Bio

Bio


PhD degree in Physiology (University of Sao Paulo) and postdoc in Biochemistry (Stanford University). The main focus of my research is to understand the role of mitochondria as intracellular node (not just the powerhouse of the cell) and their impact on life/death decision.

Academic Appointments


Patents


  • Julio Ferreira. "United StatesAntagonists of Mitofusion I and Beta II PKC Association for Treating Heart Failure"

Publications

All Publications


  • Mitochondrially-targeted treatment strategies. Molecular aspects of medicine Bozi, L. H., Campos, J. C., Zambelli, V. O., Ferreira, N. D., Ferreira, J. C. 2019: 100836

    Abstract

    Disruption of mitochondrial function is a common feature of inherited mitochondrial diseases (mitochondriopathies) and many other infectious and non-infectious diseases including viral, bacterial and protozoan infections, inflammatory and chronic pain, neurodegeneration, diabetes, obesity and cardiovascular diseases. Mitochondria therefore become an attractive target for developing new therapies. In this review we describe critical mechanisms involved in the maintenance of mitochondrial functionality and discuss strategies used to identify and validate mitochondrial targets in different diseases. We also highlight the most recent preclinical and clinical findings using molecules targeting mitochondrial bioenergetics, morphology, number, content and detoxification systems in common pathologies.

    View details for DOI 10.1016/j.mam.2019.100836

    View details for PubMedID 31866004

  • Mitochondrial Bioenergetics and Quality Control Mechanisms in Health and Disease OXIDATIVE MEDICINE AND CELLULAR LONGEVITY Ferreira, J. B., Mori, M. A., Gross, E. R. 2019; 2019
  • A selective inhibitor of mitofusin 1-betaIIPKC association improves heart failure outcome in rats. Nature communications Ferreira, J. C., Campos, J. C., Qvit, N., Qi, X., Bozi, L. H., Bechara, L. R., Lima, V. M., Queliconi, B. B., Disatnik, M., Dourado, P. M., Kowaltowski, A. J., Mochly-Rosen, D. 2019; 10 (1): 329

    Abstract

    We previously demonstrated that beta II protein kinase C (betaIIPKC) activity is elevated in failing hearts and contributes to this pathology. Here we report that betaIIPKC accumulates on the mitochondrial outer membrane and phosphorylates mitofusin 1 (Mfn1) at serine 86. Mfn1 phosphorylation results in partial loss of its GTPase activity and in a buildup of fragmented and dysfunctional mitochondria in heart failure. betaIIPKC siRNA or a betaIIPKC inhibitor mitigates mitochondrial fragmentation and cell death. We confirm that Mfn1-betaIIPKC interaction alone is critical in inhibiting mitochondrial function and cardiac myocyte viability using SAMbetaA, a rationally-designed peptide that selectively antagonizes Mfn1-betaIIPKC association. SAMbetaA treatment protects cultured neonatal and adult cardiac myocytes, but not Mfn1 knockout cells, from stress-induced death. Importantly, SAMbetaA treatment re-establishes mitochondrial morphology and function and improves cardiac contractility in rats with heart failure, suggesting that SAMbetaA may be a potential treatment for patients with heart failure.

    View details for PubMedID 30659190

  • A selective inhibitor of mitofusin 1-beta IIPKC association improves heart failure outcome in rats NATURE COMMUNICATIONS Ferreira, J. B., Campos, J. C., Qvit, N., Qi, X., Bozi, L. M., Bechara, L. G., Lima, V. M., Queliconi, B. B., Disatnik, M., Dourado, P. M., Kowaltowski, A. J., Mochly-Rosen, D. 2019; 10
  • ALDH2 and Cardiovascular Disease. Advances in experimental medicine and biology Chen, C., Ferreira, J. C., Mochly-Rosen, D. 2019; 1193: 53?67

    Abstract

    Aldehyde dehydrogenase 2 (ALDH2) is a non-cytochrome P450 mitochondrial aldehyde oxidizing enzyme. It is best known for its role in the metabolism of acetaldehyde, a common metabolite from alcohol drinking. More evidences have been accumulated in recent years to indicate a greater role of ALDH2 in the metabolism of other endogenous and exogenous aldehydes, especially lipid peroxidation-derived reactive aldehyde under oxidative stress. Many cardiovascular diseases are associated with oxidative stress and mitochondria dysfunction. Considering that an estimated 560million East Asians carry a common ALDH2 deficient variant which causes the well-known alcohol flushing syndrome due to acetaldehyde accumulation, the importance of understanding the role of ALDH2 in these diseases should be highlighted. There are several unfavorable cardiovascular conditions that are associated with ALDH2 deficiency. This chapter reviews the function of ALDH2 in various pathological conditions of the heart in relation to aldehyde toxicity. It also highlights the importance and clinical implications of interaction between ALDH2 deficiency and alcohol drinking on cardiovascular disease among the East Asians.

    View details for DOI 10.1007/978-981-13-6260-6_3

    View details for PubMedID 31368097

  • Mitochondrial Bioenergetics and Quality Control Mechanisms in Health and Disease. Oxidative medicine and cellular longevity Ferreira, J. C., Mori, M. A., Gross, E. R. 2019; 2019: 5406751

    View details for PubMedID 30805083

  • Alcohol consumption and vascular disease: other points to consider. Lancet (London, England) Chen, C. H., Ferreira, J. C., Mochly-Rosen, D., Gross, E. R. 2019; 394 (10209): 1617?18

    View details for DOI 10.1016/S0140-6736(19)31880-X

    View details for PubMedID 31690444

  • Targeting mitochondrial dysfunction and oxidative stress in heart failure: Challenges and opportunities FREE RADICAL BIOLOGY AND MEDICINE Kiyuna, L., Prestes e Albuquerque, R., Chen, C., Mochly-Rosen, D., Batista Ferreira, J. 2018; 129: 155?68

    Abstract

    Mitochondrial dysfunction characterized by impaired bioenergetics, oxidative stress and aldehydic load is a hallmark of heart failure. Recently, different research groups have provided evidence that selective activation of mitochondrial detoxifying systems that counteract excessive accumulation of ROS, RNS and reactive aldehydes is sufficient to stop cardiac degeneration upon chronic stress, such as heart failure. Therefore, pharmacological and non-pharmacological approaches targeting mitochondria detoxification may play a critical role in the prevention or treatment of heart failure. In this review we discuss the most recent findings on the central role of mitochondrial dysfunction, oxidative stress and aldehydic load in heart failure, highlighting the most recent preclinical and clinical studies using mitochondria-targeted molecules and exercise training as effective tools against heart failure.

    View details for PubMedID 30227272

  • 4-HNE-mediated post-translational modulation of DICER in heart failure Kiyuna, L., MacRae, I. J., Chen, C., Mochly-Rosen, D., Ferreira, J. ELSEVIER SCIENCE INC. 2018: S29
  • Cardioprotection induced by a brief exposure to acetaldehyde: role of aldehyde dehydrogenase 2 CARDIOVASCULAR RESEARCH Ueta, C., Campos, J., Prestes e Albuquerque, R., Lima, V., Disatnik, M., Sanchez, A., Chen, C., Gennari de Medeiros, M., Yang, W., Mochly-Rosen, D., Batista Ferreira, J. 2018; 114 (7): 1006?15

    Abstract

    We previously demonstrated that acute ethanol administration protects the heart from ischaemia/reperfusion (I/R) injury thorough activation of aldehyde dehydrogenase 2 (ALDH2). Here, we characterized the role of acetaldehyde, an intermediate product from ethanol metabolism, and its metabolizing enzyme, ALDH2, in an ex vivo model of cardiac I/R injury.We used a combination of homozygous knock-in mice (ALDH2*2), carrying the human inactivating point mutation ALDH2 (E487K), and a direct activator of ALDH2, Alda-1, to investigate the cardiac effect of acetaldehyde. The ALDH2*2 mice have impaired acetaldehyde clearance, recapitulating the human phenotype. Yet, we found a similar infarct size in wild type (WT) and ALDH2*2 mice. Similar to ethanol-induced preconditioning, pre-treatment with 50 ?M acetaldehyde increased ALDH2 activity and reduced cardiac injury in hearts of WT mice without affecting cardiac acetaldehyde levels. However, acetaldehyde pre-treatment of hearts of ALDH2*2 mice resulted in a three-fold increase in cardiac acetaldehyde levels and exacerbated I/R injury. Therefore, exogenous acetaldehyde appears to have a bimodal effect in I/R, depending on the ALDH2 genotype. Further supporting an ALDH2 role in cardiac preconditioning, pharmacological ALDH2 inhibition abolished ethanol-induced cardioprotection in hearts of WT mice, whereas a selective activator, Alda-1, protected ALDH2*2 against ethanol-induced cardiotoxicity. Finally, either genetic or pharmacological inhibition of ALDH2 mitigated ischaemic preconditioning.Taken together, our findings suggest that low levels of acetaldehyde are cardioprotective whereas high levels are damaging in an ex vivo model of I/R injury and that ALDH2 is a major, but not the only, regulator of cardiac acetaldehyde levels and protection from I/R.

    View details for PubMedID 29579152

    View details for PubMedCentralID PMC5967552

  • Disruption of mitochondrial quality control in peripheral artery disease: New therapeutic opportunities PHARMACOLOGICAL RESEARCH Ueta, C. B., Gomes, K. S., Ribeiro, M. A., Mochly-Rosen, D., Ferreira, J. C. 2017; 115: 96-106

    Abstract

    Peripheral artery disease (PAD) is a multifactorial disease initially triggered by reduced blood supply to the lower extremities due to atherosclerotic obstructions. It is considered a major public health problem worldwide, affecting over 200 million people. Management of PAD includes smoking cessation, exercise, statin therapy, antiplatelet therapy, antihypertensive therapy and surgical intervention. Although these pharmacological and non-pharmacological interventions usually increases blood flow to the ischemic limb, morbidity and mortality associated with PAD continue to increase. This scenario raises new fundamental questions regarding the contribution of intrinsic metabolic changes in the distal affected skeletal muscle to the progression of PAD. Recent evidence suggests that disruption of skeletal muscle mitochondrial quality control triggered by intermittent ischemia-reperfusion injury is associated with increased morbidity in individuals with PAD. The mitochondrial quality control machinery relies on surveillance systems that help maintaining mitochondrial homeostasis upon stress. In this review, we describe some of the most critical mechanisms responsible for the impaired skeletal muscle mitochondrial quality control in PAD. We also discuss recent findings on the central role of mitochondrial bioenergetics and quality control mechanisms including mitochondrial fusion-fission balance, turnover, oxidative stress and aldehyde metabolism in the pathophysiology of PAD, and highlight their potential as therapeutic targets.

    View details for DOI 10.1016/j.phrs.2016.11.016

    View details for PubMedID 27876411

  • Exercise reestablishes autophagic flux and mitochondrial quality control in heart failure AUTOPHAGY Campos, J. C., Queliconi, B. B., Bozi, L. M., Bechara, L. G., Dourado, P. M., Andres, A. M., Jannig, P. R., Gomes, K. S., Zambelli, V. O., Rocha-Resende, C., Guatimosim, S., Brum, P. C., Mochly-Rosen, D., Gottlieb, R. A., Kowaltowski, A. J., Ferreira, J. B. 2017; 13 (8): 1304?17

    Abstract

    We previously reported that facilitating the clearance of damaged mitochondria through macroautophagy/autophagy protects against acute myocardial infarction. Here we characterize the impact of exercise, a safe strategy against cardiovascular disease, on cardiac autophagy and its contribution to mitochondrial quality control, bioenergetics and oxidative damage in a post-myocardial infarction-induced heart failure animal model. We found that failing hearts displayed reduced autophagic flux depicted by accumulation of autophagy-related markers and loss of responsiveness to chloroquine treatment at 4 and 12wk after myocardial infarction. These changes were accompanied by accumulation of fragmented mitochondria with reduced O2 consumption, elevated H2O2 release and increased Ca2+-induced mitochondrial permeability transition pore opening. Of interest, disruption of autophagic flux was sufficient to decrease cardiac mitochondrial function in sham-treated animals and increase cardiomyocyte toxicity upon mitochondrial stress. Importantly, 8wk of exercise training, starting 4wk after myocardial infarction at a time when autophagy and mitochondrial oxidative capacity were already impaired, improved cardiac autophagic flux. These changes were followed by reduced mitochondrial number:size ratio, increased mitochondrial bioenergetics and better cardiac function. Moreover, exercise training increased cardiac mitochondrial number, size and oxidative capacity without affecting autophagic flux in sham-treated animals. Further supporting an autophagy mechanism for exercise-induced improvements of mitochondrial bioenergetics in heart failure, acute in vivo inhibition of autophagic flux was sufficient to mitigate the increased mitochondrial oxidative capacity triggered by exercise in failing hearts. Collectively, our findings uncover the potential contribution of exercise in restoring cardiac autophagy flux in heart failure, which is associated with better mitochondrial quality control, bioenergetics and cardiac function.

    View details for PubMedID 28598232

  • Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) Protein-Protein Interaction Inhibitor Reveals a Non-catalytic Role for GAPDH Oligomerization in Cell Death JOURNAL OF BIOLOGICAL CHEMISTRY Qvit, N., Joshi, A. U., Cunningham, A. D., Ferreira, J. C., Mochly-Rosen, D. 2016; 291 (26): 13608-13621

    Abstract

    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an important glycolytic enzyme, has a non-catalytic (thus a non-canonical) role in inducing mitochondrial elimination under oxidative stress. We recently demonstrated that phosphorylation of GAPDH by delta protein kinase C (PKC) inhibits this GAPDH-dependent mitochondrial elimination. deltaPKC phosphorylation of GAPDH correlates with increased cell injury following oxidative stress, suggesting that inhibiting GAPDH phosphorylation should decrease cell injury. Using rational design, we identified pseudoGAPDH peptide, an inhibitor of deltaPKC-mediated GAPDH phosphorylation that does not inhibit the phosphorylation of other deltaPKC substrates. Unexpectedly, pseudoGAPDH decreased mitochondrial elimination and increased cardiac damage in an animal model of heart attack. Either treatment with pseudoGAPDH or direct phosphorylation of GAPDH by deltaPKC decreased GAPDH tetramerization, which corresponded to reduced GAPDH glycolytic activity in vitro and ex vivo. Taken together, our study identified the potential mechanism by which oxidative stress inhibits the protective GAPDH-mediated elimination of damaged mitochondria. Our study also identified a pharmacological tool, pseudoGAPDH peptide, with interesting properties. pseudoGAPDH peptide is an inhibitor of the interaction between deltaPKC and GAPDH, and of the resulting phosphorylation of GAPDH by deltaPKC. pseudoGAPDH peptide is also an inhibitor of GAPDH oligomerization and thus an inhibitor of GAPDH glycolytic activity. Finally, we found that pseudoGAPDH peptide is an inhibitor of the elimination of damaged mitochondria. We discuss how this unique property of increasing cell damage following oxidative stress suggests a potential use for pseudoGAPDH peptide-based therapy.

    View details for DOI 10.1074/jbc.M115.711630

    View details for PubMedID 27129213

  • New therapeutics to modulate mitochondrial dynamics and mitophagy in cardiac diseases. Journal of molecular medicine (Berlin, Germany) Disatnik, M., Hwang, S., Ferreira, J. C., Mochly-Rosen, D. 2015; 93 (3): 279-287

    Abstract

    The processes that control the number and shape of the mitochondria (mitochondrial dynamics) and the removal of damaged mitochondria (mitophagy) have been the subject of intense research. Recent work indicates that these processes may contribute to the pathology associated with cardiac diseases. This review describes some of the key proteins that regulate these processes and their potential as therapeutic targets for cardiac diseases.

    View details for DOI 10.1007/s00109-015-1256-4

    View details for PubMedID 25652199

  • Aldehydic load and aldehyde dehydrogenase 2 profile during the progression of post-myocardial infarction cardiomyopathy: Benefits of Alda-1 INTERNATIONAL JOURNAL OF CARDIOLOGY Gomes, K. M., Bechara, L. R., Lima, V. M., Ribeiro, M. A., Campos, J. C., Dourado, P. M., Kowaltowski, A. J., Mochly-Rosen, D., Ferreira, J. C. 2015; 179: 129-138

    Abstract

    We previously demonstrated that reducing cardiac aldehydic load by aldehyde dehydrogenase 2 (ALDH2), a mitochondrial enzyme responsible for metabolizing the major lipid peroxidation product, protects against acute ischemia/reperfusion injury and chronic heart failure. However, time-dependent changes in ALDH2 profile, aldehydic load and mitochondrial bioenergetics during progression of post-myocardial infarction (post-MI) cardiomyopathy are unknown and should be established to determine the optimal time window for drug treatment.Here we characterized cardiac ALDH2 activity and expression, lipid peroxidation, 4-hydroxy-2-nonenal (4-HNE) adduct formation, glutathione pool and mitochondrial energy metabolism and H?O? release during the 4 weeks after permanent left anterior descending (LAD) coronary artery occlusion in rats.We observed a sustained disruption of cardiac mitochondrial function during the progression of post-MI cardiomyopathy, characterized by >50% reduced mitochondrial respiratory control ratios and up to 2 fold increase in H?O? release. Mitochondrial dysfunction was accompanied by accumulation of cardiac and circulating lipid peroxides and 4-HNE protein adducts and down-regulation of electron transport chain complexes I and V. Moreover, increased aldehydic load was associated with a 90% reduction in cardiac ALDH2 activity and increased glutathione pool. Further supporting an ALDH2 mechanism, sustained Alda-1 treatment (starting 24h after permanent LAD occlusion surgery) prevented aldehydic overload, mitochondrial dysfunction and improved ventricular function in post-MI cardiomyopathy rats.Taken together, our findings demonstrate a disrupted mitochondrial metabolism along with an insufficient cardiac ALDH2-mediated aldehyde clearance during the progression of ventricular dysfunction, suggesting a potential therapeutic value of ALDH2 activators during the progression of post-myocardial infarction cardiomyopathy.

    View details for DOI 10.1016/j.ijcard.2014.10.140

    View details for PubMedID 25464432

    View details for PubMedCentralID PMC4405147

  • A personalized medicine approach for asian americans with the aldehyde dehydrogenase 2*2 variant. Annual review of pharmacology and toxicology Gross, E. R., Zambelli, V. O., Small, B. A., Ferreira, J. C., Chen, C., Mochly-Rosen, D. 2015; 55: 107-127

    Abstract

    Asian Americans are one of the fastest-growing populations in the United States. A relatively large subset of this population carries a unique loss-of-function point mutation in aldehyde dehydrogenase 2 (ALDH2), ALDH2*2. Found in approximately 560 million people of East Asian descent, ALDH2*2 reduces enzymatic activity by approximately 60% to 80% in heterozygotes. Furthermore, this variant is associated with a higher risk for several diseases affecting many organ systems, including a particularly high incidence relative to the general population of esophageal cancer, myocardial infarction, and osteoporosis. In this review, we discuss the pathophysiology associated with the ALDH2*2 variant, describe why this variant needs to be considered when selecting drug treatments, and suggest a personalized medicine approach for Asian American carriers of this variant. We also discuss future clinical and translational perspectives regarding ALDH2*2 research.

    View details for DOI 10.1146/annurev-pharmtox-010814-124915

    View details for PubMedID 25292432

  • Aldehyde dehydrogenase 2 activation in heart failure restores mitochondrial function and improves ventricular function and remodelling CARDIOVASCULAR RESEARCH Gomes, K. M., Campos, J. C., Bechara, L. R., Queliconi, B., Lima, V. M., Disatnik, M., Magno, P., Chen, C., Brum, P. C., Kowaltowski, A. J., Mochly-Rosen, D., Ferreira, J. C. 2014; 103 (4): 498-508

    Abstract

    We previously demonstrated that pharmacological activation of mitochondrial aldehyde dehydrogenase 2 (ALDH2) protects the heart against acute ischaemia/reperfusion injury. Here, we determined the benefits of chronic activation of ALDH2 on the progression of heart failure (HF) using a post-myocardial infarction model.We showed that a 6-week treatment of myocardial infarction-induced HF rats with a selective ALDH2 activator (Alda-1), starting 4 weeks after myocardial infarction at a time when ventricular remodelling and cardiac dysfunction were present, improved cardiomyocyte shortening, cardiac function, left ventricular compliance and diastolic function under basal conditions, and after isoproterenol stimulation. Importantly, sustained Alda-1 treatment showed no toxicity and promoted a cardiac anti-remodelling effect by suppressing myocardial hypertrophy and fibrosis. Moreover, accumulation of 4-hydroxynonenal (4-HNE)-protein adducts and protein carbonyls seen in HF was not observed in Alda-1-treated rats, suggesting that increasing the activity of ALDH2 contributes to the reduction of aldehydic load in failing hearts. ALDH2 activation was associated with improved mitochondrial function, including elevated mitochondrial respiratory control ratios and reduced H2O2 release. Importantly, selective ALDH2 activation decreased mitochondrial Ca(2+)-induced permeability transition and cytochrome c release in failing hearts. Further supporting a mitochondrial mechanism for ALDH2, Alda-1 treatment preserved mitochondrial function upon in vitro aldehydic load.Selective activation of mitochondrial ALDH2 is sufficient to improve the HF outcome by reducing the toxic effects of aldehydic overload on mitochondrial bioenergetics and reactive oxygen species generation, suggesting that ALDH2 activators, such as Alda-1, have a potential therapeutic value for treating HF patients.

    View details for DOI 10.1093/cvr/cvu125

    View details for PubMedID 24817685

  • Peripheral Sensitization Increases Opioid Receptor Expression and Activation by Crotalphine in Rats PLOS ONE Zambelli, V. O., de Oliveira Fernandes, A. C., Gutierrez, V. P., Batista Ferreira, J. C., Parada, C. A., Mochly-Rosen, D., Cury, Y. 2014; 9 (3)

    Abstract

    Inflammation enhances the peripheral analgesic efficacy of opioid drugs, but the mechanisms involved in this phenomenon have not been fully elucidated. Crotalphine (CRP), a peptide that was first isolated from South American rattlesnake C.d. terrificus venom, induces a potent and long-lasting anti-nociceptive effect that is mediated by the activation of peripheral opioid receptors. Because the high efficacy of CRP is only observed in the presence of inflammation, we aimed to elucidate the mechanisms involved in the CRP anti-nociceptive effect induced by inflammation. Using real-time RT-PCR, western blot analysis and ELISA assays, we demonstrate that the intraplantar injection of prostaglandin E2 (PGE2) increases the mRNA and protein levels of the - and ?-opioid receptors in the dorsal root ganglia (DRG) and paw tissue of rats within 3 h of the injection. Using conformation state-sensitive antibodies that recognize activated opioid receptors, we show that PGE2, alone does not increase the activation of these opioid receptors but that in the presence of PGE2, the activation of specific opioid receptors by CRP and selective - and ?-opioid receptor agonists (positive controls) increases. Furthermore, PGE2 down-regulated the expression and activation of the ?-opioid receptor. CRP increased the level of activated mitogen-activated protein kinases in cultured DRG neurons, and this increase was dependent on the activation of protein kinase C?. This CRP effect was much more prominent when the cells were pretreated with PGE2. These results indicate that the expression and activation of peripheral opioid receptors by opioid-like drugs can be up- or down-regulated in the presence of an acute injury and that acute tissue injury enhances the efficacy of peripheral opioids.

    View details for DOI 10.1371/journal.pone.0090576

    View details for PubMedID 24594607

  • TARGETING ALDEHYDE DEHYDROGENASE 2: NEW THERAPEUTIC OPPORTUNITIES PHYSIOLOGICAL REVIEWS Chen, C., Batista Ferreira, J. C., Gross, E. R., Mochly-Rosen, D. 2014; 94 (1): 1-34

    Abstract

    A family of detoxifying enzymes called aldehyde dehydrogenases (ALDHs) has been a subject of recent interest, as its role in detoxifying aldehydes that accumulate through metabolism and to which we are exposed from the environment has been elucidated. Although the human genome has 19 ALDH genes, one ALDH emerges as a particularly important enzyme in a variety of human pathologies. This ALDH, ALDH2, is located in the mitochondrial matrix with much known about its role in ethanol metabolism. Less known is a new body of research to be discussed in this review, suggesting that ALDH2 dysfunction may contribute to a variety of human diseases including cardiovascular diseases, diabetes, neurodegenerative diseases, stroke, and cancer. Recent studies suggest that ALDH2 dysfunction is also associated with Fanconi anemia, pain, osteoporosis, and the process of aging. Furthermore, an ALDH2 inactivating mutation (termed ALDH2*2) is the most common single point mutation in humans, and epidemiological studies suggest a correlation between this inactivating mutation and increased propensity for common human pathologies. These data together with studies in animal models and the use of new pharmacological tools that activate ALDH2 depict a new picture related to ALDH2 as a critical health-promoting enzyme.

    View details for DOI 10.1152/physrev.00017.2013

    View details for Web of Science ID 000329194000001

    View details for PubMedID 24382882

  • Protein-Protein Inhibitor Peptide of Drp1/Fis1 Interaction Protects the Heart Against Ischemia/Reperfusion Injury Disatnik, M., Ferreira, J. C., Campos, J., Gomes, K., Dourado, P. M., Qi, X., Mochly-Rosen, D. LIPPINCOTT WILLIAMS & WILKINS. 2013
  • Acute Inhibition of Excessive Mitochondrial Fission After Myocardial Infarction Prevents Long-term Cardiac Dysfunction JOURNAL OF THE AMERICAN HEART ASSOCIATION Disatnik, M., Ferreira, J. C., Campos, J. C., Gomes, K. S., Dourado, P. M., Qi, X., Mochly-Rosen, D. 2013; 2 (5)

    Abstract

    Ischemia and reperfusion (IR) injury remains a major cause of morbidity and mortality and multiple molecular and cellular pathways have been implicated in this injury. We determined whether acute inhibition of excessive mitochondrial fission at the onset of reperfusion improves mitochondrial dysfunction and cardiac contractility postmyocardial infarction in rats.We used a selective inhibitor of the fission machinery, P110, which we have recently designed. P110 treatment inhibited the interaction of fission proteins Fis1/Drp1, decreased mitochondrial fission, and improved bioenergetics in three different rat models of IR, including primary cardiomyocytes, ex vivo heart model, and an in vivo myocardial infarction model. Drp1 transiently bound to the mitochondria following IR injury and P110 treatment blocked this Drp1 mitochondrial association. Compared with control treatment, P110 (1 ?mol/L) decreased infarct size by 28 2% and increased adenosine triphosphate levels by 70+1% after IR relative to control IR in the ex vivo model. Intraperitoneal injection of P110 (0.5 mg/kg) at the onset of reperfusion in an in vivo model resulted in improved mitochondrial oxygen consumption by 68% when measured 3 weeks after ischemic injury, improved cardiac fractional shortening by 35%, reduced mitochondrial H2O2 uncoupling state by 70%, and improved overall mitochondrial functions.Together, we show that excessive mitochondrial fission at reperfusion contributes to long-term cardiac dysfunction in rats and that acute inhibition of excessive mitochondrial fission at the onset of reperfusion is sufficient to result in long-term benefits as evidenced by inhibiting cardiac dysfunction 3 weeks after acute myocardial infarction.

    View details for DOI 10.1161/JAHA.113.000461

    View details for PubMedID 24103571

  • Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Phosphorylation by Protein Kinase C delta (PKC delta) Inhibits Mitochondria Elimination by Lysosomal-like Structures following Ischemia and Reoxygenation-induced Injury JOURNAL OF BIOLOGICAL CHEMISTRY Yogalingam, G., Hwang, S., Ferreira, J. C., Mochly-Rosen, D. 2013; 288 (26): 18947-18960

    Abstract

    After cardiac ischemia and reperfusion or reoxygenation (I/R), damaged mitochondria propagate tissue injury by promoting cell death. One possible mechanism to protect from I/R-induced injury is the elimination of damaged mitochondria by mitophagy. Here we identify new molecular events that lead to mitophagy using a cell culture model and whole hearts subjected to I/R. We found that I/R induces glyceraldehyde-3-phosphate dehydrogenase (GAPDH) association with mitochondria and promotes direct uptake of damaged mitochondria into multi-organellar lysosomal-like (LL) structures for elimination independently of the macroautophagy pathway. We also found that protein kinase C delta (?PKC) inhibits GAPDH-driven mitophagy by phosphorylating mitochondrial-associated GAPDH at threonine 246 following I/R. Phosphorylated GAPDH promotes the accumulation of mitochondria at the periphery of LL structures, which coincides with increased mitochondrial permeability. Either inhibition of ?PKC or expression of a phosphorylation-defective GAPDH mutant during I/R promotes a reduction in mitochondrial mass and apoptosis, thus indicating rescued mitophagy. Taken together, we identified a GAPDH/?PKC signaling switch, which is activated during oxidative stress to regulate the balance between cell survival by mitophagy and cell death due to accumulation of damaged mitochondria.

    View details for DOI 10.1074/jbc.M113.466870

    View details for PubMedID 23653351

  • In vivo measurement of aldehyde dehydrogenase-2 activity in rat liver ethanol model using dynamic MRSI of hyperpolarized [1-(13) C]pyruvate. NMR in biomedicine Josan, S., Xu, T., Yen, Y., Hurd, R., Ferreira, J., Chen, C., Mochly-Rosen, D., Pfefferbaum, A., Mayer, D., Spielman, D. 2013; 26 (6): 607-612

    Abstract

    To date, measurements of the activity of aldehyde dehydrogenase-2 (ALDH2), a critical mitochondrial enzyme for the elimination of certain cytotoxic aldehydes in the body and a promising target for drug development, have been largely limited to in vitro methods. Recent advancements in MRS of hyperpolarized (13) C-labeled substrates have provided a method to detect and image?in vivo metabolic pathways with signal-to-noise ratio gains greater than 10 000-fold over conventional MRS techniques. However aldehydes, because of their toxicity and short T1 relaxation times, are generally poor targets for such (13) C-labeled studies. In this work, we show that dynamic MRSI of hyperpolarized [1-(13) C]pyruvate and its conversion to [1-(13) C]lactate can provide an indirect in vivo measurement of ALDH2 activity via the concentration of NADH (nicotinamide adenine dinucleotide, reduced form), a co-factor common to both the reduction of pyruvate to lactate and the oxidation of acetaldehyde to acetate. Results from a rat liver ethanol model (n?=?9) show that changes in (13) C-lactate labeling following the bolus injection of hyperpolarized pyruvate are highly correlated with changes in ALDH2 activity (R(2) ?=?0.76). Copyright 2012 John Wiley & Sons, Ltd.

    View details for DOI 10.1002/nbm.2897

    View details for PubMedID 23225495

    View details for PubMedCentralID PMC3634870

  • Exercise Training Restores Cardiac Protein Quality Control in Heart Failure PLOS ONE Campos, J. C., Queliconi, B. B., Dourado, P. M., Cunha, T. F., Zambelli, V. O., Bechara, L. R., Kowaltowski, A. J., Brum, P. C., Mochly-Rosen, D., Ferreira, J. C. 2012; 7 (12)

    Abstract

    Exercise training is a well-known coadjuvant in heart failure treatment; however, the molecular mechanisms underlying its beneficial effects remain elusive. Despite the primary cause, heart failure is often preceded by two distinct phenomena: mitochondria dysfunction and cytosolic protein quality control disruption. The objective of the study was to determine the contribution of exercise training in regulating cardiac mitochondria metabolism and cytosolic protein quality control in a post-myocardial infarction-induced heart failure (MI-HF) animal model. Our data demonstrated that isolated cardiac mitochondria from MI-HF rats displayed decreased oxygen consumption, reduced maximum calcium uptake and elevated H?O? release. These changes were accompanied by exacerbated cardiac oxidative stress and proteasomal insufficiency. Declined proteasomal activity contributes to cardiac protein quality control disruption in our MI-HF model. Using cultured neonatal cardiomyocytes, we showed that either antimycin A or H?O? resulted in inactivation of proteasomal peptidase activity, accumulation of oxidized proteins and cell death, recapitulating our in vivo model. Of interest, eight weeks of exercise training improved cardiac function, peak oxygen uptake and exercise tolerance in MI-HF rats. Moreover, exercise training restored mitochondrial oxygen consumption, increased Ca?-induced permeability transition and reduced H?O? release in MI-HF rats. These changes were followed by reduced oxidative stress and better cardiac protein quality control. Taken together, our findings uncover the potential contribution of mitochondrial dysfunction and cytosolic protein quality control disruption to heart failure and highlight the positive effects of exercise training in re-establishing cardiac mitochondrial physiology and protein quality control, reinforcing the importance of this intervention as a non-pharmacological tool for heart failure therapy.

    View details for DOI 10.1371/journal.pone.0052764

    View details for Web of Science ID 000312829100078

    View details for PubMedID 23300764

    View details for PubMedCentralID PMC3531365

  • Identification of epsilon PKC Targets During Cardiac Ischemic Injury CIRCULATION JOURNAL Budas, G., Costa, H. M., Batista Ferreira, J. C., da Silva Ferreira, A. T., Perales, J., Krieger, J. E., Mochly-Rosen, D., Schechtman, D. 2012; 76 (6): 1476-1485

    Abstract

    Epsilon-protein kinase C (?PKC) protects the heart from ischemic injury. However, the mechanism(s) of ?PKC cardioprotection is still unclear. Identification of the ?PKC targets may aid in elucidating the ?PKC-mediated cardioprotective mechanisms. Previous studies, using ?PKC transgenic mice and difference in gel electrophoresis, identified proteins involved in glucose metabolism, the expression of which was modified by ?PKC. Those studies were accompanied by metabolomic analysis, suggesting that increased glucose oxidation may be responsible for the cardioprotective effect of ?PKC. Whether these ?PKC-mediated alterations were because of differences in protein expression or phosphorylation was not determined.In the present study, we used an ?PKC -specific activator peptide, ??RACK, combined with phosphoproteomics, to find ?PKC targets, and identified that the proteins whose phosphorylation was altered by selective activation of ?PKC were mostly mitochondrial proteins. Analysis of the mitochondrial phosphoproteome led to the identification of 55 spots, corresponding to 37 individual proteins, exclusively phosphorylated, in the presence of ??RACK. The majority of the proteins identified were involved in glucose and lipid metabolism, components of the respiratory chain as well as mitochondrial heat shock proteins.The protective effect of ?PKC during ischemia involves phosphorylation of several mitochondrial proteins involved in glucose and lipid metabolism and oxidative phosphorylation. Regulation of these metabolic pathways by ?PKC phosphorylation may lead to ?PKC-mediated cardioprotection induced by ??RACK.

    View details for DOI 10.1253/circj.CJ-11-1360

    View details for Web of Science ID 000305037600031

    View details for PubMedCentralID PMC3527096

  • Aerobic exercise training upregulates skeletal muscle calpain and ubiquitin-proteasome systems in healthy mice JOURNAL OF APPLIED PHYSIOLOGY Cunha, T. F., Moreira, J. B., Paixao, N. A., Campos, J. C., Monteiro, A. W., Bacurau, A. V., Bueno, C. R., Ferreira, J. C., Brum, P. C. 2012; 112 (11): 1839-1846

    Abstract

    Aerobic exercise training (AET) is an important mechanical stimulus that modulates skeletal muscle protein turnover, leading to structural rearrangement. Since the ubiquitin-proteasome system (UPS) and calpain system are major proteolytic pathways involved in protein turnover, we aimed to investigate the effects of intensity-controlled AET on the skeletal muscle UPS and calpain system and their association to training-induced structural adaptations. Long-lasting effects of AET were studied in C57BL/6J mice after 2 or 8 wk of AET. Plantaris cross-sectional area (CSA) and capillarization were assessed by myosin ATPase staining. mRNA and protein expression levels of main components of the UPS and calpain system were evaluated in plantaris by real-time PCR and Western immunoblotting, respectively. No proteolytic system activation was observed after 2 wk of AET. Eight weeks of AET resulted in improved running capacity, plantaris capillarization, and CSA. Muscle RING finger-1 mRNA expression was increased in 8-wk-trained mice. Accordingly, elevated 26S proteasome activity was observed in the 8-wk-trained group, without accumulation of ubiquitinated or carbonylated proteins. In addition, calpain abundance was increased by 8 wk of AET, whereas no difference was observed in its endogenous inhibitor calpastatin. Taken together, our findings indicate that skeletal muscle enhancements, as evidenced by increased running capacity, plantaris capillarization, and CSA, occurred in spite of the upregulated UPS and calpain system, suggesting that overactivation of skeletal muscle proteolytic systems is not restricted to atrophying states. Our data provide evidence for the contribution of the UPS and calpain system to metabolic turnover of myofibrillar proteins and skeletal muscle adaptations to AET.

    View details for DOI 10.1152/japplphysiol.00346.2011

    View details for Web of Science ID 000304810500006

    View details for PubMedID 22461440

  • Protein quality control disruption by PKCbetaII in heart failure Experimental Biology Meeting 2012 Ferreira, J. C., Mochly-Rosen, D., Brum, P. C. FEDERATION AMER SOC EXP BIOL. 2012
  • Protein Quality Control Disruption by PKC beta II in Heart Failure; Rescue by the Selective PKC beta II Inhibitor, beta IIV5-3 PLOS ONE Ferreira, J. C., Boer, B. N., Grinberg, M., Brum, P. C., Mochly-Rosen, D. 2012; 7 (3)

    Abstract

    Myocardial remodeling and heart failure (HF) are common sequelae of many forms of cardiovascular disease and a leading cause of mortality worldwide. Accumulation of damaged cardiac proteins in heart failure has been described. However, how protein quality control (PQC) is regulated and its contribution to HF development are not known. Here, we describe a novel role for activated protein kinase C isoform ?II (PKC?II) in disrupting PQC. We show that active PKC?II directly phosphorylated the proteasome and inhibited proteasomal activity in vitro and in cultured neonatal cardiomyocytes. Importantly, inhibition of PKC?II, using a selective PKC?II peptide inhibitor (?IIV5-3), improved proteasomal activity and conferred protection in cultured neonatal cardiomyocytes. We also show that sustained inhibition of PKC?II increased proteasomal activity, decreased accumulation of damaged and misfolded proteins and increased animal survival in two rat models of HF. Interestingly, ?IIV5-3-mediated protection was blunted by sustained proteasomal inhibition in HF. Finally, increased cardiac PKC?II activity and accumulation of misfolded proteins associated with decreased proteasomal function were found also in remodeled and failing human hearts, indicating a potential clinical relevance of our findings. Together, our data highlights PKC?II as a novel inhibitor of proteasomal function. PQC disruption by increased PKC?II activity in vivo appears to contribute to the pathophysiology of heart failure, suggesting that PKC?II inhibition may benefit patients with heart failure. (218 words).

    View details for DOI 10.1371/journal.pone.0033175

    View details for Web of Science ID 000305339100036

    View details for PubMedCentralID PMC3316563

  • Nitroglycerin use in myocardial infarction patients. Circulation journal Ferreira, J. C., Mochly-Rosen, D. 2012; 76 (1): 15-21

    Abstract

    Acute myocardial infarction (MI) and its sequelae are leading causes of morbidity and mortality worldwide. Nitroglycerin (glyceryl trinitrate [GTN]) remains a first-line treatment for angina pectoris and acute MI. Nitroglycerin achieves its benefit by giving rise to nitric oxide (NO), which causes vasodilation and increases blood flow to the myocardium. However, continuous delivery of GTN results in tolerance, limiting the use of this drug. Nitroglycerin tolerance is caused, at least in part, by inactivation of aldehyde dehydrogenase 2 (ALDH2), an enzyme that converts GTN to the vasodilator, NO. We recently found that in a MI model in animals, in addition to GTN's effect on the vasculature, sustained treatment negatively affected cardiomyocyte viability following ischemia, thus resulting in increased infarct size. Coadministration of Alda-1, an activator of ALDH2, with GTN improves metabolism of reactive aldehyde adducts and prevents the GTN-induced increase in cardiac dysfunction following MI. In this review, we describe the molecular mechanisms associated with the benefits and risks of GTN administration in MI.

    View details for PubMedID 22040938

  • Nitroglycerin Use in Myocardial Infarction Patients - Risks and Benefits CIRCULATION JOURNAL Ferreira, J. C., Mochly-Rosen, D. 2012; 76 (1): 15-21

    Abstract

    Acute myocardial infarction (MI) and its sequelae are leading causes of morbidity and mortality worldwide. Nitroglycerin (glyceryl trinitrate [GTN]) remains a first-line treatment for angina pectoris and acute MI. Nitroglycerin achieves its benefit by giving rise to nitric oxide (NO), which causes vasodilation and increases blood flow to the myocardium. However, continuous delivery of GTN results in tolerance, limiting the use of this drug. Nitroglycerin tolerance is caused, at least in part, by inactivation of aldehyde dehydrogenase 2 (ALDH2), an enzyme that converts GTN to the vasodilator, NO. We recently found that in a MI model in animals, in addition to GTN's effect on the vasculature, sustained treatment negatively affected cardiomyocyte viability following ischemia, thus resulting in increased infarct size. Coadministration of Alda-1, an activator of ALDH2, with GTN improves metabolism of reactive aldehyde adducts and prevents the GTN-induced increase in cardiac dysfunction following MI. In this review, we describe the molecular mechanisms associated with the benefits and risks of GTN administration in MI.

    View details for DOI 10.1253/circj.CJ-11-1133

    View details for Web of Science ID 000298771100004

    View details for PubMedCentralID PMC3527093

  • Regulation of cardiac excitability by protein kinase C isozymes. Frontiers in bioscience (Scholar edition) Ferreira, J. C., Mochly-Rosen, D., Boutjdir, M. 2012; 4: 532-546

    Abstract

    Cardiac excitability and electrical activity are determined by the sum of individual ion channels, gap junctions and exchanger activities. Electrophysiological remodeling during heart disease involves changes in membrane properties of cardiomyocytes and is related to higher prevalence of arrhythmia-associated morbidity and mortality. Pharmacological and genetic manipulation of cardiac cells as well as animal models of cardiovascular diseases are used to identity changes in electrophysiological properties and the molecular mechanisms associated with the disease. Protein kinase C (PKC) and several other kinases play a pivotal role in cardiac electrophysiological remodeling. Therefore, identifying specific therapies that regulate these kinases is the main focus of current research. PKC, a family of serine/threonine kinases, has been implicated as potential signaling nodes associated with biochemical and biophysical stress in cardiovascular diseases. In this review, we describe the role of PKC isozymes that are involved in cardiac excitability and discuss both genetic and pharmacological tools that were used, their attributes and limitations. Selective and effective pharmacological interventions to normalize cardiac electrical activities and correct cardiac arrhythmias will be of great clinical benefit.

    View details for PubMedID 22202075

  • Protein quality control disruption by PKCII in heart failure; rescue by the selective PKCII inhibitor, IIV5-3. PloS one Ferreira, J. C., Boer, B. N., Grinberg, M., Brum, P. C., Mochly-Rosen, D. 2012; 7 (3)

    Abstract

    Myocardial remodeling and heart failure (HF) are common sequelae of many forms of cardiovascular disease and a leading cause of mortality worldwide. Accumulation of damaged cardiac proteins in heart failure has been described. However, how protein quality control (PQC) is regulated and its contribution to HF development are not known. Here, we describe a novel role for activated protein kinase C isoform ?II (PKC?II) in disrupting PQC. We show that active PKC?II directly phosphorylated the proteasome and inhibited proteasomal activity in vitro and in cultured neonatal cardiomyocytes. Importantly, inhibition of PKC?II, using a selective PKC?II peptide inhibitor (?IIV5-3), improved proteasomal activity and conferred protection in cultured neonatal cardiomyocytes. We also show that sustained inhibition of PKC?II increased proteasomal activity, decreased accumulation of damaged and misfolded proteins and increased animal survival in two rat models of HF. Interestingly, ?IIV5-3-mediated protection was blunted by sustained proteasomal inhibition in HF. Finally, increased cardiac PKC?II activity and accumulation of misfolded proteins associated with decreased proteasomal function were found also in remodeled and failing human hearts, indicating a potential clinical relevance of our findings. Together, our data highlights PKC?II as a novel inhibitor of proteasomal function. PQC disruption by increased PKC?II activity in vivo appears to contribute to the pathophysiology of heart failure, suggesting that PKC?II inhibition may benefit patients with heart failure. (218 words).

    View details for DOI 10.1371/journal.pone.0033175

    View details for PubMedID 22479367

  • Identification of ePKC targets during cardiac ischemic injury. Circulation journal Budas, G., Costa, H. M., Ferreira, J. C., Teixeira da Silva Ferreira, A., Perales, J., Krieger, J. E., Mochly-Rosen, D., Schechtman, D. 2012; 76 (6): 1476-1485

    Abstract

    Epsilon-protein kinase C (?PKC) protects the heart from ischemic injury. However, the mechanism(s) of ?PKC cardioprotection is still unclear. Identification of the ?PKC targets may aid in elucidating the ?PKC-mediated cardioprotective mechanisms. Previous studies, using ?PKC transgenic mice and difference in gel electrophoresis, identified proteins involved in glucose metabolism, the expression of which was modified by ?PKC. Those studies were accompanied by metabolomic analysis, suggesting that increased glucose oxidation may be responsible for the cardioprotective effect of ?PKC. Whether these ?PKC-mediated alterations were because of differences in protein expression or phosphorylation was not determined.In the present study, we used an ?PKC -specific activator peptide, ??RACK, combined with phosphoproteomics, to find ?PKC targets, and identified that the proteins whose phosphorylation was altered by selective activation of ?PKC were mostly mitochondrial proteins. Analysis of the mitochondrial phosphoproteome led to the identification of 55 spots, corresponding to 37 individual proteins, exclusively phosphorylated, in the presence of ??RACK. The majority of the proteins identified were involved in glucose and lipid metabolism, components of the respiratory chain as well as mitochondrial heat shock proteins.The protective effect of ?PKC during ischemia involves phosphorylation of several mitochondrial proteins involved in glucose and lipid metabolism and oxidative phosphorylation. Regulation of these metabolic pathways by ?PKC phosphorylation may lead to ?PKC-mediated cardioprotection induced by ??RACK.

    View details for PubMedID 22453000

  • Pharmacological inhibition of beta IIPKC is cardioprotective in late-stage hypertrophy JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY Ferreira, J. C., Koyanagi, T., Palaniyandi, S. s., Fajardo, G., Churchill, E. N., Budas, G., Disatnik, M., Bernstein, D., Brum, P. C., Mochly-Rosen, D. 2011; 51 (6): 980-987

    Abstract

    We previously found that in the hearts of hypertensive Dahl salt-sensitive rats, ?IIPKC levels increase during the transition from compensated cardiac hypertrophy to cardiac dysfunction. Here we showed that a six-week treatment of these hypertensive rats with a ?IIPKC-specific inhibitor, ?IIV5-3, prolonged their survival by at least 6weeks, suppressed myocardial fibrosis and inflammation, and delayed the transition from compensated hypertrophy to cardiac dysfunction. In addition, changes in the levels of the Ca(2+)-handling proteins, SERCA2 and the Na(+)/Ca(2+) exchanger, as well as troponin I phosphorylation, seen in the control-treated hypertensive rats were not observed in the ???PKC-treated rats, suggesting that ???PKC contributes to the regulation of calcium levels in the myocardium. In contrast, treatment with the selective inhibitor of ?IPKC, an alternative spliced form of ?IIPKC, had no beneficial effects in these rats. We also found that ?IIV5-3, but not ?IV5-3, improved calcium handling in isolated rat cardiomyocytes and enhanced contractility in isolated rat hearts. In conclusion, our data using an in vivo model of cardiac dysfunction (late-phase hypertrophy), suggest that ?IIPKC contributes to the pathology associated with heart failure and thus an inhibitor of ?IIPKC may be a potential treatment for this disease.

    View details for DOI 10.1016/j.yjmcc.2011.08.025

    View details for PubMedID 21920368

  • ALDH2 activator inhibits increased myocardial infarction injury by nitroglycerin tolerance. Science translational medicine Sun, L., Ferreira, J. C., Mochly-Rosen, D. 2011; 3 (107): 107ra111-?

    Abstract

    Nitroglycerin, which treats impaired cardiac function through vasodilation as it is converted to nitric oxide, is used worldwide for patients with various ischemic and congestive cardiac diseases, including angina pectoris. Nevertheless, after continuous treatment, the benefits of nitroglycerin are limited by the development of tolerance to the drug. Nitroglycerin tolerance is a result of inactivation of aldehyde dehydrogenase 2 (ALDH2), an enzyme essential for cardioprotection in animals subjected to myocardial infarction. Here, we tested the hypothesis that the tolerance that develops as a result of sustained nitroglycerin treatment increases cardiac injury by subsequent myocardial infarction. In a rat model of myocardial infarction, 16 hours of prior, sustained nitroglycerin treatment resulted in infarcts that were twice as large as those in untreated control animals and in diminished cardiac function at 3 days and 2 weeks after the myocardial infarction. We also sought to identify a potential treatment to protect against this increased cardiac damage. Nitroglycerin inhibited ALDH2 activity in vitro, an effect that was blocked by Alda-1, an activator of ALDH2. Co-administration of Alda-1 with the nitroglycerin prevented the nitroglycerin-induced increase in cardiac dysfunction after myocardial infarction in rats, at least in part by enhancing metabolism of reactive aldehyde adducts that impair normal protein functions. If our animal studies showing that nitroglycerin tolerance increases cardiac injury upon ischemic insult are corroborated in humans, activators of ALDH2 such as Alda-1 may help to protect patients with myocardial infarction from this nitroglycerin-induced increase in cardiac injury while maintaining the cardiac benefits of the increased nitric oxide concentrations produced by nitroglycerin.

    View details for DOI 10.1126/scitranslmed.3002067

    View details for PubMedID 22049071

  • ALDH2 Activator Inhibits Increased Myocardial Infarction Injury by Nitroglycerin Tolerance SCIENCE TRANSLATIONAL MEDICINE Sun, L., Cesar, J., Ferreira, B., Mochly-Rosen, D. 2011; 3 (107)

    Abstract

    Nitroglycerin, which treats impaired cardiac function through vasodilation as it is converted to nitric oxide, is used worldwide for patients with various ischemic and congestive cardiac diseases, including angina pectoris. Nevertheless, after continuous treatment, the benefits of nitroglycerin are limited by the development of tolerance to the drug. Nitroglycerin tolerance is a result of inactivation of aldehyde dehydrogenase 2 (ALDH2), an enzyme essential for cardioprotection in animals subjected to myocardial infarction. Here, we tested the hypothesis that the tolerance that develops as a result of sustained nitroglycerin treatment increases cardiac injury by subsequent myocardial infarction. In a rat model of myocardial infarction, 16 hours of prior, sustained nitroglycerin treatment resulted in infarcts that were twice as large as those in untreated control animals and in diminished cardiac function at 3 days and 2 weeks after the myocardial infarction. We also sought to identify a potential treatment to protect against this increased cardiac damage. Nitroglycerin inhibited ALDH2 activity in vitro, an effect that was blocked by Alda-1, an activator of ALDH2. Co-administration of Alda-1 with the nitroglycerin prevented the nitroglycerin-induced increase in cardiac dysfunction after myocardial infarction in rats, at least in part by enhancing metabolism of reactive aldehyde adducts that impair normal protein functions. If our animal studies showing that nitroglycerin tolerance increases cardiac injury upon ischemic insult are corroborated in humans, activators of ALDH2 such as Alda-1 may help to protect patients with myocardial infarction from this nitroglycerin-induced increase in cardiac injury while maintaining the cardiac benefits of the increased nitric oxide concentrations produced by nitroglycerin.

    View details for DOI 10.1126/scitranslmed.3002067

    View details for Web of Science ID 000296750400005

    View details for PubMedCentralID PMC3547591

  • IIPKC and ePKC isozymes as potential pharmacological targets in cardiac hypertrophy and heart failure. Journal of molecular and cellular cardiology Ferreira, J. C., Brum, P. C., Mochly-Rosen, D. 2011; 51 (4): 479-484

    Abstract

    Cardiac hypertrophy is a complex adaptive response to mechanical and neurohumoral stimuli and under continual stressor, it contributes to maladaptive responses, heart failure and death. Protein kinase C (PKC) and several other kinases play a role in the maladaptative cardiac responses, including cardiomyocyte hypertrophy, myocardial fibrosis and inflammation. Identifying specific therapies that regulate these kinases is a major focus of current research. PKC, a family of serine/threonine kinases, has emerged as potential mediators of hypertrophic stimuli associated with neurohumoral hyperactivity in heart failure. In this review, we describe the role of PKC isozymes that is involved in cardiac hypertrophy and heart failure. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure".

    View details for DOI 10.1016/j.yjmcc.2010.10.020

    View details for PubMedID 21035454

  • beta IIPKC and epsilon PKC isozymes as potential pharmacological targets in cardiac hypertrophy and heart failure JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY Batista Ferreira, J. C., Brum, P. C., Mochly-Rosen, D. 2011; 51 (4): 479-484

    Abstract

    Cardiac hypertrophy is a complex adaptive response to mechanical and neurohumoral stimuli and under continual stressor, it contributes to maladaptive responses, heart failure and death. Protein kinase C (PKC) and several other kinases play a role in the maladaptative cardiac responses, including cardiomyocyte hypertrophy, myocardial fibrosis and inflammation. Identifying specific therapies that regulate these kinases is a major focus of current research. PKC, a family of serine/threonine kinases, has emerged as potential mediators of hypertrophic stimuli associated with neurohumoral hyperactivity in heart failure. In this review, we describe the role of PKC isozymes that is involved in cardiac hypertrophy and heart failure. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure".

    View details for DOI 10.1016/j.yjmcc.2010.10.020

    View details for Web of Science ID 000295302900011

    View details for PubMedCentralID PMC3135714

  • Aerobic exercise training in heart failure: impact on sympathetic hyperactivity and cardiac and skeletal muscle function Brum, P. C., Bacurau, A. V., Medeiros, A., Ferreira, J. C., Vanzelli, A. S., Negrao, C. E. ASSOC BRAS DIVULG CIENTIFICA. 2011: 827-835

    Abstract

    Heart failure is a common endpoint for many forms of cardiovascular disease and a significant cause of morbidity and mortality. Chronic neurohumoral excitation (i.e., sympathetic hyperactivity) has been considered to be a hallmark of heart failure and is associated with a poor prognosis, cardiac dysfunction and remodeling, and skeletal myopathy. Aerobic exercise training is efficient in counteracting sympathetic hyperactivity and its toxic effects on cardiac and skeletal muscles. In this review, we describe the effects of aerobic exercise training on sympathetic hyperactivity, skeletal myopathy, as well as cardiac function and remodeling in human and animal heart failure. We also discuss the mechanisms underlying the effects of aerobic exercise training.

    View details for Web of Science ID 000295721500002

    View details for PubMedID 21956529

  • PKCII inhibition attenuates myocardial infarction induced heart failure and is associated with a reduction of fibrosis and pro-inflammatory responses. Journal of cellular and molecular medicine Palaniyandi, S. S., Ferreira, J. C., Brum, P. C., Mochly-Rosen, D. 2011; 15 (8): 1769-1777

    Abstract

    Protein kinase C ?II (PKC?II) levels increase in the myocardium of patients with end-stage heart failure (HF). Also targeted overexpression of PKC?II in the myocardium of mice leads to dilated cardiomyopathy associated with inflammation, fibrosis and myocardial dysfunction. These reports suggest a deleterious role of PKC?II in HF development. Using a post-myocardial infarction (MI) model of HF in rats, we determined the benefit of chronic inhibition of PKC?II on the progression of HF over a period of 6 weeks after the onset of symptoms and the cellular basis for these effects. Four weeks after MI, rats with HF signs that were treated for 6 weeks with the PKC?II selective inhibitor (?IIV5-3 conjugated to TAT(47-57) carrier peptide) (3 mg/kg/day) showed improved fractional shortening (from 21% to 35%) compared to control (TAT(47-57) carrier peptide alone). Formalin-fixed mid-ventricle tissue sections stained with picrosirius red, haematoxylin and eosin and toluidine blue dyes exhibited a 150% decrease in collagen deposition, a two-fold decrease in inflammation and a 30% reduction in mast cell degranulation, respectively, in rat hearts treated with the selective PKC?II inhibitor. Further, a 90% decrease in active TGF?1 and a significant reduction in SMAD2/3 phosphorylation indicated that the selective inhibition of PKC?II attenuates cardiac remodelling mediated by the TGF-SMAD signalling pathway. Therefore, sustained selective inhibition of PKC?II in a post-MI HF rat model improves cardiac function and is associated with inhibition of pathological myocardial remodelling.

    View details for DOI 10.1111/j.1582-4934.2010.01174.x

    View details for PubMedID 20874717

  • PKC beta II inhibition attenuates myocardial infarction induced heart failure and is associated with a reduction of fibrosis and pro-inflammatory responses JOURNAL OF CELLULAR AND MOLECULAR MEDICINE Palaniyandi, S. S., Batista Ferreira, J. C., Brum, P. C., Mochly-Rosen, D. 2011; 15 (8): 1769-1777

    Abstract

    Protein kinase C ?II (PKC?II) levels increase in the myocardium of patients with end-stage heart failure (HF). Also targeted overexpression of PKC?II in the myocardium of mice leads to dilated cardiomyopathy associated with inflammation, fibrosis and myocardial dysfunction. These reports suggest a deleterious role of PKC?II in HF development. Using a post-myocardial infarction (MI) model of HF in rats, we determined the benefit of chronic inhibition of PKC?II on the progression of HF over a period of 6 weeks after the onset of symptoms and the cellular basis for these effects. Four weeks after MI, rats with HF signs that were treated for 6 weeks with the PKC?II selective inhibitor (?IIV5-3 conjugated to TAT(47-57) carrier peptide) (3 mg/kg/day) showed improved fractional shortening (from 21% to 35%) compared to control (TAT(47-57) carrier peptide alone). Formalin-fixed mid-ventricle tissue sections stained with picrosirius red, haematoxylin and eosin and toluidine blue dyes exhibited a 150% decrease in collagen deposition, a two-fold decrease in inflammation and a 30% reduction in mast cell degranulation, respectively, in rat hearts treated with the selective PKC?II inhibitor. Further, a 90% decrease in active TGF?1 and a significant reduction in SMAD2/3 phosphorylation indicated that the selective inhibition of PKC?II attenuates cardiac remodelling mediated by the TGF-SMAD signalling pathway. Therefore, sustained selective inhibition of PKC?II in a post-MI HF rat model improves cardiac function and is associated with inhibition of pathological myocardial remodelling.

    View details for DOI 10.1111/j.1582-4934.2010.01174.x

    View details for Web of Science ID 000292700800012

    View details for PubMedCentralID PMC3136735

  • Creatine in Type 2 Diabetes: A Randomized, Double-Blind, Placebo-Controlled Trial MEDICINE AND SCIENCE IN SPORTS AND EXERCISE Gualano, B., Painneli, V. D., Roschel, H., Artioli, G. G., Neves, M., de Sa Pinto, A. L., Rossi da Silva, M. E., Cunha, M. R., Garcia Otaduy, M. C., Leite, C. d., Ferreira, J. C., Pereira, R. M., Brum, P. C., Bonfa, E., Lancha, A. H. 2011; 43 (5): 770-778

    Abstract

    Creatine supplementation improves glucose tolerance in healthy subjects.The aim was to investigate whether creatine supplementation has a beneficial effect on glycemic control of type 2 diabetic patients undergoing exercise training.A 12-wk randomized, double-blind, placebo-controlled trial was performed. The patients were allocated to receive either creatine (CR) (5 gd) or placebo (PL) and were enrolled in an exercise training program. The primary outcome was glycosylated hemoglobin (HbA1c). Secondary outcomes included the area under the curve of glucose, insulin, and C-peptide and insulin sensitivity indexes. Physical capacity, lipid profile, and GLUT-4 protein expression and translocation were also assessed.Twenty-five subjects were analyzed (CR: n=13; PL: n=12). HbA1c was significantly reduced in the creatine group when compared with the placebo group (CR: PRE=7.4 0.7, POST=6.4 0.4; PL: PRE=7.5 0.6, POST=7.6 0.7; P=0.004; difference=-1.1%, 95% confidence interval=-1.9% to -0.4%). The delta area under the curve of glucose concentration was significantly lower in the CR group than in the PL group (CR=-7790 4600, PL=2008 7614; P=0.05). The CR group also presented decreased glycemia at times 0, 30, and 60 min during a meal tolerance test and increased GLUT-4 translocation. Insulin and C-peptide concentrations, surrogates of insulin sensitivity, physical capacity, lipid profile, and adverse effects were comparable between the groups.Creatine supplementation combined with an exercise program improves glycemic control in type 2 diabetic patients. The underlying mechanism seems to be related to an increase in GLUT-4 recruitment to the sarcolemma.

    View details for DOI 10.1249/MSS.0b013e3181fcee7d

    View details for Web of Science ID 000289557100005

    View details for PubMedID 20881878

  • Angiotensin receptor blockade improves the net balance of cardiac Ca2+ handling-related proteins in sympathetic hyperactivity-induced heart failure LIFE SCIENCES Ferreira, J. C., Moreira, J. B., Campos, J. C., Pereira, M. G., Mattos, K. C., Coelho, M. A., Brum, P. C. 2011; 88 (13-14): 578-585

    Abstract

    The clinical benefits of angiotensin II type 1 (AT1) receptor blockers (ARB) in heart failure (HF) include cardiac anti-remodeling and improved ventricular function. However, the cellular mechanisms underlying the benefits of ARB on ventricular function need to be better clarified. In the present manuscript, we evaluated the effects of AT1 receptor blockade on the net balance of Ca(2+) handling proteins in hearts of mice lacking ?(2A) and ?(2C) adrenoceptors (?(2A)/?(2C)ARKO), which develop sympathetic hyperactivity (SH) induced-HF.A cohort of male wild-type (WT) and congenic ?(2A)/?(2C)ARKO mice in a C57BL6/J genetic background (5-7mo of age) was randomly assigned to receive either placebo or ARB (Losartan, 10mg/kg for 8wks). Ventricular function (VF) was assessed by echocardiography, and cardiac myocyte width and ventricular fibrosis by a computer-assisted morphometric system. Sarcoplasmic reticulum Ca(2+) ATPase (SERCA2), phospholamban (PLN), phospho-Ser(16)-PLN, phospho-Thr(17)-PLN, phosphatase 1 (PP1), Na(+)-Ca(2+) exchanger (NCX), Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and phospho-Thr(286)-CaMKII were analyzed by Western blot.?(2A)/?(2C)ARKO mice displayed ventricular dysfunction, cardiomyocyte hypertrophy and cardiac fibrosis paralleled by decreased SERCA2 and increased phospho-Thr(17)-PLN, CaMKII, phospho-Thr(286)-CaMKII and NCX levels. ARB induced anti-cardiac remodeling effect and improved VF in ?(2A)/?(2C)ARKO associated with increased SERCA2 and phospho-Ser(16)-PLN levels, and SERCA2:NCX ratio. Additionally, ARB decreased phospho-Thr(17)-PLN levels as well as reestablished NCX, CaMKII and phospho-Thr(286)-CaMKII toward WT levels.Altogether, these data provide new insights on intracellular Ca(2+) regulatory mechanisms underlying improved ventricular function by ARB therapy in HF.

    View details for DOI 10.1016/j.lfs.2011.01.009

    View details for Web of Science ID 000288819500003

    View details for PubMedID 21277865

  • Exercise Training and Caloric Restriction Prevent Reduction in Cardiac Ca2+-Handling Protein Profile in Obese Rats HYPERTENSION Paulino, E. C., Batista Ferreira, J. C., Bechara, L. R., Tsutsui, J. M., Mathias, W., Lima, F. B., Casarini, D. E., Cicogna, A. C., Brum, P. C., Negrao, C. E. 2010; 56 (4): 629-U137

    Abstract

    Previous studies show that exercise training and caloric restriction improve cardiac function in obesity. However, the molecular mechanisms underlying this effect on cardiac function remain unknown. Thus, we studied the effect of exercise training and/or caloric restriction on cardiac function and Ca(2+) handling protein expression in obese rats. To accomplish this goal, male rats fed with a high-fat and sucrose diet for 25 weeks were randomly assigned into 4 groups: high-fat and sucrose diet, high-fat and sucrose diet and exercise training, caloric restriction, and exercise training and caloric restriction. An additional lean group was studied. The study was conducted for 10 weeks. Cardiac function was evaluated by echocardiography and Ca(2+) handling protein expression by Western blotting. Our results showed that visceral fat mass, circulating leptin, epinephrine, and norepinephrine levels were higher in rats on the high-fat and sucrose diet compared with the lean rats. Cardiac nitrate levels, reduced/oxidized glutathione, left ventricular fractional shortening, and protein expression of phosphorylated Ser(2808)-ryanodine receptor and Thr(17)-phospholamban were lower in rats on the high-fat and sucrose diet compared with lean rats. Exercise training and/or caloric restriction prevented increases in visceral fat mass, circulating leptin, epinephrine, and norepinephrine levels and prevented reduction in cardiac nitrate levels and reduced:oxidized glutathione ratio. Exercise training and/or caloric restriction prevented reduction in left ventricular fractional shortening and in phosphorylation of the Ser(2808)-ryanodine receptor and Thr(17)-phospholamban. These findings show that exercise training and/or caloric restriction prevent cardiac dysfunction in high-fat and sucrose diet rats, which seems to be attributed to decreased circulating neurohormone levels. In addition, this nonpharmacological paradigm prevents a reduction in the Ser(2808)-ryanodine receptor and Thr(17)-phospholamban phosphorylation and redox status.

    View details for DOI 10.1161/HYPERTENSIONAHA.110.156141

    View details for Web of Science ID 000281881400015

    View details for PubMedID 20644006

  • Aerobic exercise training improves skeletal muscle function and Ca2+ handling-related protein expression in sympathetic hyperactivity-induced heart failure JOURNAL OF APPLIED PHYSIOLOGY Bueno, C. R., Ferreira, J. C., Pereira, M. G., Bacurau, A. V., Brum, P. C. 2010; 109 (3): 702-709

    Abstract

    The cellular mechanisms of positive effects associated with aerobic exercise training on overall intrinsic skeletal muscle changes in heart failure (HF) remain unclear. We investigated potential Ca2+ abnormalities in skeletal muscles comprising different fiber compositions and investigated whether aerobic exercise training would improve muscle function in a genetic model of sympathetic hyperactivity-induced HF. A cohort of male 5-mo-old wild-type (WT) and congenic alpha2A/alpha2C adrenoceptor knockout (ARKO) mice in a C57BL/6J genetic background were randomly assigned into untrained and trained groups. Exercise training consisted of a 8-wk running session of 60 min, 5 days/wk (from 5 to 7 mo of age). After completion of the exercise training protocol, exercise tolerance was determined by graded treadmill exercise test, muscle function test by Rotarod, ambulation and resistance to inclination tests, cardiac function by echocardiography, and Ca2+ handling-related protein expression by Western blot. alpha2A/alpha2CARKO mice displayed decreased ventricular function, exercise intolerance, and muscle weakness paralleled by decreased expression of sarcoplasmic Ca2+ release-related proteins [alpha1-, alpha2-, and beta1-subunits of dihydropyridine receptor (DHPR) and ryanodine receptor (RyR)] and Ca2+ reuptake-related proteins [sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)1/2 and Na+/Ca2+ exchanger (NCX)] in soleus and plantaris. Aerobic exercise training significantly improved exercise tolerance and muscle function and reestablished the expression of proteins involved in sarcoplasmic Ca2+ handling toward WT levels. We provide evidence that Ca2+ handling-related protein expression is decreased in this HF model and that exercise training improves skeletal muscle function associated with changes in the net balance of skeletal muscle Ca2+ handling proteins.

    View details for DOI 10.1152/japplphysiol.00281.2010

    View details for Web of Science ID 000281958800012

    View details for PubMedID 20595538

  • Regulation of mitochondrial processes: a target for heart failure. Drug discovery today. Disease mechanisms Palaniyandi, S. S., Qi, X., Yogalingam, G., Ferreira, J. C., Mochly-Rosen, D. 2010; 7 (2): e95-e102

    Abstract

    Cardiac mitochondria, the main source of energy as well as free radicals, are vital organelles for normal functioning of the heart. Mitochondrial number, structure, turnover and function are regulated by processes such as mitochondrial protein quality control, mitochondrial fusion and fission and mitophagy. Recent studies suggest that abnormal changes in these mitochondrial regulatory processes may contribute to the pathology of heart failure (HF). Here we discuss these processes and their potential as therapeutic targets.

    View details for PubMedID 21278905

  • Aerobic exercise training improves Ca2+ handling and redox status of skeletal muscle in mice EXPERIMENTAL BIOLOGY AND MEDICINE Ferreira, J. C., Bacurau, A. V., Bueno Junior, C. R., Cunha, T. C., Tanaka, L. Y., Jardim, M. A., Ramires, P. R., Brum, P. C. 2010; 235 (4): 497-505

    Abstract

    Exercise training is known to promote relevant changes in the properties of skeletal muscle contractility toward powerful fibers. However, there are few studies showing the effect of a well-established exercise training protocol on Ca(2+) handling and redox status in skeletal muscles with different fiber-type compositions. We have previously standardized a valid and reliable protocol to improve endurance exercise capacity in mice based on maximal lactate steady-state workload (MLSSw). The aim of this study was to investigate the effect of exercise training, performed at MLSSw, on the skeletal muscle Ca(2+) handling-related protein levels and cellular redox status in soleus and plantaris. Male C57BL/6J mice performed treadmill training at MLSSw over a period of eight weeks. Muscle fiber-typing was determined by myosin ATPase histochemistry, citrate synthase activity by spectrophotometric assay, Ca(2+) handling-related protein levels by Western blot and reduced to oxidized glutathione ratio (GSH:GSSG) by high-performance liquid chromatography. Trained mice displayed higher running performance and citrate synthase activity compared with untrained mice. Improved running performance in trained mice was paralleled by fast-to-slow fiber-type shift and increased capillary density in both plantaris and soleus. Exercise training increased dihydropyridine receptor (DHPR) alpha2 subunit, ryanodine receptor and Na(+)/Ca(2+) exchanger levels in plantaris and soleus. Moreover, exercise training elevated DHPR beta1 subunit and sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) 1 levels in plantaris and SERCA2 levels in soleus of trained mice. Skeletal muscle GSH content and GSH:GSSG ratio was increased in plantaris and soleus of trained mice. Taken together, our findings indicate that MLSSw exercise-induced better running performance is, in part, due to increased levels of proteins involved in skeletal muscle Ca(2+) handling, whereas this response is partially dependent on specificity of skeletal muscle fiber-type composition. Finally, we demonstrated an augmented cellular redox status and GSH antioxidant capacity in trained mice.

    View details for DOI 10.1258/ebm.2009.009165

    View details for Web of Science ID 000277691700011

    View details for PubMedID 20407082

  • Ischaemic preconditioning improves proteasomal activity and increases the degradation of delta PKC during reperfusion CARDIOVASCULAR RESEARCH Churchill, E. N., Ferreira, J. C., Brum, P. C., Szweda, L. I., Mochly-Rosen, D. 2010; 85 (2): 385-394

    Abstract

    The response of the myocardium to an ischaemic insult is regulated by two highly homologous protein kinase C (PKC) isozymes, delta and epsilonPKC. Here, we determined the spatial and temporal relationships between these two isozymes in the context of ischaemia/reperfusion (I/R) and ischaemic preconditioning (IPC) to better understand their roles in cardioprotection.Using an ex vivo rat model of myocardial infarction, we found that short bouts of ischaemia and reperfusion prior to the prolonged ischaemic event (IPC) diminished deltaPKC translocation by 3.8-fold and increased epsilonPKC accumulation at mitochondria by 16-fold during reperfusion. In addition, total cellular levels of deltaPKC decreased by 60 +/- 2.7% in response to IPC, whereas the levels of epsilonPKC did not significantly change. Prolonged ischaemia induced a 48 +/- 11% decline in the ATP-dependent proteasomal activity and increased the accumulation of misfolded proteins during reperfusion by 192 +/- 32%; both of these events were completely prevented by IPC. Pharmacological inhibition of the proteasome or selective inhibition of epsilonPKC during IPC restored deltaPKC levels at the mitochondria while decreasing epsilonPKC levels, resulting in a loss of IPC-induced protection from I/R. Importantly, increased myocardial injury was the result, in part, of restoring a deltaPKC-mediated I/R pro-apoptotic phenotype by decreasing pro-survival signalling and increasing cytochrome c release into the cytosol.Taken together, our findings indicate that IPC prevents I/R injury at reperfusion by protecting ATP-dependent 26S proteasomal function. This decreases the accumulation of the pro-apoptotic kinase, deltaPKC, at cardiac mitochondria, resulting in the accumulation of the pro-survival kinase, epsilonPKC.

    View details for DOI 10.1093/cvr/cvp334

    View details for PubMedID 19820255

  • Cardiac anti-remodelling effect of aerobic training is associated with a reduction in the calcineurin/NFAT signalling pathway in heart failure mice JOURNAL OF PHYSIOLOGY-LONDON Oliveira, R. S., Ferreira, J. C., Gomes, E. R., Paixao, N. A., Rolim, N. P., Medeiros, A., Guatimosim, S., Brum, P. C. 2009; 587 (15): 3899-3910

    Abstract

    Cardiomyocyte hypertrophy occurs in response to a variety of physiological and pathological stimuli. While pathological hypertrophy in heart failure is usually coupled with depressed contractile function, physiological hypertrophy associates with increased contractility. In the present study, we explored whether 8 weeks of moderate intensity exercise training would lead to a cardiac anti-remodelling effect in an experimental model of heart failure associated with a deactivation of a pathological (calcineurin/NFAT, CaMKII/HDAC) or activation of a physiological (Akt-mTOR) hypertrophy signalling pathway. The cardiac dysfunction, exercise intolerance, left ventricle dilatation, increased heart weight and cardiomyocyte hypertrophy from mice lacking alpha(2A) and alpha(2C) adrenoceptors (alpha(2A)/alpha(2C)ARKO mice) were associated with sympathetic hyperactivity induced heart failure. The relative contribution of Ca(2+)-calmodulin high-affinity (calcineurin/NFAT) and low-affinity (CaMKII/HDAC) targets to pathological hypertrophy of alpha(2A)/alpha(2C)ARKO mice was verified. While nuclear calcineurin B, NFATc3 and GATA-4 translocation were significantly increased in alpha(2A)/alpha(2C)ARKO mice, no changes were observed in CaMKII/HDAC activation. As expected, cyclosporine treatment decreased nuclear translocation of calcineurin/NFAT in alpha(2A)/alpha(2C)ARKO mice, which was associated with improved ventricular function and a pronounced anti-remodelling effect. The Akt/mTOR signalling pathway was not activated in alpha(2A)/alpha(2C)ARKO mice. Exercise training improved cardiac function and exercise capacity in alpha(2A)/alpha(2C)ARKO mice and decreased heart weight and cardiomyocyte width paralleled by diminished nuclear NFATc3 and GATA-4 translocation as well as GATA-4 expression levels. When combined, these findings support the notion that deactivation of calcineurin/NFAT pathway-induced pathological hypertrophy is a preferential mechanism by which exercise training leads to the cardiac anti-remodelling effect in heart failure.

    View details for DOI 10.1113/jphysiol.2009.173948

    View details for Web of Science ID 000268590300013

    View details for PubMedID 19505981

  • Protein kinase C in heart failure: a therapeutic target? CARDIOVASCULAR RESEARCH Palaniyandi, S. S., Sun, L., Batista Ferreira, J. C., Mochly-Rosen, D. 2009; 82 (2): 229-239

    Abstract

    Heart failure (HF) afflicts about 5 million people and causes 300,000 deaths a year in the United States alone. An integral part of the pathogenesis of HF is cardiac remodelling, and the signalling events that regulate it are a subject of intense research. Cardiac remodelling is the sum of responses of the heart to causes of HF, such as ischaemia, myocardial infarction, volume and pressure overload, infection, inflammation, and mechanical injury. These responses, including cardiomyocyte hypertrophy, myocardial fibrosis, and inflammation, involve numerous cellular and structural changes and ultimately result in a progressive decline in cardiac performance. Pharmacological and genetic manipulation of cultured heart cells and animal models of HF and the analysis of cardiac samples from patients with HF are all used to identify the molecular and cellular mechanisms leading to the disease. Protein kinase C (PKC) isozymes, a family of serine-threonine protein kinase enzymes, were found to regulate a number of cardiac responses, including those associated with HF. In this review, we describe the PKC isozymes that play critical roles in specific aspects of cardiac remodelling and dysfunction in HF.

    View details for DOI 10.1093/cvr/cvp001

    View details for PubMedID 19168855

  • Sympathetic hyperactivity differentially affects skeletal muscle mass in developing heart failure: role of exercise training JOURNAL OF APPLIED PHYSIOLOGY Bacurau, A. V., Jardim, M. A., Ferreira, J. C., Bechara, L. R., Bueno, C. R., Alba-Loureiro, T. C., Negrao, C. E., Casarini, D. E., Curi, R., Ramires, P. R., Moriscot, A. S., Brum, P. C. 2009; 106 (5): 1631-1640

    Abstract

    Sympathetic hyperactivity (SH) is a hallmark of heart failure (HF), and several lines of evidence suggest that SH contributes to HF-induced skeletal myopathy. However, little is known about the influence of SH on skeletal muscle morphology and metabolism in a setting of developing HF, taking into consideration muscles with different fiber compositions. The contribution of SH on exercise tolerance and skeletal muscle morphology and biochemistry was investigated in 3- and 7-mo-old mice lacking both alpha(2A)- and alpha(2C)-adrenergic receptor subtypes (alpha(2A)/alpha(2C)ARKO mice) that present SH with evidence of HF by 7 mo. To verify whether exercise training (ET) would prevent skeletal muscle myopathy in advanced-stage HF, alpha(2A)/alpha(2C)ARKO mice were exercised from 5 to 7 mo of age. At 3 mo, alpha(2A)/alpha(2C)ARKO mice showed no signs of HF and preserved exercise tolerance and muscular norepinephrine with no changes in soleus morphology. In contrast, plantaris muscle of alpha(2A)/alpha(2C)ARKO mice displayed hypertrophy and fiber type shift (IIA --> IIX) paralleled by capillary rarefaction, increased hexokinase activity, and oxidative stress. At 7 mo, alpha(2A)/alpha(2C)ARKO mice displayed exercise intolerance and increased muscular norepinephrine, muscular atrophy, capillary rarefaction, and increased oxidative stress. ET reestablished alpha(2A)/alpha(2C)ARKO mouse exercise tolerance to 7-mo-old wild-type levels and prevented muscular atrophy and capillary rarefaction associated with reduced oxidative stress. Collectively, these data provide direct evidence that SH is a major factor contributing to skeletal muscle morphological changes in a setting of developing HF. ET prevented skeletal muscle myopathy in alpha(2A)/alpha(2C)ARKO mice, which highlights its importance as a therapeutic tool for HF.

    View details for DOI 10.1152/japplphysiol.91067.2008

    View details for Web of Science ID 000265745500020

    View details for PubMedID 19179649

  • Exercise training reduces cardiac angiotensin II levels and prevents cardiac dysfunction in a genetic model of sympathetic hyperactivity-induced heart failure in mice EUROPEAN JOURNAL OF APPLIED PHYSIOLOGY Pereira, M. G., Ferreira, J. C., Bueno, C. R., Mattos, K. C., Rosa, K. T., Irigoyen, M. C., Oliveira, E. M., Krieger, J. E., Brum, P. C. 2009; 105 (6): 843-850

    Abstract

    The role of exercise training (ET) on cardiac renin-angiotensin system (RAS) was investigated in 3-5 month-old mice lacking alpha(2A-) and alpha(2C-)adrenoceptors (alpha(2A)/alpha(2C)ARKO) that present heart failure (HF) and wild type control (WT). ET consisted of 8-week running sessions of 60 min, 5 days/week. In addition, exercise tolerance, cardiac structural and function analysis were made. At 3 months, fractional shortening and exercise tolerance were similar between groups. At 5 months, alpha(2A)/alpha(2C)ARKO mice displayed ventricular dysfunction and fibrosis associated with increased cardiac angiotensin (Ang) II levels (2.9-fold) and increased local angiotensin-converting enzyme activity (ACE 18%). ET decreased alpha(2A)/alpha(2C)ARKO cardiac Ang II levels and ACE activity to age-matched untrained WT mice levels while increased ACE2 expression and prevented exercise intolerance and ventricular dysfunction with little impact on cardiac remodeling. Altogether, these data provide evidence that reduced cardiac RAS explains, at least in part, the beneficial effects of ET on cardiac function in a genetic model of HF.

    View details for DOI 10.1007/s00421-008-0967-4

    View details for Web of Science ID 000264175900003

    View details for PubMedID 19125280

  • Sustained Pharmacological beta IIPKC Inhibition Is Cardioprotective In Late-stage Hypertrophy And End-stage Heart Failure In Two Rat Models 81st Annual Scientific Session of the American-Heart-Association Ferreira, J. C., Koyanagi, T., Inagaki, K., Fajardo, G., Churchill, E. N., Budas, G., Disatnik, M., Tsutsui, J. M., Kihara, Y., Bernstein, D., Brum, P. C., Mochly-Rosen, D. LIPPINCOTT WILLIAMS & WILKINS. 2008: S535?S535
  • Ischemic Preconditioning Regulates Myocardial Viability Through The Proteasomal Regulation Of Two Opposing PKC Isozymes 81st Annual Scientific Session of the American-Heart-Association Churchill, E., Ferreira, J., Brum, P., Szweda, L., Mochly-Rosen, D. LIPPINCOTT WILLIAMS & WILKINS. 2008: S294?S294
  • Intracellular mechanisms of specific beta-adrenoceptor antagonists involved in improved cardiac function and survival in a genetic model of heart failure JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY Bartholomeu, J. B., Vanzelli, A. S., Rolim, N. P., Ferreira, J. C., Bechara, L. R., Tanaka, L. Y., Rosa, K. T., Alves, M. M., Medeiros, A., Mattos, K. C., Coelho, M. A., Irigoyen, M. C., Krieger, E. M., Krieger, J. E., Negrao, C. E., Ramires, P. R., Guatimosim, S., Brum, P. C. 2008; 45 (2): 240-249

    Abstract

    beta-blockers, as class, improve cardiac function and survival in heart failure (HF). However, the molecular mechanisms underlying these beneficial effects remain elusive. In the present study, metoprolol and carvedilol were used in doses that display comparable heart rate reduction to assess their beneficial effects in a genetic model of sympathetic hyperactivity-induced HF (alpha(2A)/alpha(2C)-ARKO mice). Five month-old HF mice were randomly assigned to receive either saline, metoprolol or carvedilol for 8 weeks and age-matched wild-type mice (WT) were used as controls. HF mice displayed baseline tachycardia, systolic dysfunction evaluated by echocardiography, 50% mortality rate, increased cardiac myocyte width (50%) and ventricular fibrosis (3-fold) compared with WT. All these responses were significantly improved by both treatments. Cardiomyocytes from HF mice showed reduced peak [Ca(2+)](i) transient (13%) using confocal microscopy imaging. Interestingly, while metoprolol improved [Ca(2+)](i) transient, carvedilol had no effect on peak [Ca(2+)](i) transient but also increased [Ca(2+)] transient decay dynamics. We then examined the influence of carvedilol in cardiac oxidative stress as an alternative target to explain its beneficial effects. Indeed, HF mice showed 10-fold decrease in cardiac reduced/oxidized glutathione ratio compared with WT, which was significantly improved only by carvedilol treatment. Taken together, we provide direct evidence that the beneficial effects of metoprolol were mainly associated with improved cardiac Ca(2+) transients and the net balance of cardiac Ca(2+) handling proteins while carvedilol preferentially improved cardiac redox state.

    View details for DOI 10.1016/j.yjmcc.2008.05.011

    View details for Web of Science ID 000258853400016

    View details for PubMedID 18632114

  • The role of local and systemic renin angiotensin system activation in a genetic model of sympathetic hyperactivity-induced heart failure in mice AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Ferreira, J. C., Bacurau, A. V., Evangelista, F. S., Coelho, M. A., Oliveira, E. M., Casarini, D. E., Krieger, J. E., Brum, P. C. 2008; 294 (1): R26-R32

    Abstract

    Sympathetic hyperactivity (SH) and renin angiotensin system (RAS) activation are commonly associated with heart failure (HF), even though the relative contribution of these factors to the cardiac derangement is less understood. The role of SH on RAS components and its consequences for the HF were investigated in mice lacking alpha(2A) and alpha(2C) adrenoceptor knockout (alpha(2A)/alpha(2C)ARKO) that present SH with evidence of HF by 7 mo of age. Cardiac and systemic RAS components and plasma norepinephrine (PN) levels were evaluated in male adult mice at 3 and 7 mo of age. In addition, cardiac morphometric analysis, collagen content, exercise tolerance, and hemodynamic assessments were made. At 3 mo, alpha(2A)/alpha(2C)ARKO mice showed no signs of HF, while displaying elevated PN, activation of local and systemic RAS components, and increased cardiomyocyte width (16%) compared with wild-type mice (WT). In contrast, at 7 mo, alpha(2A)/alpha(2C)ARKO mice presented clear signs of HF accompanied only by cardiac activation of angiotensinogen and ANG II levels and increased collagen content (twofold). Consistent with this local activation of RAS, 8 wk of ANG II AT(1) receptor blocker treatment restored cardiac structure and function comparable to the WT. Collectively, these data provide direct evidence that cardiac RAS activation plays a major role underlying the structural and functional abnormalities associated with a genetic SH-induced HF in mice.

    View details for DOI 10.1152/ajpregu.00424.2007

    View details for Web of Science ID 000252243800004

    View details for PubMedID 17977919

  • Maximal lactate steady state in running mice: Effect of exercise training CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY Ferreira, J. C., Rolim, N. P., Bartholomeu, J. B., Gobatto, C. A., Kokubun, E., Brum, P. C. 2007; 34 (8): 760-765

    Abstract

    1. Maximal lactate steady state (MLSS) corresponds to the highest blood lactate concentration (MLSSc) and workload (MLSSw) that can be maintained over time without continual blood lactate accumulation and is considered an important marker of endurance exercise capacity. The present study was undertaken to determine MLSSw and MLSSc in running mice. In addition, we provide an exercise training protocol for mice based on MLSSw. 2. Maximal lactate steady state was determined by blood sampling during multiple sessions of constant-load exercise varying from 9 to 21 m/min in adult male C57BL/6J mice. The constant-load test lasted at least 21 min. The blood lactate concentration was analysed at rest and then at 7 min intervals during exercise. 3. The MLSSw was found to be 15.1 +/- 0.7 m/min and corresponded to 60 +/- 2% of maximal speed achieved during the incremental exercise testing. Intra- and interobserver variability of MLSSc showed reproducible findings. Exercise training was performed at MLSSw over a period of 8 weeks for 1 h/day and 5 days/week. Exercise training led to resting bradycardia (21%) and increased running performance (28%). Of interest, the MLSSw of trained mice was significantly higher than that in sedentary littermates (19.0 +/- 0.5 vs 14.2 +/- 0.5 m/min; P = 0.05), whereas MLSSc remained unchanged (3.0 mmol/L). 4. Altogether, we provide a valid and reliable protocol to improve endurance exercise capacity in mice performed at highest workload with predominant aerobic metabolism based on MLSS assessment.

    View details for DOI 10.1111/j.1440-1681.2007.04635.x

    View details for Web of Science ID 000247575500011

    View details for PubMedID 17600553

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