TY - JOUR
T1 - Regulation of Cathepsin E gene expression by the transcription factor Kaiso in MRL/lpr mice derived CD4+ T cells
AU - Hiramatsu, Sumie
AU - Watanabe, Katsue S.
AU - Zeggar, Sonia
AU - Asano, Yosuke
AU - Miyawaki, Yoshia
AU - Yamamura, Yuriko
AU - Katsuyama, Eri
AU - Katsuyama, Takayuki
AU - Watanabe, Haruki
AU - Takano-Narazaki, Mariko
AU - Matsumoto, Yoshinori
AU - Kawabata, Tomoko
AU - Sada, Ken Ei
AU - Wada, Jun
N1 - Funding Information:
We thank Ms. Yoshiko Hada at Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences and Ms. Sumire Ishihara at Department of Animal Resources, Advanced Science Research Center, Okayama University for excellent technical support. Funding: This work was supported by JSPS Grant-in-Aid for Scientific Research, Grant numbers (16K09895, 16K09896, 16K19601, 16K19602, 16K19600, 17K09976, 18K16151).
Funding Information:
Competing Interests: K.S. receives speaker honoraria from Chugai Pharma. J.W. receives speaker honoraria from Daiichi Sankyo, M.S.D., Tanabe Mitsubishi, Taisho Toyama and receives grant support from Baxter, Dainippon Sumitomo, Ono, and Teijin Pharma.
PY - 2019/2/28
Y1 - 2019/2/28
N2 - 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.
AB - 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.
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U2 - 10.1038/s41598-019-38809-y
DO - 10.1038/s41598-019-38809-y
M3 - Article
C2 - 30816218
AN - SCOPUS:85062407065
VL - 9
SP - 3054
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 3054
ER -