Genome-wide DNA methylation profiling of hip articular cartilage identifies differentially methylated loci associated with osteonecrosis of the femoral head
Genome-wide DNA methylation profiling of hip articular cartilage identifies differentially methylated loci associated with osteonecrosis of the femoral head
复制标题
髋关节软骨的全基因组 DNA 甲基化谱鉴定了与股骨头坏死相关的差异甲基化位点
DOI：
10.1016/j.bone.2019.06.021
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发表时间：
2019-10-01
期刊：
影响因子：
4.1
通讯作者：
Liu, Ruiyu
中科院分区：
文献类型：
--
作者：
Wu, Junlong;Du, Yanan;Liu, Ruiyu
目的：最近的研究表明，髋关节软骨破坏在股骨头坏死(ONFH)的发生发展中起着重要作用。方法：收集15例股骨颈骨折患者(男11例，女4例)和15例对照组(男11例，女4例)股骨颈骨折患者髋关节软骨标本。ONFH患者和对照组的平均年龄分别为50.27±5.27岁和61.67±3.38岁。用Illumina Human Megylation850芯片检测5个ONFH和5个对照软骨的全基因组DNA甲基化图谱。DNA甲基化图谱的差异甲基化分析通过对LIMMA包进行经验性贝叶斯调节的t检验进行。对10个ONFH软骨和10个正常软骨进行质谱仪(MS)分析，以验证全基因组DNA甲基化图谱的结果。用免疫组织化学方法检测4例ONFH软骨和4例对照软骨中差异甲基化基因编码蛋白的表达水平。结果：共发现2872个差异甲基化的CpG位点，分别注释为480个高甲基化基因和1335个低甲基化基因。MS验证的结果与全基因组DNA甲基化图谱的结果一致。免疫组化进一步证实CARS(浅层59.67%[48.46，56.14]，深层31%[25.85，30.61])，PDE4D(浅层50.33%[33.64，40.68]，深层28.67%[10.81，36.47])，ADAMTS12(浅层53.67%[36.01，40.93]，深层34.67%[22.56，37.181])，LRP5(浅层59.63%[27.32，39.61]，深层22.95%[5.28，19.29])，RUNX2(浅层52.58%[11.64，31.33]，深层35.01%[10.03，27.44])。结论：DNA甲基化改变在ONFH的发生发展中起重要作用，为ONFH的发病机制和治疗研究提供了新的线索。
Objective: Recent studies demonstrated a critical role of hip articular cartilage destruction in the development of osteonecrosis of the femoral head (ONFH). The aim of this study was to characterize the genome-wide DNA methylation profile of hip cartilage obtained from patients with ONFH and healthy subjects.Methods: Hip articular cartilage specimens were collected from 15 ONFH patients (including 11 males and 4 females) and 15 control subjects (including 11 males and 4 females) with femoral neck fracture. The average ages of the ONFH patients and control subjects were 50.27 +/- 5.27 years and 61.67 +/- 3.38 years, respectively. Genome-wide DNA methylation profiles of 5 ONFH and 5 control cartilages were determined by Illumina HumanMethylation850 array. Differential methylation analysis of DNA methylation profiles were performed by the empirical Bayes moderated t-test of the limma package. Mass spectrograph (MS) analysis of 10 ONFH cartilages and 10 normal cartilages were performed to validate the results of genome-wide DNA methylation profiling. Immunohistochemistry (IHC) of 4 ONFH cartilages and 4 control cartilages were conducted to evaluate the expression levels of proteins encoded by identified differentially methylated genes. t-test was used to assess the significance of protein expression differences between ONFH patients and controls in IHC.Results: We identified a total of 2872 differentially methylated CpG sites, annotated to 480 hypermethylated genes and 1335 hypomethylated genes for ONFH. The results of MS validation were consistent with that of genome-wide DNA methylation profiling. IHC further confirmed the increased protein expression of CARS (mean and 95%CI of superficial zone 59.67% [48.46, 56.14], and deep zone 31% [25.85, 30.61]), PDE4D (superficial zone 50.33% [33.64, 40.68] and deep zone 28.67% [10.81, 36.47]), ADAMTS12 (superficial zone 53.67% [36.01, 40.93] and deep zone 34.67% [22.56, 37.181), LRP5 (superficial zone 59.63% [27.32, 39.61] and deep zone 22.95% [5.28, 19.29]), RUNX2 (superficial zone 52.58% [11.64, 31.33] and deep zone 35.01% [10.03, 27.44]) in ONFH articular cartilage.Conclusion: Our results suggest the implication of DNA methylation alterations in the development of ONFH, and provide novel clues for pathogenetic and therapeutic studies of ONFH.