Percutaneous Cardiac Recirculation-Mediated Gene Transfer of an Inhibitory Phospholamban Peptide Reverses Advanced Heart Failure in Large Animals
David M. Kaye, MD, PhD*,1,2,*,
Arthur Preovolos, BS*,1,3,
Tanneale Marshall, BS*,
Melissa Byrne, PhD*,1,3,
Masahiko Hoshijima, PhD ,
Roger Hajjar, MD ,
Justin A. Mariani, MD*,
Salvatore Pepe, PhD*,
Kenneth R. Chien, MD, PhD and
John M. Power, PhD*,1,2
* Baker Heart Research Institute, Melbourne, Australia
Institute of Molecular Medicine, University of California–San Diego, La Jolla, California
Massachusetts General Hospital, Cardiovascular Research Center, Harvard Medical School, and the Harvard Stem Cell Institute, Boston, Massachusetts.
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Figure 1 Percutaneous Approach to Recirculating Cardiac Gene Delivery
(A) Diagram demonstrates the position of catheters placed within the heart. (B) Circuit diagram for recirculating gene delivery to the myocardium. V = viral vector.
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Figure 2 Perfusate Distribution Pattern Following Cardiac Recirculation
Pattern of distribution of indocyanine green through the myocardium during recirculation, as determined by near-infrared spectroscopy (left panels = anterior view; right panels = cross section). Images show a clear demarcation between perfused and nonperfused regions, the latter corresponding with the nonperfused right coronary artery distribution.
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Figure 3 Myocardial Pattern of Gene Expression Following Recirculating Delivery
(A) Hematoxylin and eosin-stained section of nonfailing myocardium 2 weeks after cardiac adenovirus ß-galactosidase (AdLacZ) recirculation, without evidence of inflammation or ischemic damage. (B and C) Immunohistochemical sections of nonfailing myocardium stained for ß-galactosidase 2 weeks after direct intracoronary (B) and recirculating (C) delivery of AdLacZ.
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Figure 4 Delivery Mode Determines the Extent of Extracardiac Transgene Expression
Immunohistochemical sections of lung stained for ß-galactosidase 2 weeks after direct intracoronary (A) and recirculating (B) delivery of adenovirus ß-galactosidase (AdLacZ). Immunohistochemical sections of liver stained for ß-galactosidase after direct intracoronary (C) and recirculating (D) delivery of AdLacZ.
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Figure 5 Hemodynamic Effect of S16E Gene Delivery on the Failing Heart
Echocardiographic and hemodynamic effects of AdS16EPLN (n = 9) gene transfer to the failing ovine heart compared with AdLacZ (lacZ) (n = 6) treated animals. (A) Left ventricular (LV) ejection fraction; (B) LV end-diastolic area;(C) LV end-diastolic pressure (EDP); (D) Peak positive dP/dt. Solid bars = before gene delivery; open bars = 2 weeks after gene delivery. AdLacZ = adenovirus ß-galactosidase; AdS16EPLN = adenovirus serine-to-glutamate "pseudo-phosphorylated" phospholamban mutant.
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Figure 6 Effect of S16E Gene Delivery on Phospholamban Expression
(A) Western blots showing expression of total and phospho-phospholamban (pPLN) in nonfailing and failing myocardium (after AdLacZ and AdS16EPLN). (B) Bar graphs demonstrating the pPLN:total phospholamban (tPLN) ratio in control nonfailing (n = 4) and heart failure (HF) AdLacZ (n = 4) and AdS16E (n = 5) treated animals. *p = 0.05 versus control;  p < 0.05 versus HF-AdLacZ. Abbreviations as in Figure 5.
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Figure 7 Effect of S16E Gene Delivery on SERCA Activity
Sarcoplasmic reticulum Ca2+-adenosine triphosphatase (ATPase) (SERCA) activity in control nonfailing (n = 3) and heart failure (HF) AdLacZ (n = 4) and AdS16E (n = 4) treated animals. Abbreviations as in Figure 5.
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