SERCA2a gene transfer enhances eNOS expression and activity in endothelial cells.
Congestive heart failure (HF) is associated with impaired endothelium-dependent nitric oxide-mediated vasodilatation. The aim of this study was to examine the effects of sarco/endoplasmic reticulum (ER) Ca(2+)-ATPase 2a (SERCA2a) gene transfer on endothelial function in a swine HF model. Two months after the creation of mitral regurgitation to induce HF, the animals underwent intracoronary injection of adeno-associated virus (AAV) carrying SERCA2a (n = 7) or saline (n = 6). At 4 months, coronary flow (CF) was measured in the mid-portion of the left anterior descending (LAD) artery. In the failing animals, CF was decreased significantly; SERCA2a gene transfer rescued CF to levels observed in sham-group [ml/min/g, 0.47 +/- 0.064 saline versus 0.89 +/- 0.116, SERCA2a; P < 0.05; 1.00 +/- 0. 185 sham P = NS (nonsignificant)]. In coronary arteries from HF animals, SERCA2a and endothelial isoform of nitric oxide synthase (eNOS) protein expression were decreased, but restored to normal levels by SERCA2a gene transfer. In human coronary artery endothelial cells (HCAECs), SERCA2a overexpression increased eNOS expression, phosphorylation, eNOS promoter activity, Ca(2+) storage capacity, and enhanced histamine-induced calcium oscillations, eNOS activity, and cyclic guanosine monophosphate (cGMP) production. Thus, SERCA2a gene transfer increases eNOS expression and activity by modulating calcium homeostasis to improve CF. These findings suggest that SERCA2a gene transfer improves vascular reactivity in the setting of HF.
Hadri
Lahouaria
L
Cardiovascular Research Center, Mount Sinai School of Medicine, New York, New York 10029, USA.
Bobe
Regis
R
Kawase
Yoshiaki
Y
Ladage
Dennis
D
Ishikawa
Kiyotake
K
Atassi
Fabrice
F
Lebeche
Djamel
D
Kranias
Evangelia G
EG
Leopold
Jane A
JA
Lompré
Anne-Marie
AM
Lipskaia
Larissa
L
Hajjar
Roger J
RJ
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D003331:Coronary Vessels / Q000166:cytology; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D018014:Gene Transfer Techniques; D006333:Heart Failure / Q000378:metabolism / Q000628:therapy; D006801:Humans; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D013552:Swine
D052250:Nitric Oxide Synthase Type III; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases
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