Cardiomyocyte Progenitor Cells as a Functional Gene Delivery Vehicle for Long-Term Biological Pacing
Anna M. D. Végh,
A. Dénise den Haan,
Lucía Cócera Ortega,
Arie O. Verkerk,
Joost P. G. Sluijter,
Diane Bakker,
Shirley van Amersfoorth,
Toon A. B. van Veen,
Mischa Klerk,
Jurgen Seppen,
Jacques M. T. de Bakker,
Vincent M. Christoffels,
Dirk Geerts,
Marie José T. H. Goumans,
Hanno L. Tan,
Gerard J. J. Boink
Affiliations
Anna M. D. Végh
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
A. Dénise den Haan
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Lucía Cócera Ortega
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Arie O. Verkerk
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Joost P. G. Sluijter
Department of Cardiology, Experimental Cardiology Laboratory, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Diane Bakker
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Shirley van Amersfoorth
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Toon A. B. van Veen
Medical Physiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
Mischa Klerk
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Jurgen Seppen
Tytgat Institute, Academic Medical Center, 1105 BK Amsterdam, The Netherlands
Jacques M. T. de Bakker
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Vincent M. Christoffels
Department of Medical Biology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Dirk Geerts
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Marie José T. H. Goumans
Cell and Chemical Biology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands
Hanno L. Tan
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Gerard J. J. Boink
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands
Sustained pacemaker function is a challenge in biological pacemaker engineering. Human cardiomyocyte progenitor cells (CMPCs) have exhibited extended survival in the heart after transplantation. We studied whether lentivirally transduced CMPCs that express the pacemaker current If (encoded by HCN4) can be used as functional gene delivery vehicle in biological pacing. Human CMPCs were isolated from fetal hearts using magnetic beads coated with Sca-1 antibody, cultured in nondifferentiating conditions, and transduced with a green fluorescent protein (GFP)- or HCN4-GFP-expressing lentivirus. A patch-clamp analysis showed a large hyperpolarization-activated, time-dependent inward current (−20 pA/pF at −140 mV, n = 14) with properties typical of If in HCN4-GFP-expressing CMPCs. Gap-junctional coupling between CMPCs and neonatal rat ventricular myocytes (NRVMs) was demonstrated by efficient dye transfer and changes in spontaneous beating activity. In organ explant cultures, the number of preparations showing spontaneous beating activity increased from 6.3% in CMPC/GFP-injected preparations to 68.2% in CMPC/HCN4-GFP-injected preparations (P < 0.05). Furthermore, in CMPC/HCN4-GFP-injected preparations, isoproterenol induced a significant reduction in cycle lengths from 648 ± 169 to 392 ± 71 ms (P < 0.05). In sum, CMPCs expressing HCN4-GFP functionally couple to NRVMs and induce physiologically controlled pacemaker activity and may therefore provide an attractive delivery platform for sustained pacemaker function.