title: Proteomic Discovery and Development of a Multiplexed Targeted MRM-LC-MS/MS Assay for Urine Biomarkers of Extracellular Matrix Disruption in Mucopolysaccharidoses I, II, and VI.
abstract: The mucopolysaccharidoses (MPS) are lysosomal storage disorders that result from defects in the catabolism of glycosaminoglycans. Impaired muscle, bone, and connective tissue are typical clinical features of MPS due to disruption of the extracellular matrix. Markers of MPS disease pathology are needed to determine disease severity and monitor effects of existing and emerging new treatments on disease mechanisms. Urine samples from a small cohort of MPS-I, -II, and -VI patients (n = 12) were analyzed using label-free quantative proteomics. Fifty-three proteins including many associated with extracellular matrix organization were differently expressed. A targeted multiplexed peptide MRM LC-MS/MS assay was used on a larger validation cohort of patient samples (MPS-I n = 18, MPS-II n = 12, MPS-VI n = 6, control n = 20). MPS-I and -II groups were further subdivided according to disease severity. None of the markers assessed were altered significantly in the mild disease groups compared to controls. β-galactosidase, a lysosomal protein, was elevated 3.6-5.7-fold significantly (p < 0.05) in all disease groups apart from mild MPS-I and -II. Collagen type Iα, fatty-acid-binding-protein 5, nidogen-1, cartilage oligomeric matrix protein, and insulin-like growth factor binding protein 7 concentrations were elevated in severe MPS I and II groups. Cartilage oligomeric matrix protein, insulin-like growth factor binding protein 7, and β-galactosidase were able to distinguish the severe neurological form of MPS-II from the milder non-neurological form. Protein Heg1 was significantly raised only in MPS-VI. This work describes the discovery of new biomarkers of MPS that represent disease pathology and allows the stratification of MPS-II patients according to disease severity.
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Wendy E
authors.initials: WE
authors.lastname: Heywood
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Stephane
authors.initials: S
authors.lastname: Camuzeaux
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Ivan
authors.initials: I
authors.lastname: Doykov
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Nina
authors.initials: N
authors.lastname: Patel
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Rhian-Lauren
authors.initials: RL
authors.lastname: Preece
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Emma
authors.initials: E
authors.lastname: Footitt
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Maureen
authors.initials: M
authors.lastname: Cleary
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Clayton
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Grunewald
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Lara
authors.initials: L
authors.lastname: Abulhoul
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Anupam
authors.initials: A
authors.lastname: Chakrapani
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Sebire
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Hindmarsh
authors.affiliation: University of Groningen , University Medical Center Groningen, Departments of Genetics and Neurology, P.O. Box 30.001, 9700 RB Groningen, Netherlands.
authors.forename: Tom J
authors.initials: TJ
authors.lastname: de Koning
authors.affiliation: Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.|Enzyme and Metabolic Unit, Chemical Pathology, Great Ormond Street Hospital for Children , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Simon
authors.initials: S
authors.lastname: Heales
authors.affiliation: Enzyme and Metabolic Unit, Chemical Pathology, Great Ormond Street Hospital for Children , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Derek
authors.initials: D
authors.lastname: Burke
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.|Centre for Inborn Errors of Metabolism, Great Ormond Street Hospital , Great Ormond Street, London, WC1N 3JH, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Gissen
authors.affiliation: Centre for Translational Omics, UCL Institute of Child Health , 30 Guilford Street, London, WC1N 1EH United Kingdom.
authors.forename: Kevin
authors.initials: K
authors.lastname: Mills
mesh_terms: D015415:Biomarkers / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D005109:Extracellular Matrix / Q000378:metabolism*; D006801:Humans; D009083:Mucopolysaccharidoses / Q000378:metabolism* / Q000652:urine*; D036103:Nanotechnology; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers