دورية أكاديمية
Toward Biological Pacing by Cellular Delivery of Hcn2/SkM1.
| العنوان: | Toward Biological Pacing by Cellular Delivery of Hcn2/SkM1. |
|---|---|
| المؤلفون: | Végh AMD; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands.; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Verkerk AO; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.; Department of Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Cócera Ortega L; Department of Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Wang J; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Geerts D; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Klerk M; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Lodder K; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands., Nobel R; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands., Tijsen AJ; Department of Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Devalla HD; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Christoffels VM; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Medina-Ramírez M; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Smits AM; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands., Tan HL; Department of Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.; Netherlands Heart Institute, Utrecht, Netherlands., Wilders R; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands., Goumans MJTH; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands., Boink GJJ; Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands.; Department of Clinical Cardiology, Heart Center, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands. |
| المصدر: | Frontiers in physiology [Front Physiol] 2021 Jan 06; Vol. 11, pp. 588679. Date of Electronic Publication: 2021 Jan 06 (Print Publication: 2020). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Frontiers Research Foundation Country of Publication: Switzerland NLM ID: 101549006 Publication Model: eCollection Cited Medium: Print ISSN: 1664-042X (Print) Linking ISSN: 1664042X NLM ISO Abbreviation: Front Physiol Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Lausanne : Frontiers Research Foundation |
| مستخلص: | Electronic pacemakers still face major shortcomings that are largely intrinsic to their hardware-based design. Radical improvements can potentially be generated by gene or cell therapy-based biological pacemakers. Our previous work identified adenoviral gene transfer of Hcn2 and SkM1, encoding a "funny current" and skeletal fast sodium current, respectively, as a potent combination to induce short-term biological pacing in dogs with atrioventricular block. To achieve long-term biological pacemaker activity, alternative delivery platforms need to be explored and optimized. The aim of the present study was therefore to investigate the functional delivery of Hcn2/SkM1 via human cardiomyocyte progenitor cells (CPCs). Nucleofection of Hcn2 and SkM1 in CPCs was optimized and gene transfer was determined for Hcn2 and SkM1 in vitro . The modified CPCs were analyzed using patch-clamp for validation and characterization of functional transgene expression. In addition, biophysical properties of Hcn2 and SkM1 were further investigated in lentivirally transduced CPCs by patch-clamp analysis. To compare both modification methods in vivo , CPCs were nucleofected or lentivirally transduced with GFP and injected in the left ventricle of male NOD-SCID mice. After 1 week, hearts were collected and analyzed for GFP expression and cell engraftment. Subsequent functional studies were carried out by computational modeling. Both nucleofection and lentiviral transduction of CPCs resulted in functional gene transfer of Hcn2 and SkM1 channels. However, lentiviral transduction was more efficient than nucleofection-mediated gene transfer and the virally transduced cells survived better in vivo . These data support future use of lentiviral transduction over nucleofection, concerning CPC-based cardiac gene delivery. Detailed patch-clamp studies revealed Hcn2 and Skm1 current kinetics within the range of previously reported values of other cell systems. Finally, computational modeling indicated that CPC-mediated delivery of Hcn2/SkM1 can generate stable pacemaker function in human ventricular myocytes. These modeling studies further illustrated that SkM1 plays an essential role in the final stage of diastolic depolarization, thereby enhancing biological pacemaker functioning delivered by Hcn2. Altogether these studies support further development of CPC-mediated delivery of Hcn2/SkM1 and functional testing in bradycardia models. Competing Interests: HT and GB report ownership interest in PacingCure B.V. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 Végh, Verkerk, Cócera Ortega, Wang, Geerts, Klerk, Lodder, Nobel, Tijsen, Devalla, Christoffels, Medina-Ramírez, Smits, Tan, Wilders, Goumans and Boink.) |
| References: | Circulation. 2002 Jan 1;105(1):93-8. (PMID: 11772882) Circ Arrhythm Electrophysiol. 2017 May;10(5):e004508. (PMID: 28500172) Cardiovasc Res. 2004 Oct 1;64(1):12-23. (PMID: 15364609) Mol Ther. 2004 Oct;10(4):768-79. (PMID: 15451461) Heart Rhythm. 2010 Dec;7(12):1835-40. (PMID: 20708103) Pacing Clin Electrophysiol. 2013 Sep;36(9):1138-49. (PMID: 23663261) Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11498-502. (PMID: 8265580) J Am Coll Cardiol. 2011 Aug 30;58(10):995-1000. (PMID: 21867832) Heart Rhythm. 2008 Feb;5(2):282-8. (PMID: 18242555) Nat Biotechnol. 2004 Oct;22(10):1282-9. (PMID: 15448703) Am J Physiol Heart Circ Physiol. 2006 Sep;291(3):H1088-100. (PMID: 16565318) Circ Cardiovasc Imaging. 2008 Sep;1(2):94-103. (PMID: 19808526) Mol Imaging Biol. 2010 Jan-Feb;12(1):15-24. (PMID: 19551446) Circulation. 2005 Jan 4;111(1):11-20. (PMID: 15611367) Circulation. 2006 Sep 5;114(10):992-9. (PMID: 16923750) Mol Biotechnol. 2009 Nov;43(3):250-6. (PMID: 19598009) Cytotherapy. 2012 May;14(5):529-39. (PMID: 22316056) Stem Cells Dev. 2006 Feb;15(1):87-96. (PMID: 16522166) Circ Arrhythm Electrophysiol. 2012 Aug 1;5(4):831-40. (PMID: 22722661) J Biol Chem. 2004 Oct 15;279(42):43497-502. (PMID: 15292247) J Physiol. 2013 Sep 1;591(17):4189-206. (PMID: 23878377) J Am Coll Cardiol. 2013 Mar 19;61(11):1192-201. (PMID: 23395072) Nat Biotechnol. 2017 Jan;35(1):56-68. (PMID: 27941801) Circulation. 2012 Jul 31;126(5):528-36. (PMID: 22753192) Cardiovasc Res. 2009 Aug 1;83(3):527-35. (PMID: 19429921) Am J Physiol Heart Circ Physiol. 2004 Apr;286(4):H1573-89. (PMID: 14656705) Nat Protoc. 2009;4(2):232-43. (PMID: 19197267) Ann Transl Med. 2015 Mar;3(3):42. (PMID: 25815303) Molecules. 2019 Jan 05;24(1):. (PMID: 30621310) Hum Gene Ther. 2013 Oct;24(10):840-51. (PMID: 23987185) J Membr Biol. 1991 Apr;121(2):101-17. (PMID: 1715403) Front Cardiovasc Med. 2019 Jun 19;6:81. (PMID: 31275946) Stem Cell Res Ther. 2016 Apr 30;7(1):67. (PMID: 27137910) Nat Med. 1996 May;2(5):545-50. (PMID: 8616713) Circulation. 2007 Aug 14;116(7):706-13. (PMID: 17646577) Circ Res. 2004 Apr 16;94(7):952-9. (PMID: 14988226) Circ Res. 2016 Apr 15;118(8):1223-32. (PMID: 26888636) J Physiol. 2009 Nov 1;587(Pt 21):5211-26. (PMID: 19736302) |
| فهرسة مساهمة: | Keywords: HCN channels; SkM1 channels; biological pacemaker; cell therapy; gene therapy; progenitor cells |
| تواريخ الأحداث: | Date Created: 20210125 Latest Revision: 20210126 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC7815531 |
| DOI: | 10.3389/fphys.2020.588679 |
| PMID: | 33488393 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1664-042X |
|---|---|
| DOI: | 10.3389/fphys.2020.588679 |