DNA Methylation Status of SHOX-Flanking CpG Islands in Healthy Individuals and Short Stature Patients with Pseudoautosomal Copy Number Variations
DNA Methylation Status of SHOX-Flanking CpG Islands in Healthy Individuals and Short Stature Patients with Pseudoautosomal Copy Number Variations
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DOI：
10.1159/000500468
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发表时间：
2019-06
影响因子：
1.7
通讯作者：
Kenichiro Ogushi;Atsushi Hattori;E. Suzuki;H. Shima;M. Izawa;Hideaki Yagasaki;R. Horikawa;K. Uetake;A. Umezawa;T. Ishii;K. Muroya;N. Namba;Toshiaki Tanaka;Yasuhiro Hirano;Hitoshi Yamamoto;S. Soneda;K. Matsubara;M. Kagami;M. Miyado;M. Fukami
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作者：
Kenichiro Ogushi;Atsushi Hattori;E. Suzuki;H. Shima;M. Izawa;Hideaki Yagasaki;R. Horikawa;K. Uetake;A. Umezawa;T. Ishii;K. Muroya;N. Namba;Toshiaki Tanaka;Yasuhiro Hirano;Hitoshi Yamamoto;S. Soneda;K. Matsubara;M. Kagami;M. Miyado;M. Fukami
Shox位于性染色体的短臂假常染色体区域(PAR1)，可以逃避X的失活。Shox单倍体功能不全是特发性矮小(ISS)和Leri-Weill软骨骨病(LWD)的基础。相当大比例的Shox单倍体缺陷病例是由涉及Shox基因增强子区域的伪常染色体拷贝数变异(CNV)引起的。我们先前使用外周血样本的研究表明，在一名健康女性中，Shox外显子1附近的一些CpG二核苷酸发生了低甲基化，而在一名X染色体大重排患者中发生了甲基化。然而，目前尚不清楚亚显微伪常染色体CNV是否会导致Shox侧翼CpG岛的异常DNA甲基化。在这项研究中，我们检测了50名健康人和10名ISS/LWD假常染色体CNV患者Shox侧翼CpG岛的DNA甲基化状态。在电子计算机分析中，在Shox的翻译起始点的20kb区域内检测到3个CpG岛。基因组DNA焦解测序和亚硫酸氢盐测序显示，这些CpG岛在健康人的外周血细胞和培养的软骨细胞中几乎没有甲基化，在ISS/LWD假常染色体CNV患者的外周血细胞中也几乎没有甲基化。这些结果，结合我们以前的发现，表明Shox侧翼CpG岛的DNA甲基化状态可以受到X染色体总体异常的影响，但不会受到PAR1中亚微观CNV的影响。这些CNV可能通过DNA甲基化独立的机制干扰Shox的表达，这需要在未来的研究中确定。
SHOX resides in the short arm pseudoautosomal region (PAR1) of the sex chromosomes and escapes X inactivation. SHOX haploinsufficiency underlies idiopathic short stature (ISS) and Leri-Weill dyschondrosteosis (LWD). A substantial percentage of cases with SHOX haploinsufficiency arise from pseudoautosomal copy number variations (CNVs) involving putative enhancer regions of SHOX. Our previous study using peripheral blood samples showed that some CpG dinucleotides adjacent to SHOX exon 1 were hypomethylated in a healthy woman and methylated in a woman with gross X chromosomal rearrangements. However, it remains unknown whether submicroscopic pseudoautosomal CNVs cause aberrant DNA methylation of SHOX-flanking CpG islands. In this study, we examined the DNA methylation status of SHOX-flanking CpG islands in 50 healthy individuals and 10 ISS/LWD patients with pseudoautosomal CNVs. In silico analysis detected 3 CpG islands within the 20-kb region from the translation start site of SHOX. Pyrosequencing and bisulfite sequencing of genomic DNA samples revealed that these CpG islands were barely methylated in peripheral blood cells and cultured chondrocytes of healthy individuals, as well as in peripheral blood cells of ISS/LWD patients with pseudoautosomal CNVs. These results, in conjunction with our previous findings, indicate that the DNA methylation status of SHOX-flanking CpG islands can be affected by gross X-chromosomal abnormalities, but not by submicroscopic CNVs in PAR1. Such CNVs likely disturb SHOX expression through DNA methylation-independent mechanisms, which need to be determined in future studies.