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Heart mitochondrial proteome study elucidates changes in cardiac energy metabolism and antioxidant PRDX3 in human dilated cardiomyopathy.

Roselló-Lletí E, Tarazón E, Barderas MG, Ortega A, Otero M, Molina-Navarro MM, Lago F, González-Juanatey JR, Salvador A, Portolés M, Rivera M - PLoS ONE (2014)

Bottom Line: We found significant alterations in energy metabolism, especially in molecules involved in substrate utilization (ODPA, ETFD, DLDH), energy production (ATPA), other metabolic pathways (AL4A1) and protein synthesis (EFTU), obtaining considerable and specific relationships between the alterations detected in these processes.Importantly, we observed that the antioxidant PRDX3 overexpression is associated with impaired ventricular function.PRDX3 is significantly related to LV end systolic and diastolic diameter (r = 0.73, p value<0.01; r = 0.71, p value<0.01), fractional shortening, and ejection fraction (r = -0.61, p value<0.05; and r = -0.62, p value<0.05, respectively).

View Article: PubMed Central - PubMed

Affiliation: Cardiocirculatory Unit, Health Research Institute Hospital La Fe, Valencia, Spain.

ABSTRACT

Background: Dilated cardiomyopathy (DCM) is a public health problem with no available curative treatment, and mitochondrial dysfunction plays a critical role in its development. The present study is the first to analyze the mitochondrial proteome in cardiac tissue of patients with DCM to identify potential molecular targets for its therapeutic intervention.

Methods and results: 16 left ventricular (LV) samples obtained from explanted human hearts with DCM (n = 8) and control donors (n = 8) were extracted to perform a proteomic approach to investigate the variations in mitochondrial protein expression. The proteome of the samples was analyzed by quantitative differential electrophoresis and Mass Spectrometry. These changes were validated by classical techniques and by novel and precise selected reaction monitoring analysis and RNA sequencing approach increasing the total heart samples up to 25. We found significant alterations in energy metabolism, especially in molecules involved in substrate utilization (ODPA, ETFD, DLDH), energy production (ATPA), other metabolic pathways (AL4A1) and protein synthesis (EFTU), obtaining considerable and specific relationships between the alterations detected in these processes. Importantly, we observed that the antioxidant PRDX3 overexpression is associated with impaired ventricular function. PRDX3 is significantly related to LV end systolic and diastolic diameter (r = 0.73, p value<0.01; r = 0.71, p value<0.01), fractional shortening, and ejection fraction (r = -0.61, p value<0.05; and r = -0.62, p value<0.05, respectively).

Conclusion: 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.

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Mitochondrial protein overexpression in dilated human hearts according to immunofluorescence techniques.Influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and the stress response (PRDX3). Immunofluorescence of (a) ODPA, (b) ETFD, (c) DLDH, (d) AL4A1, (e) ATPA, (f) EFTU, and (g) PRDX3 were significantly increased in patients with dilated cardiomyopathy compared with the control group. Here we show the nucleus co-stained with DAPI (blue). All of the micrographs are representative of the results obtained in four independent experiments for each group and protein studied, DCM (n = 4) and CNT (n = 4). The bar represents 100 µm. The bar graph shows the relative fluorescence intensity in dilated compared to control hearts. The data are expressed as mean ± SEM. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase. *p value<0.05, **p value<0.01.
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pone-0112971-g002: Mitochondrial protein overexpression in dilated human hearts according to immunofluorescence techniques.Influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and the stress response (PRDX3). Immunofluorescence of (a) ODPA, (b) ETFD, (c) DLDH, (d) AL4A1, (e) ATPA, (f) EFTU, and (g) PRDX3 were significantly increased in patients with dilated cardiomyopathy compared with the control group. Here we show the nucleus co-stained with DAPI (blue). All of the micrographs are representative of the results obtained in four independent experiments for each group and protein studied, DCM (n = 4) and CNT (n = 4). The bar represents 100 µm. The bar graph shows the relative fluorescence intensity in dilated compared to control hearts. The data are expressed as mean ± SEM. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase. *p value<0.05, **p value<0.01.

Mentions: 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 dilated hearts than in the CNT samples. These proteins had a diffuse cytoplasmic distribution with a significantly higher percentage of fluorescence in the dilated group (ODPA, 21%, p value<0.01 [Fig. 2a]; ETFD, 44%, p value<0.01 [Fig. 2b]; DLDH, 24%, p value<0.05 [Fig. 2c]; AL4A1, 32%, p value<0.01 [Fig. 2d]; ATPA, 18%, p value<0.01 [Fig. 2e]; EFTU, 42%, p value<0.05 [Fig. 2f]; PRDX3, 29%, p value<0.05 [Fig. 2g]).


Heart mitochondrial proteome study elucidates changes in cardiac energy metabolism and antioxidant PRDX3 in human dilated cardiomyopathy.

Roselló-Lletí E, Tarazón E, Barderas MG, Ortega A, Otero M, Molina-Navarro MM, Lago F, González-Juanatey JR, Salvador A, Portolés M, Rivera M - PLoS ONE (2014)

Mitochondrial protein overexpression in dilated human hearts according to immunofluorescence techniques.Influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and the stress response (PRDX3). Immunofluorescence of (a) ODPA, (b) ETFD, (c) DLDH, (d) AL4A1, (e) ATPA, (f) EFTU, and (g) PRDX3 were significantly increased in patients with dilated cardiomyopathy compared with the control group. Here we show the nucleus co-stained with DAPI (blue). All of the micrographs are representative of the results obtained in four independent experiments for each group and protein studied, DCM (n = 4) and CNT (n = 4). The bar represents 100 µm. The bar graph shows the relative fluorescence intensity in dilated compared to control hearts. The data are expressed as mean ± SEM. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase. *p value<0.05, **p value<0.01.
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pone-0112971-g002: Mitochondrial protein overexpression in dilated human hearts according to immunofluorescence techniques.Influence of dilated cardiomyopathy on the amount of each representative protein involved in cardiac energy metabolism (ODPA, ETFD, DLDH, AL4A1, and ATPA), protein biosynthesis (EFTU), and the stress response (PRDX3). Immunofluorescence of (a) ODPA, (b) ETFD, (c) DLDH, (d) AL4A1, (e) ATPA, (f) EFTU, and (g) PRDX3 were significantly increased in patients with dilated cardiomyopathy compared with the control group. Here we show the nucleus co-stained with DAPI (blue). All of the micrographs are representative of the results obtained in four independent experiments for each group and protein studied, DCM (n = 4) and CNT (n = 4). The bar represents 100 µm. The bar graph shows the relative fluorescence intensity in dilated compared to control hearts. The data are expressed as mean ± SEM. CNT, control; DCM, dilated cardiomyopathy; ODPA, pyruvate dehydrogenase E1 component subunit α, somatic form; ETFD, electron transfer flavoprotein-ubiquinone oxidoreductase; DLDH, dihydrolipoyl dehydrogenase; AL4A1, delta-1-pyrroline-5-carboxylate dehydrogenase; ATPA, ATP synthase subunit α; EFTU, elongation factor Tu; PRDX3, thioredoxin-dependent peroxide reductase. *p value<0.05, **p value<0.01.
Mentions: 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 dilated hearts than in the CNT samples. These proteins had a diffuse cytoplasmic distribution with a significantly higher percentage of fluorescence in the dilated group (ODPA, 21%, p value<0.01 [Fig. 2a]; ETFD, 44%, p value<0.01 [Fig. 2b]; DLDH, 24%, p value<0.05 [Fig. 2c]; AL4A1, 32%, p value<0.01 [Fig. 2d]; ATPA, 18%, p value<0.01 [Fig. 2e]; EFTU, 42%, p value<0.05 [Fig. 2f]; PRDX3, 29%, p value<0.05 [Fig. 2g]).

Bottom Line: We found significant alterations in energy metabolism, especially in molecules involved in substrate utilization (ODPA, ETFD, DLDH), energy production (ATPA), other metabolic pathways (AL4A1) and protein synthesis (EFTU), obtaining considerable and specific relationships between the alterations detected in these processes.Importantly, we observed that the antioxidant PRDX3 overexpression is associated with impaired ventricular function.PRDX3 is significantly related to LV end systolic and diastolic diameter (r = 0.73, p value<0.01; r = 0.71, p value<0.01), fractional shortening, and ejection fraction (r = -0.61, p value<0.05; and r = -0.62, p value<0.05, respectively).

View Article: PubMed Central - PubMed

Affiliation: Cardiocirculatory Unit, Health Research Institute Hospital La Fe, Valencia, Spain.

ABSTRACT

Background: Dilated cardiomyopathy (DCM) is a public health problem with no available curative treatment, and mitochondrial dysfunction plays a critical role in its development. The present study is the first to analyze the mitochondrial proteome in cardiac tissue of patients with DCM to identify potential molecular targets for its therapeutic intervention.

Methods and results: 16 left ventricular (LV) samples obtained from explanted human hearts with DCM (n = 8) and control donors (n = 8) were extracted to perform a proteomic approach to investigate the variations in mitochondrial protein expression. The proteome of the samples was analyzed by quantitative differential electrophoresis and Mass Spectrometry. These changes were validated by classical techniques and by novel and precise selected reaction monitoring analysis and RNA sequencing approach increasing the total heart samples up to 25. We found significant alterations in energy metabolism, especially in molecules involved in substrate utilization (ODPA, ETFD, DLDH), energy production (ATPA), other metabolic pathways (AL4A1) and protein synthesis (EFTU), obtaining considerable and specific relationships between the alterations detected in these processes. Importantly, we observed that the antioxidant PRDX3 overexpression is associated with impaired ventricular function. PRDX3 is significantly related to LV end systolic and diastolic diameter (r = 0.73, p value<0.01; r = 0.71, p value<0.01), fractional shortening, and ejection fraction (r = -0.61, p value<0.05; and r = -0.62, p value<0.05, respectively).

Conclusion: 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.

Show MeSH
Related in: MedlinePlus