Regulation of Cathepsin E gene expression by the transcription factor Kaiso in MRL/lpr mice derived CD4+T cells
Regulation of Cathepsin E gene expression by the transcription factor Kaiso in MRL/lpr mice derived CD4+T cells
复制标题
DOI：
10.1038/s41598-019-38809-y
复制
发表时间：
2019-02-28
影响因子：
4.6
通讯作者：
Wada, Jun
中科院分区：
文献类型：
--
作者：
Hiramatsu, Sumie;Watanabe, Katsue S.;Wada, Jun
系统性红斑狼疮(SLE)患者外周血中CD4+细胞DNA整体甲基化水平降低可能在SLE发病机制中起关键作用。为了识别新的甲基化敏感基因，我们整合了MRL/LPR(MRL)和C57BL6/J(B6)小鼠的全基因组DNA甲基化和mRNA谱数据。我们鉴定了组织蛋白酶E(CTSE)，其中内含子1 583bp区域的13个甲基-CPGS发生了低甲基化，与B6小鼠相比，MRL小鼠CTSE基因表达上调。在B6小鼠中，mCGCG有93.3+/-2.05%甲基化，而在MRL小鼠中，有80.0+/-6.2%甲基化并突变为CGGG。已知Kaiso与mCGCG结合，我们假设它抑制B6小鼠CTSE的表达。芯片聚合酶链式反应显示，在MRL小鼠中，Kaiso与mCGCG结合减少。5-azaC和/或曲古菌素A处理的EL4细胞在芯片-聚合酶链式反应中显示Kaiso与mCGCG基序的结合受到抑制，而在qPCR中发现CTSE的过度表达。在EL4细胞中，siRNA沉默CTSE基因导致IL-10分泌减少。MCGCG基序低甲基化、Kaiso募集减少、CD4+细胞CTSE和IL-10表达增加可能参与了SLE的发病机制。
Global DNA hypomethylation in CD4+ cells in systemic lupus erythematosus (SLE) was suggested to play a key role in the pathogenesis. To identify new methylation-sensitive genes, we integrated genome-wide DNA methylation and mRNA profiling data in CD4+ cells of MRL/lpr (MRL) and C57BL6/J (B6) mice. We identified Cathepsin E (Ctse), in which 13 methyl-CpGs within 583 bp region of intron 1 were hypomethylated, and Ctse mRNA upregulated in MRL compared with B6 mice. One of methyl-CpGs, mCGCG was 93.3 +/- 2.05% methylated in B6 mice, while 80.0 +/- 6.2% methylated and mutated to CGGG in MRL mice. Kaiso is known to bind to mCGCG and we hypothesized that it represses expression of Ctse in B6 mice. The binding of Kaiso to mCGCG site in B6 mice was reduced in MRL mice revealed by ChIP-PCR. EL4 cells treated with 5-azaC and/or Trichostatin A showed the suppression of binding of Kaiso to mCGCG motif by ChIP-PCR and the overexpression of Ctse was demonstrated by qPCR. Ctse gene silencing by siRNA in EL4 cells resulted in reduction of IL-10 secretion. The hypomethylation of mCGCG motif, reduced recruitment of Kaiso, and increased expression of Ctse and Il-10 in CD4+ cells may be involved in the pathogenesis of SLE.