牙-肾综合征牙龈组织的转录组分析。
Transcriptome analysis of gingival tissues of enamel-renal syndrome.
机构信息
Department of Dentistry, School of Dentistry, National Taiwan University, Taipei City, Taiwan.
Graduate Institute of Clinical Dentistry, School of Dentistry, National Taiwan University, Taipei City, Taiwan.
出版信息
J Periodontal Res. 2019 Dec;54(6):653-661. doi: 10.1111/jre.12666. Epub 2019 May 27.
BACKGROUND AND OBJECTIVE
Biallelic loss-of-function mutations of human FAM20A have been known to cause enamel-renal syndrome (ERS), featured by agenesis of dental enamel, nephrocalcinosis, and other orodental abnormalities, including gingival hyperplasia. However, while the histopathology of this gingival anomaly has been analyzed, its underlying molecular mechanism remains largely unknown. This study aimed to unravel the pathogenesis of gingival hyperplasia in ERS.
METHODS
Whole-exome sequencing was conducted for an ERS case. Transcriptome analyses, using RNA sequencing, of the patient's gingiva were performed to unravel dysregulated molecules and aberrant biological processes underlying the gingival pathology of ERS, which was further confirmed by histology and immunohistochemistry.
RESULTS
Two novel frameshift FAM20A mutations in Exon 1 (g.5417delG; c.129delG; p.Cys44Alafs101) and Exon 5 (g.62248_62249delAG; c.734_735delAG; p.Glu245Glyfs11) were identified. Transcriptional profiling of patient's gingival tissue revealed a total of 1683 genes whose expression had increased (1129 genes) or decreased (554 genes) at least 2-fold compared to control gingival tissues. There were 951 gene ontology (GO) terms of biological process being significantly over-represented or under-represented. While GOs involved in extracellular matrix organization, angiogenesis, biomineralization, and epithelial cell proliferation appeared to be activated in ERS gingiva, genes related to keratinocyte differentiation, epithelial development, and keratinization were of decreased expression. FAM20A immunohistochemistry revealed a strong reactivity at the suprabasal layers of epithelium in control gingiva but showed a significantly diminished and scattered signal in ERS tissues. For genes showing significant over-expression in the transcriptome analyses, namely ALPL, SPARC, and ACTA2, an increased immunoreactivity was observed.
CONCLUSION
Our results unraveled a potential role for FAM20A in homeostasis of both gingival epithelium and connective tissues.
背景与目的
人类 FAM20A 的双等位基因功能丧失突变已被证实可导致牙釉质-肾综合征（ERS），其特征为牙釉质发育不全、肾钙质沉着症和其他口颌异常，包括牙龈增生。然而，尽管已经对这种牙龈异常的组织病理学进行了分析，但它的潜在分子机制在很大程度上仍然未知。本研究旨在揭示 ERS 中牙龈增生的发病机制。
方法
对 ERS 病例进行全外显子组测序。使用 RNA 测序对患者的牙龈进行转录组分析，以揭示 ERS 牙龈病理学背后失调的分子和异常的生物学过程，这些过程进一步通过组织学和免疫组织化学得到证实。
结果
在 Exon 1（g.5417delG；c.129delG；p.Cys44Alafs101）和 Exon 5（g.62248_62249delAG；c.734_735delAG；p.Glu245Glyfs11）中发现了两个新的移码 FAM20A 突变。与对照牙龈组织相比，患者牙龈组织的转录谱显示总共有 1683 个基因的表达增加（1129 个基因）或减少（554 个基因）至少 2 倍。有 951 个基因本体（GO）术语的生物学过程显著过表达或低表达。虽然在 ERS 牙龈中似乎激活了与细胞外基质组织、血管生成、生物矿化和上皮细胞增殖相关的 GO，但与角蛋白细胞分化、上皮发育和角化相关的基因表达减少。FAM20A 免疫组织化学显示在对照牙龈的上皮基底层有强烈的反应性，但在 ERS 组织中反应性明显减弱和分散。对于转录组分析中显示出显著过表达的基因，即 ALPL、SPARC 和 ACTA2，观察到免疫反应性增加。
结论
我们的研究结果揭示了 FAM20A 在牙龈上皮和结缔组织的稳态中可能发挥作用。
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