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(Circulation. 2002;106:I-125.)
© 2002 American Heart Association, Inc.

Thoracic Transplantation and Mechanical Support for Congestive Heart Failure

Cellular Cardiomyoplasty of Cardiac Fibroblasts by Adenoviral Delivery of MyoD Ex Vivo: An Unlimited Source of Cells for Myocardial Repair

Sharon Etzion, MSc; Israel M. Barbash, BmedSc; Micha S. Feinberg, MD; Parvin Zarin, BSc; Liron Miller, BSc; Esther Guetta, PhD; Radka Holbova; Robert A. Kloner, MD, PhD; Laurence H. Kedes, MD; Jonathan Leor, MD

From the Neufeld Cardiac Research Institute, Tel-Aviv University, Sheba Medical Center, Tel-Hashomer, Israel (S.E., I.B., M.F., L.M., E.G., R.H., J.L.); Cardiac Research Center, Faculty of Health Sciences, Ben-Gurion University, Beer-Sheva, Israel (S.E., P.Z.); Heart Institute, Good Samaritan Hospital, Los Angeles, Calif. (R.K.); Department of Medicine, Keck School of Medicine of the University of Southern California, Los Angeles, Calif. (R.K., L.K.); and the Institute for Genetic Medicine, Department of Biochemistry & Molecular Biology, Los Angeles, Calif. (L.K.).

Correspondence to Jonathan Leor, MD, Neufeld Cardiac Research Institute, Sheba Medical Center, Tel-Hashomer 52621, Israel. E-mail leorj{at}post.tau.ac.il

Abstract

Background The muscle-specific MyoD family of transcription factors function as master genes that are able to prompt myogenesis in a variety of cells. The purpose of our study was to determine whether MyoD could induce primary cardiac fibroblasts, isolated from infarcted myocardium or pericardium, to undergo myogenic conversion in a clinically relevant approach.

Methods and Results Primary rat fibroblasts from 7-day-old infarcted myocardium or normal pericardium were transfected by an E1/E3-deleted adenoviral vector carrying both a human MyoD cDNA driven by a CMV promoter and a green fluorescent protein (GFP) reporter gene driven by a second CMV promoter. Expression of MyoD caused myogenic differentiation of cultured fibroblasts, as defined by elongation and fusion into multinucleated myotubes, typical cross striation as identified by electron microscopy, and positive immunostaining for sarcomeric actin, fast myosin heavy chain (MHC), and actinin. The myogenic cells (1.5x10⁶) were transplanted into the infarcted myocardium 7 days after coronary artery occlusion. By 1 month after transplantation, the converted fibroblasts gave rise to a cluster of myogenic cells that in a few hearts occupied a large part of the scar with positive immunostaining for the myogenic proteins fast-MHC and sarcomeric actin. A few cells expressed the gap junction protein connexin 43 in a disorganized manner. There was no positive staining in the control hearts treated with injections of untreated fibroblasts or culture medium.

Conclusions Our work shows that it is possible to exploit the unique capacity of MyoD to activate myogenesis in fibroblasts ex vivo and to create a vast source of autologous myogenic cells for transplantation.

Key Words: cells • gene therapy • myocytes • transplantation