Published online before print February 26, 2004, doi: 10.1161/01.RES.0000123827.60210.72
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© 2004 American Heart Association, Inc.
Integrative Physiology |
Human Mesenchymal Stem Cells as a Gene Delivery System to Create Cardiac Pacemakers
From the Institute of Molecular Cardiology, Departments of Physiology and Biophysics (I.P., Z.L., V.V., S.D., J.Z., J.G., Z.P., P.R.B., I.S.C.), SUNY Stony Brook, Stony Brook, NY; Center for Molecular Therapeutics, Department of Pharmacology (A.P., P.D., J.Q., I.N.S., R.B.R., M.R.R.), Department of Pediatrics (M.R.R.), and the Institute of Comparative Medicine and Department of Pathology (A.J.H.), Columbia University, New York, NY.
Correspondence to Michael R. Rosen, MD, Center for Molecular Therapeutics, Depts of Pharmacology and Pediatrics, Columbia University, 630 West 168 St, PH 7West-321, New York, NY 10032. E-mail mrr1{at}columbia.edu
We tested the ability of human mesenchymal stem cells (hMSCs) to deliver a biological pacemaker to the heart. hMSCs transfected with a cardiac pacemaker gene, mHCN2, by electroporation expressed high levels of Cs+-sensitive current (31.1±3.8 pA/pF at –150 mV) activating in the diastolic potential range with reversal potential of –37.5±1.0 mV, confirming the expressed current as If-like. The expressed current responded to isoproterenol with an 11-mV positive shift in activation. Acetylcholine had no direct effect, but in the presence of isoproterenol, shifted activation 15 mV negative. Transfected hMSCs influenced beating rate in vitro when plated onto a localized region of a coverslip and overlaid with neonatal rat ventricular myocytes. The coculture beating rate was 93±16 bpm when hMSCs were transfected with control plasmid (expressing only EGFP) and 161±4 bpm when hMSCs were expressing both EGFP+mHCN2 (P<0.05). We next injected 106 hMSCs transfected with either control plasmid or mHCN2 gene construct subepicardially in the canine left ventricular wall in situ. During sinus arrest, all control (EGFP) hearts had spontaneous rhythms (45±1 bpm, 2 of right-sided origin and 2 of left). In the EGFP+mHCN2 group, 5 of 6 animals developed spontaneous rhythms of left-sided origin (rate=61±5 bpm; P<0.05). Moreover, immunostaining of the injected regions demonstrated the presence of hMSCs forming gap junctions with adjacent myocytes. These findings demonstrate that genetically modified hMSCs can express functional HCN2 channels in vitro and in vivo, mimicking overexpression of HCN2 genes in cardiac myocytes, and represent a novel delivery system for pacemaker genes into the heart or other electrical syncytia.
Key Words: gene therapy • heart block • ion channels • pacemakers • stem cells
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