GlycoFibroTest Is a Highly Performant Liver Fibrosis Biomarker Derived from DNA Sequencer-based Serum Protein Glycomics
GlycoFibroTest Is a Highly Performant Liver Fibrosis Biomarker Derived from DNA Sequencer-based Serum Protein Glycomics
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DOI：
10.1074/mcp.m800470-mcp200
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发表时间：
2009-05-01
影响因子：
7
通讯作者：
Callewaert, Nico
中科院分区：
文献类型：
--
作者：
Vanderschaeghe, Dieter;Laroy, Wouter;Callewaert, Nico
目前肝纤维化的评估是通过肝活检来进行的，这是一种昂贵且相当繁琐的程序，不适合频繁的患者监测，这推动了对生物标记物的研究。为了研究血清N-糖是否包含适合这一目标的信息，我们开发了一种基于96孔板的血清N-糖样品制备方法，该方法只涉及液体转移步骤和在聚合酶链式反应热循环仪中孵育产生8-氨基比林-1，3，6-三磺酸标记的N-糖链。然后，这些N-葡聚糖就可以在基于毛细管电泳法的DNA测序仪上进行分析，这是目前世界各地临床遗传学实验室的标准。随后，我们对连续376名慢性丙型肝炎病毒患者进行了一项多中心、盲法研究，这些患者的肝脏活检和广泛的血清生化数据是可用的。在患者中，METAVIR纤维化分期分布如下：10.6%F0，44.4%F1，20.5%F2，18.4%F3，6.1%F4。我们发现，两个N-糖链的比率，这里称为GlycoFibroTest，与目前临床上使用的FibroTest(F1-F4中的Rho=0.4-0.5)一样与组织学纤维化阶段相关。最后，使用亲和层析，我们耗尽了血清中免疫球蛋白G，这导致了从N-糖基中完全去除了低半乳糖化的双天线多糖，这是部分确定GlycoFibroTest的。《分子与细胞蛋白质组学》8：986-994,2009。
Liver fibrosis is currently assessed by liver biopsy, a costly and rather cumbersome procedure that is unsuitable for frequent patient monitoring, which drives research into biomarkers for this purpose. To investigate whether the serum N-glycome contains information suitable for this goal, we developed a 96-well plate-based serum N-glycomics sample preparation protocol that only involves fluid transfer steps and incubations in a PCR thermocycler yielding 8-aminopyrene-1,3,6-trisulfonic acid-labeled N-glycans. These N-glycans are then ready for analysis on the capillary electrophoresis-based DNA sequencers that are the current standard in clinical genetics laboratories worldwide. Subsequently we performed a multicenter, blinded study of 376 consecutive chronic hepatitis C virus patients for which liver biopsies and extensive serum biochemistry data were available. Among patients, the METAVIR fibrosis stage distribution was as follows: 10.6% F0, 44.4% F1, 20.5% F2, 18.4% F3, and 6.1% F4. We found that the ratio of two N-glycans, here called GlycoFibroTest, correlates with the histological fibrosis stage equally well as FibroTest (rho = 0.4-0.5 in F1-F4), which is used in the clinic today. Finally using affinity chromatography we depleted sera of immunoglobulin G, and this resulted in a complete removal of the undergalactosylated biantennary glycans from the N-glycome, which are partially determining GlycoFibroTest. Molecular & Cellular Proteomics 8: 986-994, 2009.