Rationale: C2238 atrial natriuretic peptide (ANP) minor allele (substitution of thymidine with cytosine in position 2238) associates with increased risk of cardiovascular events. Objective: We investigated the mechanisms underlying the vascular effects of C2238-alpha ANP. Methods and Results: In vitro, human umbilical vein endothelial cell were exposed to either wild-type (T2238)- or mutant (C2238)-alpha ANP. Cell survival and apoptosis were tested by Trypan blue, annexin V, and cleaved caspase-3 assays. C2238-alpha ANP significantly reduced human umbilical vein endothelial cell survival and increased apoptosis. In addition, C2238-alpha ANP reduced endothelial tube formation, as assessed by matrigel. C2238-alpha ANP did not differentially modulate natriuretic peptide receptor (NPR)-A/B activity with respect to T2238-alpha ANP, as evaluated by intracellular cGMP levels. In contrast, C2238-alpha ANP, but not T2238-alpha ANP, markedly reduced intracellular cAMP levels in an NPR-C-dependent manner. Accordingly, C2238-alpha ANP showed higher affinity binding to NPR-C, than T2238-alpha ANP. Either NPR-C inhibition by antisense oligonucleotide or NPR-C gene silencing by small interfering RNA rescued survival and tube formation of human umbilical vein endothelial cell exposed to C2238-alpha ANP. Similar data were obtained in human aortic endothelial cell with NPR-C knockdown. NPR-C activation by C2238-alpha ANP inhibited the protein kinase A/Akt1 pathway and increased reactive oxygen species. Adenovirus-mediated Akt1 reactivation rescued the detrimental effects of C2238-alpha ANP. Overall, these data indicate that C2238-alpha ANP affects endothelial cell integrity through NPR-C-dependent inhibition of the cAMP/protein kinase A/Akt1 pathway and increased reactive oxygen species production. Accordingly, C2238-alpha ANP caused impairment of acetylcholine-dependent vasorelaxation ex vivo, which was rescued by NPR-C pharmacological inhibition. Finally, subjects carrying C2238 minor allele showed early endothelial dysfunction, which highlights the clinical relevance of our results. Conclusions: C2238-alpha ANP reduces endothelial cell survival and impairs endothelial function through NPR-C signaling. NPR-C targeting represents a potential strategy to reduce cardiovascular risk in C2238 minor-allele carriers.

C2238 Atrial Natriuretic Peptide Molecular Variant Is Associated With Endothelial Damage and Dysfunction Through Natriuretic Peptide Receptor C Signaling / Sciarretta, Sebastiano; S. S., Marchitti; F., Bianchi; A., Moyes; E., Barbato; S., Di Castro; R., Stanzione; M., Cotugno; L., Castello; Calvieri, Camilla; I., Eberini; J., Sadoshima; A. J., Hobbs; Volpe, Massimo; Rubattu, Speranza Donatella. - In: CIRCULATION RESEARCH. - ISSN 0009-7330. - STAMPA. - 112:10(2013), pp. 1355-1364. [10.1161/circresaha.113.301325]

C2238 Atrial Natriuretic Peptide Molecular Variant Is Associated With Endothelial Damage and Dysfunction Through Natriuretic Peptide Receptor C Signaling

SCIARRETTA, SEBASTIANO;E. Barbato;CALVIERI, CAMILLA;VOLPE, Massimo;RUBATTU, Speranza Donatella
2013

Abstract

Rationale: C2238 atrial natriuretic peptide (ANP) minor allele (substitution of thymidine with cytosine in position 2238) associates with increased risk of cardiovascular events. Objective: We investigated the mechanisms underlying the vascular effects of C2238-alpha ANP. Methods and Results: In vitro, human umbilical vein endothelial cell were exposed to either wild-type (T2238)- or mutant (C2238)-alpha ANP. Cell survival and apoptosis were tested by Trypan blue, annexin V, and cleaved caspase-3 assays. C2238-alpha ANP significantly reduced human umbilical vein endothelial cell survival and increased apoptosis. In addition, C2238-alpha ANP reduced endothelial tube formation, as assessed by matrigel. C2238-alpha ANP did not differentially modulate natriuretic peptide receptor (NPR)-A/B activity with respect to T2238-alpha ANP, as evaluated by intracellular cGMP levels. In contrast, C2238-alpha ANP, but not T2238-alpha ANP, markedly reduced intracellular cAMP levels in an NPR-C-dependent manner. Accordingly, C2238-alpha ANP showed higher affinity binding to NPR-C, than T2238-alpha ANP. Either NPR-C inhibition by antisense oligonucleotide or NPR-C gene silencing by small interfering RNA rescued survival and tube formation of human umbilical vein endothelial cell exposed to C2238-alpha ANP. Similar data were obtained in human aortic endothelial cell with NPR-C knockdown. NPR-C activation by C2238-alpha ANP inhibited the protein kinase A/Akt1 pathway and increased reactive oxygen species. Adenovirus-mediated Akt1 reactivation rescued the detrimental effects of C2238-alpha ANP. Overall, these data indicate that C2238-alpha ANP affects endothelial cell integrity through NPR-C-dependent inhibition of the cAMP/protein kinase A/Akt1 pathway and increased reactive oxygen species production. Accordingly, C2238-alpha ANP caused impairment of acetylcholine-dependent vasorelaxation ex vivo, which was rescued by NPR-C pharmacological inhibition. Finally, subjects carrying C2238 minor allele showed early endothelial dysfunction, which highlights the clinical relevance of our results. Conclusions: C2238-alpha ANP reduces endothelial cell survival and impairs endothelial function through NPR-C signaling. NPR-C targeting represents a potential strategy to reduce cardiovascular risk in C2238 minor-allele carriers.
2013
natriuretic peptide receptor type c; akt; t2238c gene variant; endothelial dysfunction; atrial natriuretic peptide
01 Pubblicazione su rivista::01a Articolo in rivista
C2238 Atrial Natriuretic Peptide Molecular Variant Is Associated With Endothelial Damage and Dysfunction Through Natriuretic Peptide Receptor C Signaling / Sciarretta, Sebastiano; S. S., Marchitti; F., Bianchi; A., Moyes; E., Barbato; S., Di Castro; R., Stanzione; M., Cotugno; L., Castello; Calvieri, Camilla; I., Eberini; J., Sadoshima; A. J., Hobbs; Volpe, Massimo; Rubattu, Speranza Donatella. - In: CIRCULATION RESEARCH. - ISSN 0009-7330. - STAMPA. - 112:10(2013), pp. 1355-1364. [10.1161/circresaha.113.301325]
File allegati a questo prodotto
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/514762
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? 21
  • Scopus 34
  • ???jsp.display-item.citation.isi??? 33
social impact