Genome-wide DNA methylation analysis of pseudohypoparathyroidism patients with GNAS imprinting defects.
Genome-wide DNA methylation analysis of pseudohypoparathyroidism patients with GNAS imprinting defects.
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DOI：
10.1186/s13148-016-0175-8
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发表时间：
2016
影响因子：
5.7
通讯作者：
Freson K
中科院分区：
文献类型：
--
作者：
Rochtus A;Martin-Trujillo A;Izzi B;Elli F;Garin I;Linglart A;Mantovani G;Perez de Nanclares G;Thiele S;Decallonne B;Van Geet C;Monk D;Freson K
假性甲状旁腺功能减退症（PHP）是由印迹GNAS簇中的（epi）遗传缺陷引起的。目前PHP患者的分类受到临床和分子诊断重叠的阻碍。欧洲PHP研究联盟设计了一项全基因组甲基化研究，以改善分子诊断。使用HumanMethylation 450 K BeadChip分析了24名甲状旁腺激素抵抗PHP患者和20名年龄和性别匹配的对照组的全基因组甲基化。患者先前被诊断患有GNAS特异性差异甲基化区域（DMR），包括6名已知STX 16缺失（PHPΔ stx 16）和18名无缺失（PHPneg）的患者。该阵列表明PHP患者在全基因组水平上不显示DNA甲基化差异。GNAS特异性DMR的无监督聚类将PHPΔ stx 16与PHPneg患者区分开来。有趣的是，与所有PHP患者在A/B DMR中共享甲基化缺陷而只有PHPΔ stx 16患者具有正常NESP，GNAS-AS 1和XL甲基化的观点相反，我们在GNAS-AS 1区域发现了一种新的DMR（命名为GNAS-AS 2），其在PHPΔ stx 16和PHPneg中均显着不同，如Sequenom EpiTYPER在更大的PHP队列中所验证的。对58例DMR的分析显示，8/18例PHPneg和16例PHPΔ stx 16患者存在多位点甲基化缺陷。对FANCC和SVOPL DMR进行了确认。这是第一个针对PHP患者的全基因组甲基化研究，证实GNAS是最重要的DMR，STX 16缺失的存在将PHP患者分为两组。此外，一种新的GNAS-AS 2 DMR影响所有PHP患者，PHP患者似乎对多位点甲基化缺陷敏感。本文的在线版本（doi：10.1186/s13148-016-0175-8）包含补充材料，可供授权用户使用。
Pseudohypoparathyroidism (PHP) is caused by (epi)genetic defects in the imprinted GNAS cluster. Current classification of PHP patients is hampered by clinical and molecular diagnostic overlaps. The European Consortium for the study of PHP designed a genome-wide methylation study to improve molecular diagnosis. The HumanMethylation 450K BeadChip was used to analyze genome-wide methylation in 24 PHP patients with parathyroid hormone resistance and 20 age- and gender-matched controls. Patients were previously diagnosed with GNAS-specific differentially methylated regions (DMRs) and include 6 patients with known STX16 deletion (PHPΔstx16) and 18 without deletion (PHPneg). The array demonstrated that PHP patients do not show DNA methylation differences at the whole-genome level. Unsupervised clustering of GNAS-specific DMRs divides PHPΔstx16 versus PHPneg patients. Interestingly, in contrast to the notion that all PHP patients share methylation defects in the A/B DMR while only PHPΔstx16 patients have normal NESP, GNAS-AS1 and XL methylation, we found a novel DMR (named GNAS-AS2) in the GNAS-AS1 region that is significantly different in both PHPΔstx16 and PHPneg, as validated by Sequenom EpiTYPER in a larger PHP cohort. The analysis of 58 DMRs revealed that 8/18 PHPneg and 1/6 PHPΔstx16 patients have multi-locus methylation defects. Validation was performed for FANCC and SVOPL DMRs. This is the first genome-wide methylation study for PHP patients that confirmed that GNAS is the most significant DMR, and the presence of STX16 deletion divides PHP patients in two groups. Moreover, a novel GNAS-AS2 DMR affects all PHP patients, and PHP patients seem sensitive to multi-locus methylation defects. The online version of this article (doi:10.1186/s13148-016-0175-8) contains supplementary material, which is available to authorized users.