Kubaski Francyne, Brusius-Facchin Ana Carolina, Mason Robert W, Patel Pravin, Burin Maira G, Michelin-Tirelli Kristiane, Kessler Rejane Gus, Bender Fernanda, Leistner-Segal Sandra, Moreno Carolina A, Cavalcanti Denise P, Giugliani Roberto, Tomatsu Shunji
Nemours/Alfred I. duPont Hospital for Children, Wilmington, DE, USA.
Department of Biological Sciences, University of Delaware, Newark, DE, USA.
Prenat Diagn. 2017 May;37(5):435-439. doi: 10.1002/pd.5028. Epub 2017 Mar 12.
The aim of this study was to quantify glycosaminoglycans (GAGs) in amniotic fluid (AF) from an MPS VII fetus compared with age-matched fetuses obtained from normal pregnancies.
Disaccharides were measured by liquid chromatography tandem mass spectrometry, compared to age-matched controls. Enzyme assay was performed in AF supernatant or cultured amniocytes. GUSB was analyzed by next generation sequencing using Ion Torrent Personal Genome Machine with a customized panel.
No activity of β-glucuronidase was detected in fetal cells. The pregnancy was spontaneously terminated in the third trimester. Genetic studies identified a homozygous mutation of p.N379D (c.1135A > G) in the GUSB gene. Liquid chromatography tandem mass spectrometry showed that chondroitin sulfate, dermatan sulfate, heparan sulfate, and keratan sulfate levels were markedly increased in the MPS VII AF, compared to those in age-matched control AF (dermatan sulfate, heparan sulfate, and chondroitin-6-sulfate more than 10 × than age-matched controls; chondroitin-4-sulfate and keratan sulfate more than 3 times higher).
This is the first report of specific GAG analysis in AF from an MPS VII fetus, indicating that GAG elevation in AF occurs by 21 weeks of gestation and could be an additional tool for prenatal diagnosis of MPS VII and potentially other MPS types. © 2017 John Wiley & Sons, Ltd.
本研究旨在对黏多糖贮积症VII型（MPS VII）胎儿羊水（AF）中的糖胺聚糖（GAGs）进行定量分析，并与正常妊娠中年龄匹配的胎儿羊水进行比较。
采用液相色谱串联质谱法测定二糖，并与年龄匹配的对照组进行比较。在羊水上清液或培养的羊水中进行酶活性测定。使用Ion Torrent个人基因组机器和定制面板，通过下一代测序分析β-葡萄糖醛酸酶（GUSB）。
在胎儿细胞中未检测到β-葡萄糖醛酸酶活性。妊娠在孕晚期自然终止。基因研究在GUSB基因中鉴定出p.N379D（c.1135A＞G）纯合突变。液相色谱串联质谱分析显示，与年龄匹配的对照羊水相比，MPS VII胎儿羊水中硫酸软骨素、硫酸皮肤素、硫酸乙酰肝素和硫酸角质素水平显著升高（硫酸皮肤素、硫酸乙酰肝素和硫酸软骨素-6-硫酸酯比年龄匹配的对照组高10倍以上；硫酸软骨素-4-硫酸酯和硫酸角质素比对照组高3倍以上）。
这是首次对MPS VII胎儿羊水进行特异性GAG分析的报告，表明妊娠21周时羊水中GAG水平升高，这可能是MPS VII及其他潜在黏多糖贮积症类型产前诊断的一项辅助手段。©2017约翰威立父子有限公司
