Differential microRNA expression between decidual and peripheral blood natural killer cells in early pregnancy
Differential microRNA expression between decidual and peripheral blood natural killer cells in early pregnancy
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DOI：
10.1093/humrep/dey323
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发表时间：
2018-12-01
影响因子：
6.1
通讯作者：
Gismondi, A.
中科院分区：
文献类型：
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作者：
Carlino, C.;Rippo, M. R.;Gismondi, A.
研究问题：与妊娠早期健康孕妇外周血（pb）中循环的NK细胞相比，蜕膜自然杀伤细胞（dNK）是否具有不同的microRNA （miRNA或miR）表达模式？答案：dNK细胞具有独特的miRNA谱，显示出一组miRNA的独家表达，并且与pbNK细胞共享的大多数miRNA显著上调或下调。已知情况：dNK细胞与pbNK细胞在表型和功能上都不同，它们的起源仍有争议。许多研究表明，mirna调节NK细胞生物学的几个重要方面，如发育、激活和效应功能。研究设计、大小、持续时间：从妊娠前三个月自愿终止妊娠的妇女（n = 7）收集基底蜕膜和外周血标本。然后通过细胞分选高度纯化dNK和pbNK细胞。参与者/材料，设置，方法：使用从新鲜分离和高度纯化的pbNK和dNK细胞中纯化的RNA，通过基于定量RT-PCR （qRT-PCR）的阵列分析miRNAs的表达。通过qRT-PCR检测验证阵列结果。应用生物信息学工具独创性途径分析（IPA）来确定经过验证的mirna靶向的细胞网络和相关的生物学功能。主要结果和机会的作用：在这里，我们从孕妇外周血和子宫蜕膜分离的NK细胞中鉴定出了表达差异最大的mirna。我们发现36个mirna仅在dNK细胞中表达，2个mirna仅在pbNK细胞中表达。此外，48种mirna在两种NK细胞制剂中均有表达，尽管表达水平不同：与pbNK细胞相比，dNK细胞中有28种表达上调，而15种表达下调。对选定的一组mirna （n = 11）的验证证实了9种mirna的差异表达：miR-10b和miR-214仅在dNK细胞中表达，miR-200a-3p仅在pbNK细胞中表达；与pbNK细胞相比，dNK细胞中miR-130b-3p、miR-125a-5p、miR-212-3p和miR-454表达上调，miR-210-3p和miR-132表达下调。IPA网络分析确定了一个连接所有mirna的单一网络，以及它们在几类功能中的重要作用：“机体损伤、生殖系统疾病、炎症性疾病”和“细胞发育”。这些miRNAs靶向的分子如argonaute 2、肿瘤蛋白p53、胰岛素等基因属于同一网络，并显著影响细胞分化和妊娠。局限性，谨慎原因：在本研究中，通过IPA鉴定了dNK和pbNK细胞中差异表达的mirna调节的细胞网络和生物学功能，仅考虑白细胞中“实验观察”到的分子和关系。这里分析的蜕膜细胞和pbNK细胞是一个异质群体，进一步的研究将有助于解开不同NK细胞亚群在miRNA产生方面是否存在差异。研究结果的更广泛意义：这是第一个描述dNK细胞与匹配的pbNK细胞在妊娠早期的不同miRNA表达谱的研究。我们的发现改善了dNK细胞生物学的知识体系，并强烈建议进一步研究人类dNK与pbNK细胞中差异表达的mirna的作用。我们的研究结果表明，特定的miRNA可以调节dNK细胞的起源和功能，突出了这种miRNA在人类发育和疾病中的潜在作用。研究经费/竞争利益(S)：这项工作得到了巴斯德研究所、博洛涅蒂科学基金会、FP6内的欧洲NoE EMBIC（合同编号LSHN-CT-2004-512040）、意大利理工学院、意大利国立大学、意大利国立大学部长、意大利国立大学（Ricerche Universitarie）和意大利国立理工大学的资助。没有需要申报的利益冲突。
STUDY QUESTION: Have decidual natural killer (dNK) cells a different microRNA (miRNA or miR) expression pattern compared to NK cells circulating in the peripheral blood (pb) of healthy pregnant women in the first trimester of gestation?SUMMARY ANSWER: dNK cells have a unique miRNA profile, showing exclusive expression of a set of miRNAs and significant up-or down-regulation of most of the miRNAs shared with pbNK cells.WHAT IS KNOWN ALREADY: dNK cells differ from pbNK cells both phenotypically and functionally, and their origin is still debated. Many studies have indicated that miRNAs regulate several important aspects of NK cell biology, such as development, activation and effector functions.STUDY DESIGN, SIZE, DURATION: Decidua basalis and peripheral blood specimens were collected from women (n = 7) undergoing voluntary termination of gestation in the first trimester of pregnancy. dNK and pbNK cells were then highly purified by cell sorting.PARTICIPANTS/MATERIALS, SETTING, METHODS: miRNAs expression was analysed by quantitative RT-PCR (qRT-PCR)-based arrays using RNA purified from freshly isolated and highly purified pbNK and dNK cells. Results from arrays were validated by qRT-PCR assays. The bioinformatics tool ingenuity pathway analysis (IPA) was applied to determine the cellular network targeted by validated miRNAs and the correlated biological functions.MAIN RESULTS AND THE ROLE OF CHANCE: Herein, we identified the most differentially expressed miRNAs in NK cells isolated from peripheral blood and uterine decidua of pregnant women. We found that 36 miRNAs were expressed only in dNK cells and two miRNAs only in pbNK cells. Moreover, 48 miRNAs were commonly expressed by both NK cell preparations although at different levels: 28 were upregulated in dNK cells, while 15 were downregulated compared to pbNK cells. Validation of a selected set (n = 11) of these miRNAs confirmed the differential expression of nine miRNAs: miR-10b and miR-214 expressed only in dNK cells and miR-200a-3p expressed only in pbNK cells; miR-130b-3p, miR-125a-5p, miR-212-3p and miR-454 were upregulated while miR-210-3p and miR-132 were downregulated in dNK cells compared to pbNK cells. IPA network analysis identified a single network connecting all the miRNAs as well as their significantinvolvement in several classes of functions: 'Organismal injury, Reproductive system disease, Inflammatory disease' and 'Cellular development'. These miRNAs target molecules such as argonaute 2, tumour protein p53, insulin and other genes that belong to the same network and significantly influence cell differentiation and pregnancy.LIMITATIONS, REASONS FOR CAUTION: In the present study, the cellular network and biological functions modulated by miRNAs differentially expressed in dNK and pbNK cells were identified by IPA considering only molecules and relationships that were with confidence 'experimentally observed' in leucocytes. The decidual and pbNK cells that were analysed here are a heterogeneous population and further study will help to disentangle whether there are differences in miRNA production by the different subsets of NK cells.WIDER IMPLICATIONS OF THE FINDINGS: This is the first study describing a different miRNA expression profile in dNK cells compared to matched pbNK cells during the first trimester of pregnancy. Our findings improved the body of knowledge on dNK cell biology and strongly suggest further investigation into the roles of miRNAs that are differentially expressed in human dNK compared to pbNK cells. Our results suggest that specific miRNAs can modulate dNK cell origin and functions, highlighting a potential role of this miRNA signature in human development and diseases.STUDY FUNDING/COMPETING INTEREST(S): This work was supported by grants from the Istituto Pasteur, Fondazione Cenci Bolognetti, the European NoE EMBIC within FP6 (Contract number LSHN-CT-2004-512040), Istituto Italiano di Tecnologia, and Ministero dell'Istruzione, dell'Universita e della Ricerca (Ricerche Universitarie), and from Universita Politecnica delle Marche. There are no conflicts of interest to declare.