Kuchar Ladislav, Berna Linda, Poupetova Helena, Ledvinova Jana, Ruzicka Petr, Dostalova Gabriela, Reichmannova Stella, Asfaw Befekadu, Linhart Ales, Sikora Jakub
Research Unit for Rare Diseases, Department of Paediatrics and Inherited Metabolic Disorders, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.
Research Unit for Rare Diseases, Department of Paediatrics and Inherited Metabolic Disorders, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.
Clin Chim Acta. 2024 Jul 15;561:119824. doi: 10.1016/j.cca.2024.119824. Epub 2024 Jun 19.
Fabry disease (FD) is an X-linked lysosomal storage disease resulting from pathogenic variants in the GLA gene coding α-galactosidase A (AGAL) and cleaving terminal alpha-linked galactose. Globotriaosylceramide (Gb3) is the predominantly accumulated sphingolipid. Gb3, deacylated-Gb3 (lysoGb3), and methylated-Gb3 (metGb3) have been suggested as FD biomarkers.
We developed a novel LC-MS/MS method for assessing lysoGb3 levels in plasma and Gb3 and metGb3 in urine and tested 62 FD patients, 34 patients with GLA variants of unknown significance (VUS) and 59 healthy controls. AGAL activity in white blood cells (WBCs) and plasma was evaluated in parallel.
In males, lysoGb3 concentrations in plasma separated classic and late-onset FD patients from each other and from individuals carrying GLA VUS and healthy controls. Calculating AGAL activity/plasmatic lysoGb3 ratio allowed to correctly categorize all females with classic and majority of patients with late-onset FD phenotypes. Correlation of AGAL activity in WBCS with lipid biomarkers identified threshold activity values under which the biomarkers' concentrations increase.
We developed a novel simplified LC-MS/MS method for quantitation of plasma lysoGb3. AGAL activity/plasma lysoGb3 ratio was identified as the best predictor for FD. AGAL activity correlated with plasma lysoGb3 and corresponded to individual FD phenotypes.
法布里病（FD）是一种X连锁溶酶体贮积病，由编码α-半乳糖苷酶A（AGAL）并切割末端α-连接半乳糖的GLA基因中的致病变异引起。Globotriaosylceramide（Gb3）是主要蓄积的鞘脂。Gb3、脱酰基-Gb3（lysoGb3）和甲基化-Gb3（metGb3）已被提议作为法布里病的生物标志物。
我们开发了一种新型液相色谱-串联质谱法，用于评估血浆中的lysoGb3水平以及尿液中的Gb3和metGb3水平，并对62例法布里病患者、34例意义未明的GLA变异（VUS）患者和59名健康对照进行了检测。同时评估了白细胞（WBC）和血浆中的AGAL活性。
在男性中，血浆中的lysoGb3浓度将经典型和晚发型法布里病患者彼此区分开来，并与携带GLA VUS的个体和健康对照区分开来。计算AGAL活性/血浆lysoGb3比值能够正确分类所有经典型女性患者和大多数晚发型法布里病表型患者。白细胞中的AGAL活性与脂质生物标志物的相关性确定了生物标志物浓度升高时的阈值活性值。
我们开发了一种新型简化的液相色谱-串联质谱法用于定量血浆lysoGb3。AGAL活性/血浆lysoGb3比值被确定为法布里病的最佳预测指标。AGAL活性与血浆lysoGb3相关，并与个体法布里病表型相对应。
