Abstract
The feasibility of gene therapy for cardiomyopathy, heart failure and other chronic cardiac muscle diseases is so far unproven. Here, we developed an in vivo recombinant adeno-associated virus (rAAV) transcoronary delivery system that allows stable, high efficiency and relatively cardiac-selective gene expression. We used rAAV to express a pseudophosphorylated mutant of human phospholamban (PLN), a key regulator of cardiac sarcoplasmic reticulum (SR) Ca2+ cycling in BIO14.6 cardiomyopathic hamsters. The rAAV/S16EPLN treatment enhanced myocardial SR Ca2+ uptake and suppressed progressive impairment of left ventricular (LV) systolic function and contractility for 28–30 weeks, thereby protecting cardiac myocytes from cytopathic plasma-membrane disruption. Low LV systolic pressure and deterioration in LV relaxation were also largely prevented by rAAV/S16EPLN treatment. Thus, transcoronary gene transfer of S16EPLN via rAAV vector is a potential therapy for progressive dilated cardiomyopathy and associated heart failure.
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References
Chien, K.R. Stress pathways and heart failure. Cell 98, 555–558 (1999).
Hunter, J.J. & Chien, K.R. Signaling pathways for cardiac hypertrophy and failure. N. Engl. J. Med. 341, 1276–1283 (1999).
Hoshijima, M. & Chien, K.R. Mixed signals in heart failure: cancer rules. J. Clin. Invest. 109, 849–855 (2002).
Minamisawa, S. et al. Chronic phospholamban-sarcoplasmic reticulum calcium ATPase interaction is the critical calcium cycling defect in dilated cardiomyopathy. Cell 99, 313–322 (1999).
Arber, S. et al. MLP-deficient mice exhibit a disruption of cardiac cytoarchitectural organization, dilated cardiomyopathy, and heart failure. Cell 88, 393–403 (1997).
Sato, Y. et al. Rescue of contractile parameters and myocyte hypertrophy in calsequestrin overexpressing myocardium by phospholamban ablation. J. Biol. Chem. 276, 9392–9399 (2001).
Freeman, K. et al. Alterations in cardiac adrenergic signaling and calcium cycling differentially affect the progression of cardiomyopathy. J. Clin. Invest. 107, 967–74 (2001).
Miyamoto, M.I. et al. Adenoviral gene transfer of SERCA2a improves left-ventricular function in aortic-banded rats in transition to heart failure. Proc. Natl. Acad. Sci. USA 97, 793–798 (2000).
del Monte, F. et al. Improvement in survival and cardiac metabolism after gene transfer of sarcoplasmic reticulum Ca(2+)-ATPase in a rat model of heart failure. Circulation 104, 1424–1429 (2001).
White, D.C. et al. Preservation of myocardial beta-adrenergic receptor signaling delays the development of heart failure after myocardial infarction. Proc. Natl. Acad. Sci USA 97, 5428–5433 (2000).
Shah, A.S. et al. In vivo ventricular gene delivery of a beta-adrenergic receptor kinase inhibitor to the failing heart reverses cardiac dysfunction. Circulation 103, 1311–1316 (2001).
Ikeda, Y. et al. Altered membrane proteins and permeability correlate with cardiac dysfunction in cardiomyopathic hamsters. Am. J. Physiol. Heart Circ. Physiol. 278, H1362–1370 (2000).
Straub, V. & Campbell, K.P. Muscular dystrophies and the dystrophin-glycoprotein complex. Curr. Opin. Neurol. 10, 168–75 (1997).
Ikeda, Y. et al. Restoration of deficient membrane proteins in the cardiomyopathic hamster by in vivo cardiac gene transfer. Circulation 105, 502–8 (2002).
Tada, M. & Toyofuku, T. Cardiac sarcoplasmic reticulum Ca2+-ATPase. in Handbook of Physiology, Section 2, The Cardiovascular system, Volume 1: The Heart ed. Berne, R.M. 301–334 (Oxford University Press, 2001).
Simmerman, H.K. & Jones, L.R. Phospholamban: Protein structure, mechanism of action, and role in cardiac function. Physiol. Rev. 78, 921–947 (1998).
Antos, C.L. et al. Dilated cardiomyopathy and sudden death resulting from constitutive activation of protein kinase a. Circ. Res. 89, 997–1004 (2001).
Ryoke, T. et al. Progressive cardiac dysfunction and fibrosis in the cardiomyopathic hamster and effects of growth hormone and angiotensin-converting enzyme inhibition. Circulation 100, 1734–1743 (1999).
Lai, N.C. et al. Intracoronary delivery of adenovirus encoding adenylyl cyclase VI increases left ventricular function and cAMP-generating capacity. Circulation 102, 2396–2401 (2000).
Logeart, D. et al. Highly efficient adenovirus-mediated gene transfer to cardiac myocytes after single-pass coronary delivery. Hum. Gene Ther. 11, 1015–1022 (2000).
Brauner, R. et al. Intracoronary adenovirus-mediated transfer of immunosuppressive cytokine genes prolongs allograft survival. J. Thorac. Cardiovasc. Surg. 114, 923–933 (1997).
Svensson, E.C. et al. Efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors. Circulation 99, 201–205 (1999).
Kawada, T. et al. Rescue of hereditary form of dilated cardiomyopathy by rAAV-mediated somatic gene therapy: Amelioration of morphological findings, sarcolemmal permeability, cardiac performances, and the prognosis of TO- 2 hamsters. Proc. Natl. Acad. Sci. USA 99, 901–906 (2002).
Melo, L.G. et al. Gene therapy strategy for long-term myocardial protection using adeno- associated virus-mediated delivery of heme oxygenase gene. Circulation 105, 602–607 (2002).
Xiao, X., Li, J. & Samulski, R.J. Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector. J. Virol. 70, 8098–8108 (1996).
Kessler, P.D. et al. Gene delivery to skeletal muscle results in sustained expression and systemic delivery of a therapeutic protein. Proc. Natl. Acad. Sci. USA 93, 14082–14087 (1996).
Kay, M.A. et al. Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector. Nature Genet. 24, 257–261 (2000).
Summerford, C. & Samulski, R.J. Membrane-associated heparan sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J. Virol. 72, 1438–1445 (1998).
Pruchnic, R. et al. The use of adeno-associated virus to circumvent the maturation- dependent viral transduction of muscle fibers. Hum. Gene Ther. 11, 521–536 (2000).
Asundi, V.K., Keister, B.F., Stahl, R.C. & Carey, D.J. Developmental and cell-type-specific expression of cell surface heparan sulfate proteoglycans in the rat heart. Exp. Cell Res. 230, 145–153 (1997).
Snyder, R.O. et al. Persistent and therapeutic concentrations of human factor IX in mice after hepatic gene transfer of recombinant AAV vectors. Nature Genet. 16, 270–276 (1997).
Nakai, H. et al. Adeno-associated viral vector-mediated gene transfer of human blood coagulation factor IX into mouse liver. Blood 91, 4600–4607 (1998).
Coral-Vazquez, R. et al. Disruption of the sarcoglycan-sarcospan complex in vascular smooth muscle: a novel mechanism for cardiomyopathy and muscular dystrophy. Cell 98, 465–474 (1999).
Xiao, X., Li, J. & Samulski, R.J. Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J. Virol. 72, 2224–2232 (1998).
Auricchio, A., Hildinger, M., O'Connor, E., Gao, G.P. & Wilson, J.M. Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. Hum. Gene Ther. 12, 71–76 (2001).
Frank, K., Tilgmann, C., Shannon, T.R., Bers, D.M. & Kranias, E.G. Regulatory role of phospholamban in the efficiency of cardiac sarcoplasmic reticulum Ca2+ transport. Biochemistry 39, 14176–14182 (2000).
Jackson, W.A. & Colyer, J. Translation of Ser16 and Thr17 phosphorylation of phospholamban into Ca2+-pump stimulation. Biochem. J. 316, 201–207 (1996).
Christensen, G., Minamisawa, S., Gruber, P.J., Wang, Y. & Chien, K.R. High-efficiency, long-term cardiac expression of foreign genes in living mouse embryos and neonates. Circulation 101, 178–184 (2000).
Acknowledgements
We thank J. Chrast and J. Lam for experimental assistance. This work was entirely supported by the Jean Le Ducq Foundation.
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K.R.C., M.H. and Y.W. have equity in a startup company, Celladon, that holds the intellectual property for the mutant phospholamban (PLN) gene therapy strategy including S16EPLN outlined in this paper.
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Hoshijima, M., Ikeda, Y., Iwanaga, Y. et al. Chronic suppression of heart-failure progression by a pseudophosphorylated mutant of phospholamban via in vivo cardiac rAAV gene delivery. Nat Med 8, 864–871 (2002). https://doi.org/10.1038/nm739
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DOI: https://doi.org/10.1038/nm739
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