Abstract 11515: Characterization of Heart Mitochondrial Proteome Elucidates Significant Changes in Cardiac Energy Metabolism and Antioxidant Prdx3 in Human Dilated Cardiomyopathy
Introduction: Dilated cardiomyopathy (DCM) is a public health problem with no available curative treatment, and mitochondrial dysfunction plays a critical role in its development.
Methods: LV tissue samples were obtained from 17 patients with DCM. Eight non-diseased donor hearts were used as CNT samples. The protein expressions of purified heart mitochondria from 8 DCM patients and 8 CNT were compared using two-dimensional differential gel electrophoresis (2DDIGE).
Results: We encountered 16 significantly upregulated and 5 downregulated spots in DCM hearts. A selection of representative proteins of each mitochondrial function, localization was validated using different techniques that compared its levels among DCM patients (n = 17) and CNT (n = 8). Therefore, using western blot techniques, we determined the levels of some relevant proteins involved in metabolism, including pyruvate dehydrogenase E1 component subunit alpha, somatic form (ODPA); ETFD; DLDH; delta-1-pyrroline-5-carboxylate dehydrogenase (AL4A1); ATPA; protein synthesis EFTU; and stress response PRDX3. We found significant relationships between PRDX3 levels and LV function. The results obtained showed that PRDX3 is significantly correlated with fractional shortening (p<0.01), and ejection fraction (p<0.01). The immunofluorescence study findings were consistent with the increased levels observed by western blotting and proteomic analysis, showing that the intensity of all validated proteins was higher in the DCM hearts than in the CNT. Immunocytochemistry confirmed the previous results and the localization or distribution of these mitochondrial proteins. To validate the previous analyses and to evaluate the possible relationship between the altered proteins involved in the different mitochondrial processes, these molecules were monitored by SRM. The mRNA differences between DCM patients and CNT were determined by RNA sequencing analysis.
Conclusions: This work could be a pivotal study to gain more knowledge on the cellular mechanisms related to the pathophysiology of this disease and may lead to the development of etiology specific heart failure therapies. We suggest new molecular targets for therapeutic interventions, something that up to now has been lacking.
Author Disclosures: M. Rivera: None. E. Rosello: None. M. Barderas: None. E. Tarazon: None. M. Molina: None. A. Ortega: None. L. Rodriguez Padial: None. M. Portoles: None.
- © 2014 by American Heart Association, Inc.