title: Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis.
abstract: BACKGROUND
  Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified.
  
  
  METHODS
  We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry.
  
  
  RESULTS
  pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348).
  
  
  CONCLUSIONS
  Our results underscore the value of pNFH as a biomarker in ALS. In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker.
authors.affiliation: Laboratory of Glycobiology, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal.
authors.forename: Margarida
authors.initials: M
authors.lastname: Gonçalves
authors.affiliation: GlycoThera GmbH, Feodor-Lynen Strasse 35, 30625 Hannover, Germany.
authors.forename: Linda
authors.initials: L
authors.lastname: Tillack
authors.affiliation: Department Neurosciences, Hospital de Santa Maria, Lisbon, Portugal; Translational Clinical Physiology Unit, Instituto de Medicina Molecular, Institute of Physiology, Faculty of Medicine - University of Lisbon, Portugal.
authors.forename: Mamede
authors.initials: M
authors.lastname: de Carvalho
authors.affiliation: Department Neurosciences, Hospital de Santa Maria, Lisbon, Portugal; Translational Clinical Physiology Unit, Instituto de Medicina Molecular, Institute of Physiology, Faculty of Medicine - University of Lisbon, Portugal.
authors.forename: Susana
authors.initials: S
authors.lastname: Pinto
authors.affiliation: GlycoThera GmbH, Feodor-Lynen Strasse 35, 30625 Hannover, Germany.
authors.forename: Harald S
authors.initials: HS
authors.lastname: Conradt
authors.affiliation: Laboratory of Glycobiology, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal. Electronic address: jcosta@itqb.unl.pt.
authors.forename: Júlia
authors.initials: J
authors.lastname: Costa
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry / Q000302:isolation & purification; D000328:Adult; D000368:Aged; D000690:Amyotrophic Lateral Sclerosis / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D015415:Biomarkers / Q000134:cerebrospinal fluid; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D018450:Disease Progression; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005643:Fucose / Q000737:chemistry / Q000302:isolation & purification; D054794:Glycomics / Q000295:instrumentation / Q000379:methods; D006031:Glycosylation; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D016900:Neurofilament Proteins / Q000134:cerebrospinal fluid*; D010750:Phosphoproteins / Q000134:cerebrospinal fluid*; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification; D012794:Sialic Acids / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D015415:Biomarkers; D016900:Neurofilament Proteins; D010750:Phosphoproteins; D011134:Polysaccharides; D012794:Sialic Acids; C051774:neurofilament protein H; D005643:Fucose; D000117:Acetylglucosamine
keywords: Amyotrophic lateral sclerosis; Biomarker; Cerebrospinal fluid; Glycomics; Glycoproteins; Phosphoneurofilament heavy chain