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Molecules. 2019 Jan 5;24(1). pii: E181. doi: 10.3390/molecules24010181.

Cardiomyocyte Progenitor Cells as a Functional Gene Delivery Vehicle for Long-Term Biological Pacing.

Author information

1
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. a.m.d.vegh@lumc.nl.
2
Cell and Chemical Biology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands. a.m.d.vegh@lumc.nl.
3
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. denisedenhaan@gmail.com.
4
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. l.coceraortega@amc.uva.nl.
5
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. a.o.verkerk@amc.uva.nl.
6
Department of Cardiology, Experimental Cardiology Laboratory, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands. jsluijter@umcutrecht.nl.
7
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. diane.bakker@amc.uva.nl.
8
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. s.c.vanamersfoorth@amc.uva.nl.
9
Medical Physiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands. s.c.vanamersfoorth@amc.uva.nl.
10
Medical Physiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands. a.a.b.vanveen@umcutrecht.nl.
11
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. m.klerk@amc.uva.nl.
12
Tytgat Institute, Academic Medical Center, 1105 BK Amsterdam, The Netherlands. j.seppen@amc.uva.nl.
13
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. j.m.debakker@amc.uva.nl.
14
Medical Physiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands. j.m.debakker@amc.uva.nl.
15
Department of Medical Biology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. v.m.christoffels@amc.uva.nl.
16
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. h.a.geerts@amc.uva.nl.
17
Department of Medical Biology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. h.a.geerts@amc.uva.nl.
18
Cell and Chemical Biology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands. M.J.T.H.Goumans@lumc.nl.
19
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. h.l.tan@amc.uva.nl.
20
Heart Center, Clinical & Experimental Cardiology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. g.j.boink@amc.uva.nl.
21
Department of Medical Biology, Amsterdam University Medical Centers, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands. g.j.boink@amc.uva.nl.

Abstract

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.

KEYWORDS:

HCN channels; cell therapy; gene therapy; pacemakers; progenitor cells

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