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Article

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

Denali Therapeutics, Inc., 161 Oyster Point Blvd, South San Francisco, CA 94080, USA
*
Author to whom correspondence should be addressed.
Those authors contributed equally.
Present Address: Eisai G2D2, 35 Cambridge park Drive, Cambridge, MA 02141, USA.
Int. J. Mol. Sci. 2020, 21(15), 5449; https://doi.org/10.3390/ijms21155449
Received: 27 June 2020 / Revised: 27 July 2020 / Accepted: 28 July 2020 / Published: 30 July 2020
(This article belongs to the Special Issue Mucopolysaccharidoses: Diagnosis, Treatment, and Management)
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. View Full-Text
Keywords: mucopolysaccharidosis; iduronate 2-sulfatase; liquid chromatography-tandem mass spectrometry; glycosaminoglycans; heparan sulfate; dermatan sulfate; cerebrospinal fluid mucopolysaccharidosis; iduronate 2-sulfatase; liquid chromatography-tandem mass spectrometry; glycosaminoglycans; heparan sulfate; dermatan sulfate; cerebrospinal fluid
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MDPI and ACS Style

Wang, J.; Bhalla, A.; Ullman, J.C.; Fang, M.; Ravi, R.; Arguello, A.; Thomsen, E.; Tsogtbaatar, B.; Guo, J.L.; Skuja, L.L.; Dugas, J.C.; Davis, S.S.; Poda, S.B.; Gunasekaran, K.; Costanzo, S.; Sweeney, Z.K.; Henry, A.G.; Harris, J.M.; Henne, K.R.; Astarita, G. High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II. Int. J. Mol. Sci. 2020, 21, 5449. https://doi.org/10.3390/ijms21155449

AMA Style

Wang J, Bhalla A, Ullman JC, Fang M, Ravi R, Arguello A, Thomsen E, Tsogtbaatar B, Guo JL, Skuja LL, Dugas JC, Davis SS, Poda SB, Gunasekaran K, Costanzo S, Sweeney ZK, Henry AG, Harris JM, Henne KR, Astarita G. High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II. International Journal of Molecular Sciences. 2020; 21(15):5449. https://doi.org/10.3390/ijms21155449

Chicago/Turabian Style

Wang, Junhua, Akhil Bhalla, Julie C. Ullman, Meng Fang, Ritesh Ravi, Annie Arguello, Elliot Thomsen, Buyankhishig Tsogtbaatar, Jing L. Guo, Lukas L. Skuja, Jason C. Dugas, Sonnet S. Davis, Suresh B. Poda, Kannan Gunasekaran, Simona Costanzo, Zachary K. Sweeney, Anastasia G. Henry, Jeffrey M. Harris, Kirk R. Henne, and Giuseppe Astarita. 2020. "High-Throughput Liquid Chromatography–Tandem Mass Spectrometry Quantification of Glycosaminoglycans as Biomarkers of Mucopolysaccharidosis II" International Journal of Molecular Sciences 21, no. 15: 5449. https://doi.org/10.3390/ijms21155449

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