Serum proteomic profiles in CKCS with Mitral valve disease.
BACKGROUND
Myxomatous mitral valve disease (MVD) is the most common acquired heart disease in dogs, and the Cavalier King Charles Spaniel (CKCS) is the most studied breed because of the high prevalence, early onset and hereditary component evidenced in the breed. MVD has different severity levels, and there are many practical limitations in identifying its asymptomatic stages. Proteomic techniques are valuable for studying the proteins and peptides involved in cardiovascular diseases, including the period prior to the clinical onset of the disease. The aim of this study was to identify the serum proteins that were differentially expressed in healthy CKCS and those affected by MVD in mild to severe stages. Proteomics analysis was performed using two-dimensional gel electrophoresis separation and a bioinformatics analysis for the detection of differentially expressed spots. In a comparative analysis, protein spots with a p < 0.05 (ANOVA) were considered statistically significant and were excised from the gels for analysis by MALDI-TOF-MS for protein identification.


RESULTS
Eight proteins resulted differentially expressed among the groups and significantly related to the progression of the disease. In mild affected group versus healthy dogs complement factor H isoform 2, inhibitor of carbonic anhydrase, hemopexin, dystrobrevin beta isoform X7 and CD5 molecule-like resulted to be down-regulated, whereas fibronectin type-III domain-containing protein 3A isoform X4 was up-regulated. In severe affected dogs versus healthy group complement factor H isoform 2, calpain-3 isoform X2, dystrobrevin beta isoform X7, CD5 molecule-like and l-2-hydroxyglutarate dehydrogenase resulted to be down-regulated. Complement factor H isoform 2, calpain-3 isoform X2, dystrobrevin beta isoform X7, CD5 molecule-like and hydroxyglutarate dehydrogenase were found to be down-regulated in mild affected group versus healthy dogs. All of these proteins except complement factor H followed a decreasing trend according to the progression of the pathology.


CONCLUSIONS
The differential expression of serum proteins demonstrates the possibility these might be valuable for the detection and monitoring of the disease. Further longitudinal studies are required to determine whether differential protein expression occurs sufficiently early in the progression of the disease and with sufficient predictive value to allow proteomics analysis to be used as an early detection and on-line diagnostic tool.
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Chiara
C
Locatelli
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Cristian
C
Piras
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Giulia
G
Riscazzi
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Isabella
I
Alloggio
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Ilaria
I
Spalla
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Alessio
A
Soggiu
Fondazione Santa Lucia -IRCCS, Rome, Italy.
Viviana
V
Greco
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.
Luigi
L
Bonizzi
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy.|Istituto Sperimentale Italiano L. Spallanzani, Milan, Italy.
Paola
P
Roncada
DIMEVET, Department of Veterinary Medicine, University of Milan, Milan, Italy. paola.brambilla@unimi.it.
Paola G
PG
Brambilla
D000818:Animals; D001798:Blood Proteins / Q000032:analysis; D001947:Breeding; D016022:Case-Control Studies; D004283:Dog Diseases / Q000097:blood* / Q000175:diagnosis; D004285:Dogs; D005260:Female; D006349:Heart Valve Diseases / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D008297:Male; D008943:Mitral Valve / Q000473:pathology; D020543:Proteome; D040901:Proteomics
D001798:Blood Proteins; D020543:Proteome
Cavalier King Charles Spaniel; Dog; Mitral valve disease; Proteomic analysis