Dix Andreas, Czakai Kristin, Leonhardt Ines, Schäferhoff Karin, Bonin Michael, Guthke Reinhard, Einsele Hermann, Kurzai Oliver, Löffler Jürgen, Linde Jörg
Systems Biology/Bioinformatics, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute Jena, Germany.
Department of Internal Medicine II, University Hospital of Würzburg Würzburg, Germany.
Front Microbiol. 2017 Feb 23;8:270. doi: 10.3389/fmicb.2017.00270. eCollection 2017.
Within the last two decades, the incidence of invasive fungal infections has been significantly increased. They are characterized by high mortality rates and are often caused by and . The increasing number of infections underlines the necessity for additional anti-fungal therapies, which require extended knowledge of gene regulations during fungal infection. MicroRNAs are regulators of important cellular processes, including the immune response. By analyzing their regulation and impact on target genes, novel therapeutic and diagnostic approaches may be developed. Here, we examine the role of microRNAs in human dendritic cells during fungal infection. Dendritic cells represent the bridge between the innate and the adaptive immune systems. Therefore, analysis of gene regulation of dendritic cells is of particular significance. By applying next-generation sequencing of small RNAs, we quantify microRNA expression in monocyte-derived dendritic cells after 6 and 12 h of infection with and as well as treatment with lipopolysaccharides (LPS). We identified 26 microRNAs that are differentially regulated after infection by the fungi or LPS. Three and five of them are specific for fungal infections after 6 and 12 h, respectively. We further validated interactions of miR-132-5p and miR-212-5p with immunological relevant target genes, such as , and , on both RNA and protein level. Our results indicate that these microRNAs fine-tune the expression of immune-related target genes during fungal infection. Beyond that, we identified previously undiscovered microRNAs. We validated three novel microRNAs via qRT-PCR. A comparison with known microRNAs revealed possible relations with the miR-378 family and miR-1260a/b for two of them, while the third one features a unique sequence with no resemblance to known microRNAs. In summary, this study analyzes the effect of known microRNAs in dendritic cells during fungal infections and proposes novel microRNAs that could be experimentally verified.
在过去二十年中，侵袭性真菌感染的发病率显著上升。其特点是死亡率高，通常由[具体真菌名称1]和[具体真菌名称2]引起。感染数量的增加凸显了额外抗真菌治疗的必要性，这需要深入了解真菌感染期间的基因调控。微小RNA是重要细胞过程的调节因子，包括免疫反应。通过分析它们对靶基因的调控和影响，可能开发出新的治疗和诊断方法。在这里，我们研究了真菌感染期间微小RNA在人树突状细胞中的作用。树突状细胞代表先天免疫系统和适应性免疫系统之间的桥梁。因此，分析树突状细胞的基因调控具有特别重要的意义。通过应用小RNA的下一代测序技术，我们对单核细胞衍生的树突状细胞在感染[具体真菌名称1]和[具体真菌名称2]6小时和12小时后以及用脂多糖（LPS）处理后的微小RNA表达进行了定量。我们鉴定出26种微小RNA在真菌或LPS感染后受到差异调节。其中分别有3种和5种在6小时和12小时后对真菌感染具有特异性。我们进一步在RNA和蛋白质水平上验证了miR - 132 - 5p和miR - 212 - 5p与免疫相关靶基因（如[具体靶基因1]、[具体靶基因2]和[具体靶基因3]）的相互作用。我们的结果表明，这些微小RNA在真菌感染期间微调免疫相关靶基因的表达。除此之外，我们还鉴定出了以前未发现的微小RNA。我们通过qRT - PCR验证了三种新的微小RNA。与已知微小RNA的比较显示，其中两种与miR - 378家族和miR - 1260a/b可能存在关系，而第三种具有独特的序列，与已知微小RNA没有相似之处。总之，本研究分析了真菌感染期间已知微小RNA在树突状细胞中的作用，并提出了可以通过实验验证的新微小RNA。
