In vitro study of the effects of reprogramming neonatal rat fibroblasts transfected with TBX18 on spontaneous beating in neonatal rat cardiomyocytes

Quan,Dajun; Huang,He

doi:10.3892/mmr.2018.9566

In vitro study of the effects of reprogramming neonatal rat fibroblasts transfected with TBX18 on spontaneous beating in neonatal rat cardiomyocytes

Authors:
- Dajun Quan
- He Huang
View Affiliations

Affiliations: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: October 22, 2018 https://doi.org/10.3892/mmr.2018.9566
Pages: 5520-5526
Copyright: © Quan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Intracardiac injection of the growth‑promoting transcription factor of the sinoatrial node T‑box18 (TBX18) has been shown to reprogram cardiomyocytes into induced sinoatrial nodes that produce high‑frequency and neuroregulated ectopic beats. Fibroblasts, the most important non‑cardiomyocyte cell type in the heart, can affect the electrophysiological properties of cardiomyocytes by electrically coupling with them. The aim of the present study was to explore the reprogramming of cardiac fibroblasts (CFs) transfected with TBX18 in vitro and observe its effect on the pacing frequency of neonatal rat ventricular cardiomyocytes (NRVMs) in a co‑culture system. CFs transfected with TBX18 could be transformed into cardiac myofibroblasts that expressed high levels of hyperpolarization‑activated cyclic nucleotide‑gated cation channel 4 protein and low levels of connexin 43 (COX43) and COX45 protein. In addition, TBX18‑CFs could increase the beating rates of NRVMs and TBX18‑NRVMs in a co‑culture system. The results of the present study indicated that the TBX18 gene could induce CFs to undergo a transformation that promotes an increase of the beating rates of NRVMs and TBX18‑NRVMs.

View Figures

View References

1	Zhou Q and Melton DA: Extreme makeover: Converting one cell into another. Cell Stem Cell. 3:382–388. 2008. View Article : Google Scholar : PubMed/NCBI
2	Vierbuchen T, Ostermeier A, Pang ZP, Kokubu Y, Südhof TC and Wernig M: Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 463:1035–1041. 2010. View Article : Google Scholar : PubMed/NCBI
3	Ieda M, Fu JD, Delgado-Olguin P, Vedantham V, Hayashi Y, Bruneau BG and Srivastava D: Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors. Cell. 142:375–386. 2010. View Article : Google Scholar : PubMed/NCBI
4	Szabo E, Rampalli S, Risueño RM, Schnerch A, Mitchell R, Fiebig-Comyn A, Levadoux-Martin M and Bhatia M: Direct conversion of human fibroblasts to multilineage blood progenitors. Nature. 468:521–526. 2010. View Article : Google Scholar : PubMed/NCBI
5	Hu YF, Dawkins JF, Cho HC, Marbán E and Cingolani E: Biological pacemaker created by minimally invasive somatic reprogramming in pigs with complete heart block. Sci Transl Med. 6:245ra942014. View Article : Google Scholar : PubMed/NCBI
6	Wiese C, Grieskamp T, Airik R, Mommersteeg MT, Gardiwal A, de Gier-de Vries C, Schuster-Gossler K, Moorman AF, Kispert A and Christoffels VM: Formation of the sinus node head and differentiation of sinus node myocardium are independently regulated by Tbx18 and Tbx3. Circ Res. 104:388–397. 2009. View Article : Google Scholar : PubMed/NCBI
7	Kapoor N, Liang W, Marbán E and Cho HC: Direct conversion of quiescent cardiomyocytes to pacemaker cells by expression of Tbx18. Nat Biotechnol. 31:54–62. 2013. View Article : Google Scholar : PubMed/NCBI
8	Ongstad E and Kohl P: Fibroblast-myocyte coupling in the heart: Potential relevance for therapeutic interventions. J Mol Cell Cardiol. 91:238–246. 2016. View Article : Google Scholar : PubMed/NCBI
9	Takahashi K and Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 126:663–676. 2006. View Article : Google Scholar : PubMed/NCBI
10	Weintraub H, Tapscott SJ, Davis RL, Thayer MJ, Adam MA, Lassar AB and Miller AD: Activation of muscle-specific genes in pigment, nerve, fat, liver, and fibroblast cell lines by forced expression of MyoD. Proc Natl Acad Sci USA. 86:5434–5438. 1989. View Article : Google Scholar : PubMed/NCBI
11	Halder G, Callaerts P and Gehring WJ: Induction of ectopic eyes by targeted expression of the eyeless gene in Drosophila. Science. 267:1788–1792. 1995. View Article : Google Scholar : PubMed/NCBI
12	Chambers SM and Studer L: Cell fate plug and play: Direct reprogramming and induced pluripotency. Cell. 145:827–830. 2011. View Article : Google Scholar : PubMed/NCBI
13	Brown TR, Krogh-Madsen T and Christini DJ: Computational approaches to understanding the role of fibroblast-myocyte interactions in cardiac arrhythmogenesis. Biomed Res Int. 2015:4657142015. View Article : Google Scholar : PubMed/NCBI
14	Goldsmith EC, Hoffman A, Morales MO, Potts JD, Price RL, McFadden A, Rice M and Borg TK: Organization of fibroblasts in the heart. Dev Dyn. 230:787–794. 2004. View Article : Google Scholar : PubMed/NCBI
15	Kizana E, Ginn SL, Allen DG, Ross DL and Alexander IE: Fibroblasts can be genetically modified to produce excitable cells capable of electrical coupling. Circulation. 111:394–398. 2005. View Article : Google Scholar : PubMed/NCBI
16	Rosker C, Salvarani N, Schmutz S, Grand T and Rohr S: Abolishing myofibroblast arrhythmogeneicity by pharmacological ablation of α-smooth muscle actin containing stress fibers. Circ Res. 109:1120–1131. 2011. View Article : Google Scholar : PubMed/NCBI
17	Haraguchi Y, Shimizu T, Yamato M and Okano T: Electrical interaction between cardiomyocyte sheets separated by non-cardiomyocyte sheets in heterogeneous tissues. J Tissue Eng Regen Med. 4:291–299. 2010. View Article : Google Scholar : PubMed/NCBI
18	Fahrenbach JP, Mejia-Alvarez R and Banach K: The relevance of non-excitable cells for cardiac pacemaker function. J Physiol. 585:565–578. 2007. View Article : Google Scholar : PubMed/NCBI
19	Abramochkin DV, Lozinsky IT and Kamkin A: Influence of mechanical stress on fibroblast-myocyte interactions in mammalian heart. J Mol Cell Cardiol. 70:27–36. 2014. View Article : Google Scholar : PubMed/NCBI