深度测序短 RNA 揭示干燥综合征患者小唾液腺中的新型 microRNAs。
Deep sequencing of short RNAs reveals novel microRNAs in minor salivary glands of patients with Sjögren's syndrome.
机构信息
Sjögren's Syndrome Clinic, Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, NIH, Bethesda, MD, USA.
出版信息
Oral Dis. 2012 Mar;18(2):127-31. doi: 10.1111/j.1601-0825.2011.01849.x. Epub 2011 Sep 6.
OBJECTIVES
Sjögren's syndrome is a complex autoimmune disease of the salivary gland with an unknown etiology, so a thorough characterization of the transcriptome would facilitate our understanding of the disease. We use ultradeep sequencing of small RNAs from patients with Sjögren's syndrome and healthy volunteers, primarily to identify and discover novel miRNA sequences that may play a role in the disease.
METHODS
Total RNA was isolated from minor salivary glands of healthy volunteers and patients with either high or low salivary flow and sequenced on the SOLiD platform. Prediction of mature miRNAs from the sequenced reads was carried out using miRanalyzer, and expression was validated using Taqman qPCR assays.
RESULTS
We validated the presence of six previously unidentified miRNA sequences in patient samples and in several cell lines. One of the validated novel miRNAs shows promise as a biomarker for salivary function.
CONCLUSION
Sequencing small RNAs in the salivary gland is largely unprecedented, but here, we show the feasibility of discovering novel miRNAs and disease biomarkers by sequencing the transcriptome.
目的
干燥综合征是一种病因不明的唾液腺自身免疫性疾病，因此对转录组进行全面描述将有助于我们了解该疾病。我们主要使用来自干燥综合征患者和健康志愿者的小 RNA 的超深度测序来识别和发现可能在疾病中发挥作用的新型 miRNA 序列。
方法
从小唾液腺中分离健康志愿者和唾液流量高或低的患者的总 RNA，并在 SOLiD 平台上进行测序。使用 miRanalyzer 从测序的读段中预测成熟的 miRNA，并使用 Taqman qPCR 检测进行表达验证。
结果
我们在患者样本和几种细胞系中验证了六种先前未鉴定的 miRNA 序列的存在。其中一种经过验证的新型 miRNA 有望成为唾液功能的生物标志物。
结论
对唾液腺中小 RNA 进行测序在很大程度上是前所未有的，但在这里，我们展示了通过对转录组进行测序发现新型 miRNA 和疾病生物标志物的可行性。
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