Gonçalves Margarida, Tillack Linda, de Carvalho Mamede, Pinto Susana, Conradt Harald S, Costa Júlia
Laboratory of Glycobiology, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal.
GlycoThera GmbH, Feodor-Lynen Strasse 35, 30625 Hannover, Germany.
Clin Chim Acta. 2015 Jan 1;438:342-9. doi: 10.1016/j.cca.2014.09.011. Epub 2014 Sep 28.
Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified.
We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry.
pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348).
Our results underscore the value of pNFH as a biomarker in ALS. In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker.
肌萎缩侧索硬化症(ALS)是一种运动神经元的致命性神经退行性疾病,目前尚未发现经过临床验证的生物标志物。
我们通过酶联免疫吸附测定法(ELISA)对29例ALS患者和19例年龄匹配的其他疾病对照患者脑脊液(CSF)中的生物标志物磷酸化神经丝重链(pNFH)进行了定量分析。此外,我们采用基于液相色谱和质谱的糖组学方法,比较了ALS患者和对照患者脑脊液中的蛋白质N-糖基化情况。
与对照组相比,ALS患者的pNFH水平显著更高(P<0.0001),尤其是在疾病快速进展者中。脑脊液中发现的N-聚糖主要是复杂的二天线型,带有α2,3-和α2,6-连接的唾液酸,以及含平分N-乙酰葡糖胺的结构和外周岩藻糖基化结构。与对照组相比,ALS组中由单唾液酸化二天线型聚糖A2G2S(6)1和FA2G2S(3)1组成的一个峰显著增加(P=0.0348)。
我们的结果强调了pNFH作为ALS生物标志物的价值。此外,我们发现了ALS患者N-糖基化模式的变化,表明这种变化应在未来研究中作为潜在生物标志物进行探索。