Sociedade Brasileira de Espectrometria de Massas – BrMASS
DIFFERENTIAL PROTEOMIC ANALYSIS IN THE MYOCARDIUM
FROM PATIENTS WITH CHRONIC CHAGAS DISEASE
CARDIOMYOPATHY
Priscila C. Teixeira1,2, Axel Ducret4, Vilem Guryca4, Hanno Langen4, Andréia
Kuramoto1, Ronaldo Honorato3, Alfredo Fiorelli3, Noedir Stolf3, Jorge
Kalil1,2, Edecio Cunha-Neto1,2
[email protected]
1
Laboratory of Immunology, Heart Institute - University of São Paulo,
Division of Clinical Immunology and Allergy, School of Medicine - University
of São Paulo, 3Division of Surgery, Heart Institute - University of São Paulo,
São Paulo, Brazil. 4Molecular Medicine Laboratories, F. Hoffmann-La Roche,
Basel, Switzerland.
2
Chronic Chagas disease cardiomyopathy (CCC), caused by Trypanosoma cruzi,
shows a shorter survival than other non-inflammatory cardiomyopathies, such as
ischemic (IC) and idiopathic dilated cardiomyopathies (IDC). To gain a better
understanding of the CCC pathophysiology, we analyzed the myocardial protein and
gene expression of enzymes related to ATP production in samples from patients with
CCC, IC and IDC, and also from individuals without cardiomyopathy (N). Myocardium
homogenates were subjected to two-dimensional electrophoresis with fluorescent
labeling (DIGE) and protein identification by mass spectrometry (MALDI-ToF/ToF) to
analyze the differential protein expression profile in the myocardium of patients
afflicted by CCC, IDC and IC. We identified a total of 683 spots, corresponding to 230
distinct proteins. We observed that the protein expression profile of CCC patients is the
most distinct when compared to non-cardiomyopathic subjects. On the other hand, the
protein expression profile of IC patients is similar, at some extent, to the expression
profile of non-cardiomyopathic patients. We also found altered expression of proteins
related to apoptosis, oxidative stress, endoplasmic reticulum stress and cardiac
remodeling among CCC, IDC and IC patients when compared to non-cardiomyopathic
patients. We also showed that the myocardium of patients afflicted by CCC display
altered expression of several mitochondrial proteins associated to energy metabolism in the glycolysis, Krebs cycle, beta-oxidation, oxidative phosphorylation, and creatine
kinase complex - when compared to non-cardiomyopathic subjects. Although some of
these changes were shared with IDC samples, and, to a lesser extent, with IC samples,
Western blot analysis demonstrated that CCC samples showed the most extreme
reduction in protein expression of the creatine kinase system; the same was observed
in its enzymatic activity. Taken together, our results suggest that diminished
expression of proteins fundamental for ATP generation, increased expression of
apoptosis-associated proteins and immune system proteins in the myocardium of CCC
patients when compared to IC and IDC patients may be associated to CCC progression.
The analysis of the protein expression profile by 2D-gel and mass spectrometry has
identified groups of proteins whose expression pattern is able to discriminate the
myocardium samples by etiology. This may help to find novel peripheral biomarkers of
CCC and other cardiomyopathies, as well as in the understanding of mechanisms of
disease progression and structural/molecular alterations of the inflamed myocardium.
Supported by: FAPESP/CNPq.
3º Congresso BrMass – 12 a 15 de Dezembro de 2009