Identification of common lupus-disease related microRNA expression pattern in three different murine models of lupus

In this study, miRNA expression in splenic lymphocytes from three genetically disparate lupus-prone mouse models (MRL-lpr, B6-lpr and NZB/WF1) were profiled. 49 miRNAs were found to be differentially expressed in MRL-lpr mice compared to MRL mice; and 24 miRNAs were differentially expressed in B6-lpr mice compared to B6 mice. Among these dysregulated miRNAs, we noted that 15 miRNAs were common to both lpr strains. Interestingly, microarray analysis of NZB/W and NZW at 3 months of age, an age when overt lupus disease is not evident in NZB/W mice, revealed that only one miRNA, miR-148a was significantly upregulated in NZB/W mice. The aim of this porject is to determine the common miRNA expression changes in splenocytes from different strains of murine lupus models. The splenocytes were prepared from genetically lupus-prone female mice including MRL/MpJ-Faslpr/J (MRL-lpr), NZBWF1/J (NZB/W), B6.MRL-Faslpr/J (B6-lpr) and their control mice MRL/MpJ (MRL), NZW/LacJ (NZW) and C57BL/6J (B6) mice (The Jackson laboratory, ME). Total RNAs, containing miRNAs were isolated from whole splenocytes using mirVana miRNA isolation kits (Ambion) following manufacturer's instructions and sent to LC Sciences (http://www.lcsciences.com/) for the microarray assay. The mouse miRNA array chips (Chip ID miRMouse 12.0 version), which included 617 unique, mature, mouse miRNA, based on the Sanger miRBase Release 12.0, were used in the assay.本研究对三种遗传背景迥异的自发性狼疮小鼠模型（MRL-lpr、B6-lpr与NZB/WF1）的脾脏淋巴细胞微小RNA（microRNA，miRNA）表达谱进行了系统分析。相较于同品系对照小鼠MRL，MRL-lpr小鼠体内有49个miRNA呈现显著差异表达；相较于B6对照小鼠，B6-lpr小鼠体内则有24个miRNA呈现显著差异表达。在上述表达失调的miRNA中，本研究发现15个miRNA在两种lpr模型小鼠中均存在异常表达。 值得关注的是，对3月龄的NZB/W与NZW小鼠开展芯片分析时发现（该月龄下NZB/W小鼠尚未出现显性狼疮症状），仅miR-148a这一种miRNA在NZB/W小鼠体内呈现显著上调表达。本研究旨在明确不同品系自发性狼疮小鼠模型脾脏细胞中miRNA的共同表达变化规律。 本研究的样本取自自发性狼疮易感雌性小鼠，涵盖模型组小鼠MRL/MpJ-Faslpr/J（即MRL-lpr）、NZBWF1/J（即NZB/W）、B6.MRL-Faslpr/J（即B6-lpr），以及对应的对照组小鼠MRL/MpJ（MRL）、NZW/LacJ（NZW）与C57BL/6J（B6），所有小鼠均购自美国缅因州杰克逊实验室（The Jackson Laboratory, ME）。研究人员严格遵循制造商说明书，采用mirVana miRNA分离试剂盒（Ambion公司出品）从全脾脏细胞中提取包含miRNA在内的总RNA，并将样本送至LC Sciences公司（http://www.lcsciences.com/）进行芯片检测。本次实验使用的小鼠miRNA芯片（芯片编号miRMouse 12.0版）基于Sanger miRBase 12.0版数据库构建，可覆盖617种独特的成熟小鼠miRNA。