TY - JOUR
T1 - Cardiomyocyte Progenitor Cells as a Functional Gene Delivery Vehicle for Long-Term Biological Pacing
AU - Végh, Anna M D
AU - den Haan, A Dénise
AU - Cócera Ortega, Lucía
AU - Verkerk, Arie O
AU - Sluijter, Joost P G
AU - Bakker, Diane
AU - van Amersfoorth, Shirley
AU - van Veen, Toon A B
AU - Klerk, Mischa
AU - Seppen, Jurgen
AU - de Bakker, Jacques M T
AU - Christoffels, Vincent M
AU - Geerts, Dirk
AU - Goumans, Marie José T H
AU - Tan, Hanno L
AU - Boink, Gerard J J
N1 - Funding Information: Funding: This work was supported by the Rembrandt Institute for Cardiovascular Research [to G.J.J.B., M.J.G., and H.L.T.]. G.J.J.B. is supported by personal grants from the Hartstichting (2014T065), the Nederlandse Organisatie voor Wetenschappelijk Onderzoek (ZonMw Veni 016.156.162), and the European Research Council (ERC Starting Grant 714866). Publisher Copyright: © 2019 by the authors. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2019/1/5
Y1 - 2019/1/5
N2 - 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.
AB - 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.
KW - Animals
KW - Cell therapy
KW - Gene Transfer Techniques
KW - Gene therapy
KW - Genetic Therapy/methods
KW - Green Fluorescent Proteins/chemistry
KW - HCN channels
KW - Heart Ventricles/pathology
KW - Humans
KW - Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/genetics
KW - Muscle Proteins/genetics
KW - Myocytes, Cardiac/transplantation
KW - Pacemakers
KW - Patch-Clamp Techniques
KW - Potassium Channels/genetics
KW - Progenitor cells
KW - Rats
KW - Stem Cell Transplantation
KW - Stem Cells/cytology
UR - http://www.scopus.com/inward/record.url?scp=85059795303&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/molecules24010181
DO - https://doi.org/10.3390/molecules24010181
M3 - Article
C2 - 30621310
SN - 1420-3049
VL - 24
JO - Molecules
JF - Molecules
IS - 1
M1 - 181
ER -