title: Proteomics of vitreous in neovascular age-related macular degeneration.
abstract: Neovascular age-related macular degeneration (nAMD) has been described as a predominantly inflammatory and proangiogenic retino-choroidal disease. Vitreous humor (VH) is the adjacent and accessible compartment which, due to the vicinity to the retina, might best represent changes of protein-based mediators of nAMD. The aim of this clinical-experimental study was to analyze the nAMD associated VH proteome of previously untreated patients whilst taking different groups of nAMD into account, based on their clinical presentation (clinical diagnosis groups). Electrophoresis coupled online to mass spectrometry (CE-MS) as well as liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) were used to analyze VH of 108 nAMD patients and 24 controls with idiopathic floaters. A total of 101 different proteins with at least two unique peptides could be identified. Using a stringent statistical analysis with implementation of the closed test principle, we were able to identify four proteins that may be involved in the pathophysiology of nAMD: Clusterin, opticin, pigment epithelium-derived factor and prostaglandin-H2 d-isomerase. Using independent samples, ROC-Area under the curve was determined proving the validity of the results: Clusterin 0.747, opticin 0.656, pigment epithelium-derived factor 0.514, prostaglandin-H2 d-isomerase 0.712. In addition, validation through ELISA measurements was performed. The identified proteins may serve as potential biomarkers or even targets of therapy for nAMD.
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Nobl
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany; Department of Ophthalmology, University of Freiburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Reich
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany.
authors.forename: Ivanka
authors.initials: I
authors.lastname: Dacheva
authors.affiliation: Mosaiques Diagnostics GmbH, Hannover, Germany.
authors.forename: Justyna
authors.initials: J
authors.lastname: Siwy
authors.affiliation: BHF Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow, UK.
authors.forename: William
authors.initials: W
authors.lastname: Mullen
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France; Université Toulouse III Paul-Sabatier, Toulouse, France.
authors.forename: Joost P
authors.initials: JP
authors.lastname: Schanstra
authors.affiliation: Department of Ophthalmology, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; David J Apple International Laboratory for Ocular Pathology and International Vision Correction Research Centre (IVCRC), Heidelberg, Germany.
authors.forename: Chul Young
authors.initials: CY
authors.lastname: Choi
authors.affiliation: Department of Pathology, University of Heidelberg, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Kopitz
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany.
authors.forename: Florian T A
authors.initials: FTA
authors.lastname: Kretz
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany; David J Apple International Laboratory for Ocular Pathology and International Vision Correction Research Centre (IVCRC), Heidelberg, Germany.
authors.forename: Gerd U
authors.initials: GU
authors.lastname: Auffarth
authors.affiliation: Department of Ophthalmology, Goethe University, Frankfurt am Main, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Koch
authors.affiliation: Department of Ophthalmology, University of Heidelberg, Germany; David J Apple International Laboratory for Ocular Pathology and International Vision Correction Research Centre (IVCRC), Heidelberg, Germany; Department of Ophthalmology, University of Southern California, Los Angeles, CA, USA. Electronic address: Michael.koss@med.uni-heidelberg.de.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Koss
mesh_terms: D000368:Aged; D015415:Biomarkers / Q000378:metabolism; D020256:Choroidal Neovascularization / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D051152:Clusterin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D054834:Lipocalins / Q000378:metabolism*; D008268:Macular Degeneration / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D040901:Proteomics / Q000379:methods*; D012189:Retrospective Studies; D014822:Vitreous Body / Q000378:metabolism* / Q000473:pathology
chemical_list: D015415:Biomarkers; D051152:Clusterin; D054834:Lipocalins; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase
keywords: Biomarker; Clusterin; Neovascular age-related macular degeneration; Opticin; Pigment epithelium-derived factor; Prostaglandin-H2 d-isomerase; Proteomics; Vitreous humor