Background

Urine and plasma biomarker testing for lysosomal storage disorders by liquid chromatography mass spectrometry (LC-MS) currently requires multiple analytical methods to detect the abnormal accumulation of oligosaccharides, mucopolysaccharides, and glycolipids. To improve clinical testing efficiency, we developed a single LC-MS method to simultaneously identify disorders of oligosaccharide, mucopolysaccharide, and glycolipid metabolism with minimal sample preparation.

Methods

We created a single chromatographic method for separating free glycans and glycolipids in their native form, using an amide column and high pH conditions. We used this glycomic profiling method both in untargeted analyses of patient and control urines using LC ion-mobility high-resolution MS (biomarker discovery), and targeted analyses of urine, serum, and dried blood spot samples by LC-MS/MS (clinical validation).

Results

Untargeted glycomic profiling revealed twenty biomarkers that could identify and subtype mucopolysaccharidoses. We incorporated these with known oligosaccharide and glycolipid biomarkers into a rapid test that identifies at least 27 lysosomal storage disorders, including oligosaccharidoses, mucopolysaccharidoses, sphingolipidoses, glycogen storage disorders, and congenital disorders of glycosylation and de-glycosylation. In a validation set containing 115 urine samples from patients with lysosomal storage disorders, all were unambiguously distinguished from normal controls, with correct disease subtyping for 88% (101/115) of cases. Glucosylsphingosine was reliably elevated in dried blood spots from Gaucher disease patients with baseline resolution from galactosylsphingosine.

Conclusion

Glycomic profiling by liquid chromatography mass spectrometry identifies a range of lysosomal storage disorders. This test can be used in clinical evaluations to rapidly focus a diagnosis, as well as to clarify or support additional gene sequencing and enzyme studies.

中文翻译：

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使用液相色谱质谱法通过糖组学分析检测溶酶体贮积症

背景

通过液相色谱质谱 (LC-MS) 检测溶酶体贮积症的尿液和血浆生物标志物目前需要多种分析方法来检测寡糖、粘多糖和糖脂的异常积累。为了提高临床测试效率，我们开发了一种单一的 LC-MS 方法，以最少的样品制备同时鉴定寡糖、粘多糖和糖脂代谢紊乱。

方法

我们创建了一种使用酰胺柱和高 pH 条件分离天然形式的游离聚糖和糖脂的单一色谱方法。我们在使用 LC 离子迁移率高分辨率 MS（生物标志物发现）对患者和对照尿液进行非靶向分析，以及通过 LC-MS/MS（临床验证）。

结果

非靶向糖组学分析揭示了二十种可以识别粘多糖症并对其进行亚型分析的生物标志物。我们将这些与已知的寡糖和糖脂生物标志物结合到一个快速测试中，该测试可识别至少 27 种溶酶体贮积症，包括寡糖贮积症、粘多糖贮积症、鞘脂贮积症、糖原贮积症以及糖基化和去糖基化的先天性疾病。在包含来自溶酶体贮积症患者的 115 份尿液样本的验证集中，所有样本都与正常对照明确区分开来，88% (101/115) 的病例具有正确的疾病亚型。葡萄糖基鞘氨醇在戈谢病患者的干血斑中可靠地升高，基线分辨率与半乳糖基鞘氨醇有关。

结论

通过液相色谱质谱法进行的糖组学分析确定了一系列溶酶体贮积症。该测试可用于临床评估以快速集中诊断，以及澄清或支持额外的基因测序和酶研究。