Deep sequencing of small RNAs from human skin reveals major alterations in the psoriasis miRNAome
Deep sequencing of small RNAs from human skin reveals major alterations in the psoriasis miRNAome
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DOI：
10.1093/hmg/ddr331
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发表时间：
2011-10-15
影响因子：
3.5
通讯作者：
Bowcock, Anne M.
中科院分区：
文献类型：
--
作者：
Joyce, Cailin E.;Zhou, Xiang;Bowcock, Anne M.
银屑病是一种慢性和复杂的炎症性皮肤病，病变表现出显著改变的mRNA表达谱。然而，我们对小rna的表达知之甚少。在这里，我们描述了在一个大型患者队列中使用下一代测序对正常和银屑病皮肤miRNAome的综合分析。我们生成了6.7 x 10(8)个小RNA读取，代表了717个已知的和284个假定的新型microRNAs （miRNAs）。我们还观察到，在正常和银屑病皮肤中，isomir和miRNA*s广泛表达，源自已知和新的miRNA位点，miRNA编辑的频率较低。采用qRT-PCR技术，在皮肤和其他人体组织或细胞系中证实了新mirna的表达和加工。80个已知和18个新的mirna在银屑病皮肤中差异表达2-42倍。特别重要的是，来自miR-203位点反义链的一种经过验证的新型miRNA的2.7倍上调，该miRNA在上皮分化中起作用。其他差异表达的miRNA包括造血特异性miRNA，如miR-142-3p和miR-223/223*，以及血管生成性miRNA，如miR-21、miR-378、miR-100和miR-31，这是银屑病皮肤中表达最高的miRNA。这些mirna的功能与银屑病病变的炎症和增生性表型一致。对差异表达的miRNA进行原位杂交发现，在银屑病病变中，角质形成细胞来源的miRNA miR-135b和造血特异性miRNA miR-142-3p的表皮浸润呈层状表达。该研究为健康和患病皮肤中mirna的功能表征奠定了关键框架。
Psoriasis is a chronic and complex inflammatory skin disease with lesions displaying dramatically altered mRNA expression profiles. However, much less is known about the expression of small RNAs. Here, we describe a comprehensive analysis of the normal and psoriatic skin miRNAome with next-generation sequencing in a large patient cohort. We generated 6.7 x 10(8) small RNA reads representing 717 known and 284 putative novel microRNAs (miRNAs). We also observed widespread expression of isomiRs and miRNA*s derived from known and novel miRNA loci, and a low frequency of miRNA editing in normal and psoriatic skin. The expression and processing of selected novel miRNAs were confirmed with qRT-PCR in skin and other human tissues or cell lines. Eighty known and 18 novel miRNAs were 2-42-fold differentially expressed in psoriatic skin. Of particular significance was the 2.7-fold upregulation of a validated novel miRNA derived from the antisense strand of the miR-203 locus, which plays a role in epithelial differentiation. Other differentially expressed miRNAs included hematopoietic-specific miRNAs such as miR-142-3p and miR-223/223*, and angiogenic miRNAs such as miR-21, miR-378, miR-100 and miR-31, which was the most highly upregulated miRNA in psoriatic skin. The functions of these miRNAs are consistent with the inflammatory and hyperproliferative phenotype of psoriatic lesions. In situ hybridization of differentially expressed miRNAs revealed stratified epidermal expression of an uncharacterized keratinocyte-derived miRNA, miR-135b, as well as the epidermal infiltration of the hematopoietic-specific miRNA, miR-142-3p, in psoriatic lesions. This study lays a critical framework for functional characterization of miRNAs in healthy and diseased skin.