Laser capture microdissection followed by next-generation sequencing identifies disease-related microRNAs in psoriatic skin that reflect systemic microRNA changes in psoriasis
Laser capture microdissection followed by next-generation sequencing identifies disease-related microRNAs in psoriatic skin that reflect systemic microRNA changes in psoriasis
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DOI：
10.1111/exd.12604
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发表时间：
2015-03-01
影响因子：
3.6
通讯作者：
Skov, Lone
中科院分区：
文献类型：
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作者：
Lovendorf, Marianne B.;Mitsui, Hiroshi;Skov, Lone
牛皮癣是一种以皮肤为表现的全身性疾病。MicroRNAs(MiRNAs)是一种在银屑病皮肤中差异表达的非编码小RNA分子；然而，在银屑病皮损中只发现了少数细胞和区域特异性的miRNAs。我们用激光捕获显微切割(LCM)和下一代测序(NGS)技术研究了6例银屑病患者皮肤表皮(Epi)和真皮炎性浸润物(RD)中miRNA的表达谱。与正常银屑病皮肤相比，我们在银屑病斑块中发现了24个去调控的miRNAs，在RD中发现了37个去调控的miRNAs(FCH>；2，FDR<；0.05)。有趣的是，最近发现银屑病患者外周血单个核细胞(PBMC)中的9个miRNAs，包括miR-193B和miR-223，在银屑病患者的外周血单个核细胞(PBMC)中处于失控状态。应用流式细胞仪和qRT-PCR检测发现，miR-193B和miR-223在Th17细胞中均有表达。总之，我们证明了LCM和NGS相结合提供了一种强有力的方法来探索银屑病患者皮肤表皮和真皮间隔中miRNA的全球表达。此外，我们的结果表明，RD中局部miRNA的变化反映在循环免疫细胞中，提示miRNAs可能参与了银屑病的发病。
Psoriasis is a systemic disease with cutaneous manifestations. MicroRNAs (miRNAs) are small non-coding RNA molecules that are differentially expressed in psoriatic skin; however, only few cell-and region-specific miRNAs have been identified in psoriatic lesions. We used laser capture microdissection (LCM) and next-generation sequencing (NGS) to study the specific miRNA expression profiles in the epidermis (Epi) and dermal inflammatory infiltrates (RD) of psoriatic skin (N = 6). We identified 24 deregulated miRNAs in the Epi and 37 deregulated miRNAs in the RD of psoriatic plaque compared with normal psoriatic skin (FCH > 2, FDR < 0.05). Interestingly, 9 of the 37 miRNAs in RD, including miR-193b and miR-223, were recently described as deregulated in circulating peripheral blood mononuclear cells (PBMCs) from patients with psoriasis. Using flow cytometry and qRT-PCR, we found that miR-193b and miR-223 were expressed in Th17 cells. In conclusion, we demonstrate that LCM combined with NGS provides a robust approach to explore the global miRNA expression in the epidermal and dermal compartments of psoriatic skin. Furthermore, our results indicate that the altered local miRNA changes seen in the RD are reflected in the circulating immune cells, suggesting that miRNAs may contribute to the pathogenesis of psoriasis.