Metabolic and functional consequences of cytosolic  5′-nucleotidase-IA overexpression in neonatal rat cardiomyocytes

Sala-Newby, Graciela B.; Freeman, Norman H; Curto, Maria de Los Angeles; Newby, Andrew C.

doi:10.1152/ajpheart.00053.2003

Cited by 15 publications

(18 citation statements)

References 40 publications

(32 reference statements)

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“…RNCMs were isolated from 2-to 3-day-old Wistar rats (bred at the University of Bristol), as previously described. 31 Cardiomyocytes were seeded on gelatin-coated plates and maintained in 4 : 1 DMEM-M199 serum-free medium. The level of purity of RNCM cultures was routinely assessed by staining for the cardiac marker a-sarcomeric actin.…”

Section: Discussionmentioning

confidence: 99%

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Identification of the prosurvival activity of nerve growth factor on cardiac myocytes

et al. 2007

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Neurotrophins (NTs) control neuron survival and regeneration. Recent research showed that NTs possess cardiovascular actions. In this study, we investigated the hypothesis that the NT nerve growth factor (NGF) prevents cardiomyocyte apoptosis. We demonstrated that cultured rat neonatal cardiomyocytes (RNCMs) produce NGF and express its trkA (tropomyosin-related receptor A (NGF high-affinity receptor)) receptor. RNCMs given a neutralizing antibody for NGF or the trkA inhibitor K252a underwent apoptosis, thus suggesting that NGF is an endogenous prosurvival factor for cardiomyocytes. Adenovirus (Ad)-mediated NGF overexpression protected RNCMs from apoptosis induced by either hypoxia/reoxygenation or angiotensin II (AngII). Similarly, recombinant NGF inhibited AngII-induced apoptosis in isolated rat adult cardiomyocytes. Finally, in a rat model of myocardial infarction, NGF gene transfer promoted cardiomyocyte survival. In RNCMs, recombinant NGF induced trkA phosphorylation, followed by Ser473 phosphorylation and nuclear translocation of phospho-protein kinase B (Akt). In response to Akt activation, Forkhead transcription factors Foxo-3a and Foxo-1 were phosphorylated and excluded from the nucleus. The prosurvival effect of adenoviral vector carrying the human NGF gene was inhibited in vitro by K252a, LY294002 (a panphosphatidyl inositol 3-kinase -PI3K -inhibitor), an Akt small interfering RNA, and adenoviruses carrying a dominant negative mutant form of Akt (Ad.DN.Akt) or an Akt-resistant Foxo-3a (Ad. . These results newly demonstrate the cardiac prosurvival action of NGF and provide mechanistic information on the signaling pathway, which encompasses trkA, PI3K-Akt, and Foxo. Neurotrophins (NTs) were initially considered for their prosurvival and regenerative actions on the nervous system. These beneficial actions of NTs are mediated by their tropomyosin-related kinase receptors (trks), which are tyrosin kinases.1,2 Recent studies have unraveled the potential of nerve growth factor (NGF) and other NTs for vascular biology.3-7 Our group discovered that NGF protects endothelial cells and skeletal myocytes from apoptosis and promotes neovascularization in murine ischemic limb muscles and diabetic skin wounds. 3,5,6 We demonstrated the involvement of protein kinase B (Akt) in NGF-induced angiogenesis. 3Although others already tracked the presence of NGF in the heart and isolated cardiomyocytes, [8][9][10][11] the hypothesis that NGF controls cardiomyocyte survival was, to the best of our knowledge, never investigated.In noncardiovascular cells, NGF, via Akt, promotes the nuclear exclusion of Foxo-3a and Foxo-1, two members of the Forkhead box, class O (Foxo) subfamily of Forkhead transcription factors, which stimulate cell death.12 Nuclear localization of Foxo activates several Foxo-responsive genes, including the proapoptotic TNFa, Fas ligand, and Bim. [13][14][15][16] Akt-dependent Foxo phosphorylation inactivates the Foxo nuclear translocation signal and promotes the interaction of Foxo with 14-3-3 proteins, thus k...

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Section: Discussionmentioning

confidence: 99%

“…Viral stocks were amplified, CsCl banded, and titrated. 31 NGF gene transfer to cultured RNCMs. RNCMs were grown in 4:1 DMEM-M199 and were infected overnight with Ad.hNGF or Ad.bGal (each at 50 M.O.I.).…”

Section: Discussionmentioning

confidence: 99%

Identification of the prosurvival activity of nerve growth factor on cardiac myocytes

et al. 2007

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“…Viral stocks were amplified, CsCl banded, and titrated as previously described. 41 Cells were plated 24 hours before infection, incubated with fresh media containing the required multiplicity of infection (MOI) per cell of virus, left for 6 hours, washed, and maintained until harvesting.…”

Section: Methodsmentioning

confidence: 99%

Local Inhibition of MicroRNA-24 Improves Reparative Angiogenesis and Left Ventricle Remodeling and Function in Mice With Myocardial Infarction

et al. 2013

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Myocardial infarction (MI) is the leading cause of death worldwide. MicroRNAs regulate the expression of their target genes, thus mediating a plethora of pathophysiological functions. Recently, miRNA-24 emerged as an important but controversial miRNA involved in post-MI responses. Here, we aimed at clarifying the effect of adenovirus-mediate intra-myocardial delivery of a decoy for miRNA-24 in a mouse MI model and to investigate the impact of miRNA-24 inhibition on angiogenesis and cardiovascular apoptosis. After MI induction, miRNA-24 expression was lower in the peri-infarct tissue and its resident cardiomyocytes and fibroblasts; while it increased in endothelial cells (ECs). Local adenovirus-mediated miRNA-24 decoy delivery increased angiogenesis and blood perfusion in the peri-infarct myocardium, reduced infarct size, induced fibroblast apopotosis and overall improved cardiac function. Notwithstanding these beneficial effects, miRNA-24 decoy increased cardiomyocytes apoptosis. In vitro, miRNA-24 inhibition enhanced ECs survival, proliferation and networking in capillary-like tubes and induced cardiomyocyte and fibroblast apoptosis. Finally, we identified eNOS as a novel direct target of miR-24 in human cultured ECs and in vivo. Our findings suggest that miRNA-24 inhibition exerts distinct biological effects on ECs, cardiomyocytes and fibroblasts. The overall result of post-infarction local miRNA-24 inhibition appears to be therapeutic.

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“…We used two adenoviral vectors prepared at the Bristol Heart Institute (Bristol Royal Infirmary, University of Bristol, United Kingdom) as previously described [25]: a control vector containing a GFP cDNA as tag (Ad-GFP vector) and a vector containing the coding sequence of pigeon cN-IA (5′-nucleotidase) without GFP cDNA (Ad-cN-IA vector). Virus stocks were amplified, CsCl banded and titrated before use.…”

Section: Methodsmentioning

confidence: 99%

Effects of Adenosine on Lymphangiogenesis

et al. 2014

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BackgroundThe lymphatic system controls tissue homeostasis by draining protein-rich lymph to the vascular system. Lymphangiogenesis, the formation of lymphatic vessels, is a normal event in childhood but promotes tumor spread and metastasis during adulthood. Blocking lymphangiogenesis may therefore be of therapeutic interest. Production of adenosine is enhanced in the tumor environment and contributes to tumor progression through stimulation of angiogenesis. In this study, we determined whether adenosine affects lymphangiogenesis.MethodsLymphatic endothelial cells (HMVEC-dLy) were cultured in presence of adenosine and their proliferation, migration and tube formation was assessed. Gelatin sponges embedded with the stable analogue of adenosine 2-chloro adenosine were implanted in mice ear and lymphangiogenesis was quantified. Mice were intravenously injected with adenoviruses containing expression vector for 5′-endonucleotidase, which plays a major role in the formation of adenosine.ResultsIn vitro, we observed that adenosine decreased the proliferation of lymphatic endothelial cells, their migration and tube formation. However, in vivo, gelatin sponges containing 2-chloro adenosine and implanted in mice ear displayed an elevated level of lymphangiogenesis (2.5-fold, p<0.001). Adenovirus-mediated over-expression of cytosolic 5′-nucleotidase IA stimulated lymphangiogenesis and the recruitment of macrophages in mouse liver. Proliferation of lymphatic endothelial cells was enhanced (2-fold, p<0.001) when incubated in the presence of conditioned medium from murine macrophages.ConclusionWe have shown that adenosine stimulates lymphangiogenesis in vivo, presumably through a macrophage-mediated mechanism. This observation suggests that blockade of adenosine receptors may help in anti-cancer therapies.

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Metabolic and functional consequences of cytosolic 5′-nucleotidase-IA overexpression in neonatal rat cardiomyocytes

Cited by 15 publications

References 40 publications

Identification of the prosurvival activity of nerve growth factor on cardiac myocytes

Identification of the prosurvival activity of nerve growth factor on cardiac myocytes

Local Inhibition of MicroRNA-24 Improves Reparative Angiogenesis and Left Ventricle Remodeling and Function in Mice With Myocardial Infarction

Effects of Adenosine on Lymphangiogenesis

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