Restoration of beta-adrenergic receptor signaling and contractile function in heart failure by disruption of the betaARK1/phosphoinositide 3-kinase complex.
BACKGROUND
Desensitization and downregulation of myocardial beta-adrenergic receptors (betaARs) are initiated by the increase in betaAR kinase 1 (betaARK1) levels. By interacting with betaARK1 through the phosphoinositide kinase (PIK) domain, phosphoinositide 3-kinase (PI3K) is targeted to agonist-stimulated betaARs, where it regulates endocytosis. We tested the hypothesis that inhibition of receptor-targeted PI3K activity would alter receptor trafficking and ameliorate betaAR signaling, ultimately improving contractility of failing cardiomyocytes.


RESULTS
To competitively displace PI3K from betaARK1, we generated mice with cardiac-specific overexpression of the PIK domain. Seven-day isoproterenol administration in wild-type mice induced desensitization of betaARs and their redistribution from the plasma membrane to early and late endosomes. In contrast, transgenic PIK overexpression prevented the redistribution of betaARs away from the plasma membrane and preserved their responsiveness to agonist. We further tested whether PIK overexpression could normalize already established betaAR abnormalities and ameliorate contractile dysfunction in a large animal model of heart failure induced by rapid ventricular pacing in pigs. Failing porcine hearts showed increased betaARK1-associated PI3K activity and marked desensitization and redistribution of betaARs to endosomal compartments. Importantly, adenoviral gene transfer of the PIK domain in failing pig myocytes resulted in reduced receptor-localized PI3K activity and restored to nearly normal agonist-stimulated cardiomyocyte contractility.


CONCLUSIONS
These data indicate that the heart failure state is associated with a maladaptive redistribution of betaARs away from the plasma membrane that can be counteracted through a strategy that targets the betaARK1/PI3K complex.
Perrino
Cinzia
C
Department of Medicine, Cell Biology and Molecular Genetics, Duke University Medical Center, Durham, NC 27710, USA.
Naga Prasad
Sathyamangla V
SV
Schroder
Jacob N
JN
Hata
Jonathan A
JA
Milano
Carmelo
C
Rockman
Howard A
HA
D000818:Animals; D058534:Class I Phosphatidylinositol 3-Kinases; D011992:Endosomes / Q000378:metabolism; D015316:Genetic Therapy; D006333:Heart Failure / Q000628:therapy*; D007545:Isoproterenol / Q000008:administration & dosage / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D009200:Myocardial Contraction / Q000187:drug effects*; D019869:Phosphatidylinositol 3-Kinases / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport; D011943:Receptors, Adrenergic, beta / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D051552:beta-Adrenergic Receptor Kinases / Q000187:drug effects / Q000378:metabolism*
D011943:Receptors, Adrenergic, beta; D019869:Phosphatidylinositol 3-Kinases; D058534:Class I Phosphatidylinositol 3-Kinases; C484760:PIK3CA protein, human; D051552:beta-Adrenergic Receptor Kinases; D007545:Isoproterenol