miR-34a as hub of T cell regulation networks
miR-34a as hub of T cell regulation networks
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DOI：
10.1186/s40425-019-0670-5
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发表时间：
2019-07-16
影响因子：
10.9
通讯作者：
Meese, Eckart
中科院分区：
文献类型：
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作者：
Hart, Martin;Walch-Rueckheim, Barbara;Meese, Eckart
背景：微（mi） rna越来越被认为是免疫细胞功能的中枢调节因子。虽然已经预测mirna具有多个靶标，但这些预测中的大多数仍有待实验证实。在这里，我们分析了miR-34a，一种众所周知的肿瘤抑制因子，其靶向参与白细胞免疫系统过程的基因。方法：采用芯片方法，将miRNA靶标预测与多组学富集分析网络工具GeneTrail2相结合，鉴定参与基因本体免疫系统过程亚类的miR-34a靶基因。结果：在该亚类的193个预测靶基因中，我们实验检测了22个靶基因，并通过双荧光素酶测定证实了miR-34a与14个靶基因的结合，包括VAMP2、IKBKE、MYH9、MARCH8、KLRK1、CD11A、TRAFD1、CCR1、PYDC1、PRF1、PIK3R2、PIK3CD、AP1B1和ADAM10。通过用miR-34a转染Jurkat、原代CD4（+）和CD8(+) T细胞，我们证明miR-34a的异位表达导致内源性VAMP2和CD11A水平降低，这是所分析亚类的核心。活化的CD8(+) T细胞中miR-34a过表达的功能下游分析显示PRF1分泌明显减少。结论：通过同时靶向14种mrna， miR-34a作为T细胞调节网络的主要枢纽，表明在广泛的肿瘤背景下，利用miR-34a作为干预靶点来调节T细胞的免疫反应性。
Background: Micro(mi)RNAs are increasingly recognized as central regulators of immune cell function. While it has been predicted that miRNAs have multiple targets, the majority of these predictions still await experimental confirmation. Here, miR-34a, a well-known tumor suppressor, is analyzed for targeting genes involved in immune system processes of leucocytes.Methods: Using an in-silico approach, we combined miRNA target prediction with GeneTrail2, a web tool for Multi-omics enrichment analysis, to identify miR-34a target genes, which are involved in the immune system process subcategory of Gene Ontology.Results: Out of the 193 predicted target genes in this subcategory we experimentally tested 22 target genes and confirmed binding of miR-34a to 14 target genes including VAMP2, IKBKE, MYH9, MARCH8, KLRK1, CD11A, TRAFD1, CCR1, PYDC1, PRF1, PIK3R2, PIK3CD, AP1B1, and ADAM10 by dual luciferase assays. By transfecting Jurkat, primary CD4(+) and CD8(+) T cells with miR-34a, we demonstrated that ectopic expression of miR-34a leads to reduced levels of endogenous VAMP2 and CD11A, which are central to the analyzed subcategories. Functional downstream analysis of miR-34a over-expression in activated CD8(+) T cells exhibits a distinct decrease of PRF1 secretion.Conclusions: By simultaneous targeting of 14 mRNAs miR-34a acts as major hub of T cell regulatory networks suggesting to utilize miR-34a as target of intervention towards a modulation of the immune responsiveness of T-cells in a broad tumor context.