High-Throughput Liquid Chromatography-Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II

被引：11

作者：

Wang, Junhua ^{[1
]}

Bhalla, Akhil ^{[1
]}

Ullman, Julie C. ^{[1
]}

Fang, Meng ^{[1
]}

Ravi, Ritesh ^{[1
]}

Arguello, Annie ^{[1
]}

Thomsen, Elliot ^{[1
]}

Tsogtbaatar, Buyankhishig ^{[1
]}

Guo, Jing L. ^{[1
]}

Skuja, Lukas L. ^{[1
]}

Dugas, Jason C. ^{[1
]}

Davis, Sonnet S. ^{[1
]}

Poda, Suresh B. ^{[1
,2
]}

Gunasekaran, Kannan ^{[1
]}

Costanzo, Simona ^{[1
]}

Sweeney, Zachary K. ^{[1
]}

Henry, Anastasia G. ^{[1
]}

Harris, Jeffrey M. ^{[1
]}

Henne, Kirk R. ^{[1
]}

Astarita, Giuseppe ^{[1
]}

机构：

[1] Denali Therapeut Inc, 161 Oyster Point Blvd, San Francisco, CA 94080 USA

[2] Eisai G2D2, 35 Cambridge Pk Dr, Cambridge, MA 02141 USA

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2020年 / 21卷 / 15期

关键词：

mucopolysaccharidosis; iduronate; 2-sulfatase; liquid chromatography-tandem mass spectrometry; glycosaminoglycans; heparan sulfate; dermatan sulfate; cerebrospinal fluid; HEPARAN-SULFATE; CEREBROSPINAL-FLUID; CHONDROITIN SULFATE; DERMATAN SULFATE; DISACCHARIDES; URINE; ANTIBODY; CSF;

D O I：

10.3390/ijms21155449

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We recently developed a blood-brain barrier (BBB)-penetrating enzyme transport vehicle (ETV) fused to the lysosomal enzyme iduronate 2-sulfatase (ETV:IDS) and demonstrated its ability to reduce glycosaminoglycan (GAG) accumulation in the brains of a mouse model of mucopolysaccharidosis (MPS) II. To accurately quantify GAGs, we developed a plate-based high-throughput enzymatic digestion assay coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to simultaneously measure heparan sulfate and dermatan sulfate derived disaccharides in tissue, cerebrospinal fluid (CSF) and individual cell populations isolated from mouse brain. The method offers ultra-high sensitivity enabling quantitation of specific GAG species in as low as 100,000 isolated neurons and a low volume of CSF. With an LOD at 3 ng/mL and LLOQs at 5-10 ng/mL, this method is at least five times more sensitive than previously reported approaches. Our analysis demonstrated that the accumulation of CSF and brain GAGs are in good correlation, supporting the potential use of CSF GAGs as a surrogate biomarker for brain GAGs. The bioanalytical method was qualified through the generation of standard curves in matrix for preclinical studies of CSF, demonstrating the feasibility of this assay for evaluating therapeutic effects of ETV:IDS in future studies and applications in a wide variety of MPS disorders.

引用

页码：1 / 17

页数：17

共 44 条

[41] Glycosaminoglycans in human cerebrospinal fluid determined by LC-MS/MS MRM [J].

Yu, Yanlei ;

Zhang, Fuming ;

Colon, Wilfredo ;

Linhardt, Robert J. ;

Xia, Ke .

ANALYTICAL BIOCHEMISTRY, 2019, 567 :82-84

[42] A straightforward, quantitative ultra-performance liquid chromatography-tandem mass spectrometric method for heparan sulfate, dermatan sulfate and chondroitin sulfate in urine: An improved clinical screening test for the mucopolysaccharidoses [J].

Zhang, Haoyue ;

Wood, Tim ;

Young, Sarah P. ;

Millington, David S. .

MOLECULAR GENETICS AND METABOLISM, 2015, 114 (02) :123-128

[43] Analysis of Glycosaminoglycans in Cerebrospinal Fluid from Patients with Mucopolysaccharidoses by Isotope-Dilution Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry [J].

Zhang, Haoyue ;

Young, Sarah P. ;

Auray-Blais, Christiane ;

Orchard, Paul J. ;

Tolar, Jakub ;

Millington, David S. .

CLINICAL CHEMISTRY, 2011, 57 (07) :1005-1012

[44] Glycosaminoglycan (GAG) Biosynthesis and GAG-Binding Proteins [J].

Zhang, Lijuan .

GLYCOSAMINOGLYCANS IN DEVELOPMENT, HEALTH AND DISEASE, 2010, 93 :1-17

← 1 2 3 4 5 →