A software application for comparing large numbers of high resolution MALDI-FTICR MS spectra demonstrated by searching candidate biomarkers for glioma blood vessel formation.
BACKGROUND
A Java application is presented, which compares large numbers (n > 100) of raw FTICR mass spectra from patients and controls. Two peptide profile matrices can be produced simultaneously, one with occurrences of peptide masses in samples and another with the intensity of common peak masses in all the measured samples, using the peak- and background intensities of the raw data. In latter way, more significantly differentially expressed peptides are found between groups than just using the presence or absence in samples of common peak masses. The software application is tested by searching angiogenesis related proteins in glioma by comparing laser capture micro dissected- and enzymatic by trypsin digested tissue sections.


RESULTS
By hierarchical clustering of the presence-absence matrix, it appears that proteins, such as hemoglobin alpha and delta subunit, fibrinogen beta and gamma chain precursor, tubulin specific chaperone A, epidermal fatty acid binding protein, neutrophil gelatinase-associated lipocalin precursor, peptidyl tRNA hydrolase 2 mitochondrial precursor, placenta specific growth hormone, and zinc finger CCHC domain containing protein 13 are significantly different expressed in glioma vessels. The up-regulated proteins in the glioma vessels with respect to the normal vessels determined by the Wilcoxon-Mann-Whitney test on the intensity matrix are vimentin, glial fibrillary acidic protein, serum albumin precursor, annexin A5, alpha cardiac and beta actin, type I cytoskeletal 10 keratin, calcium binding protein p22, and desmin. Peptide masses of calcium binding protein p22, Cdc42 effector protein 3, fibronectin precursor, and myosin-9 are exclusively present in glioma vessels. Some peptide fragments of non-muscular myosin-9 at the C-terminus are strongly up-regulated in the glioma vessels with respect to the normal vessels.


CONCLUSIONS
The less rigorous than in general used commercial propriety software de-isotope algorithm results in more mono-isotopic peptide masses and consequently more proteins. Centroiding of peptide masses takes place by taking the average over more spectra in the profile matrix. Cytoskeleton proteins and proteins involved in the calcium signaling pathway seem to be most up-regulated in glioma vessels. The finding that peptides at the C-terminus of myosin-9 are up-regulated could be ascribed to splicing or fragmentation by proteases.
Titulaer
Mark K
MK
Department of Neurology, Laboratory of Neuro-Oncology, Clinical and Cancer Proteomics, Erasmus Medical Center, Dr, Molewaterplein 50, PO Box 2040, 3000 CA Rotterdam, The Netherlands. m.titulaer@erasmusmc.nl
Mustafa
Dana A N
DA
Siccama
Ivar
I
Konijnenburg
Marco
M
Burgers
Peter C
PC
Andeweg
Arno C
AC
Smitt
Peter A E Sillevis
PA
Kros
Johan M
JM
Luider
Theo M
TM
D000465:Algorithms; D042501:Angiogenic Proteins / Q000032:analysis*; D014408:Biomarkers, Tumor / Q000032:analysis; D017357:Cyclotrons; D005910:Glioma / Q000098:blood supply / Q000378:metabolism*; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis*; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D010449:Peptide Mapping / Q000379:methods*; D012680:Sensitivity and Specificity; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D042501:Angiogenic Proteins; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins