应增强,这可能解释了其侵袭性临床表现和治疗反应的改变。我们的目标是确定肥胖时牙周炎患者牙龈组织中微小RNA(miRNA)表达谱的差异,这可能提示该miRNA网络可能影响的新分子途径。从正常体重和肥胖牙周炎患者的牙龈组织活检样本中提取总RNA;使用Affymetrix GeneChip miRNA 3.0阵列进行miRNA表达谱分析;并通过定量逆转录聚合酶链反应(qRT-PCR)验证结果。通过miRTarBase和miRWalk数据库对先前已确认的miRNA基因靶点进行计算机识别,并通过通路富集分析确定富集的miRNA基因集。用qRT-PCR检测同一活检样本中选定基因的表达。与正常体重患者相比,肥胖患者的牙龈组织miRNA谱显示13种miRNA上调,22种miRNA下调,其中miR-200b经qRT-PCR验证在肥胖患者中显著增加。对51个经实验验证的miR-200b基因靶点进行功能分析,发现参与细胞运动、分化、DNA结合、对刺激的反应以及血管发育途径的基因富集,这些途径在肥胖特异性疾病谱中尚未被发现。此外,qRT-PCR证实肥胖牙周炎患者与正常体重患者相比,miR-200b基因靶点ZEB1/2、GATA2和KDR的表达较低,提示miR-200b在调控一组与伤口愈合和血管生成相关的基因靶点和生物学途径中发挥作用。探索miR-200b在上述过程中作用的功能研究可能为该组患者的潜在治疗靶点提供新的见解。