title: Identification of glioma neovascularization-related proteins by using MALDI-FTMS and nano-LC fractionation to microdissected tumor vessels.
abstract: The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.
authors.affiliation: Department of Pathology, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GD Rotterdam, The Netherlands.
authors.forename: Dana A N
authors.initials: DA
authors.lastname: Mustafa
authors.forename: Peter C
authors.initials: PC
authors.lastname: Burgers
authors.forename: Lennard J
authors.initials: LJ
authors.lastname: Dekker
authors.forename: Halima
authors.initials: H
authors.lastname: Charif
authors.forename: Mark K
authors.initials: MK
authors.lastname: Titulaer
authors.forename: Peter A E Sillevis
authors.initials: PA
authors.lastname: Smitt
authors.forename: Theo M
authors.initials: TM
authors.lastname: Luider
authors.forename: Johan M
authors.initials: JM
authors.lastname: Kros
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001808:Blood Vessels / Q000378:metabolism; D001932:Brain Neoplasms / Q000098:blood supply* / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D002853:Chromatography, Liquid; D005260:Female; D005340:Fibrinogen / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005910:Glioma / Q000098:blood supply* / Q000378:metabolism*; D006023:Glycoproteins; D006801:Humans; D007223:Infant; D008297:Male; D042282:Microdissection; D008875:Middle Aged; D036103:Nanotechnology; D009389:Neovascularization, Pathologic / Q000378:metabolism; D010455:Peptides / Q000032:analysis; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D014408:Biomarkers, Tumor; D002352:Carrier Proteins; D005353:Fibronectins; D006023:Glycoproteins; D010455:Peptides; C046289:colligin; D005340:Fibrinogen