A new combination of transcription factors increases the harvesting efficiency of pacemaker‑like cells

Zhang,Jian; Huang,Congxin

doi:10.3892/mmr.2019.10012

A new combination of transcription factors increases the harvesting efficiency of pacemaker‑like cells

Authors:
- Jian Zhang
- Congxin Huang
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Affiliations: Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: March 6, 2019 https://doi.org/10.3892/mmr.2019.10012
Pages: 3584-3592
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Biological pacemakers that combine cell‑based and gene‑based therapies are a promising treatment for sick sinus syndrome or severe atrioventricular block. The current study aimed to induce differentiation of adipose tissue‑derived stem cells (ADSCs) into cardiac pacemaker cells through co‑expression of the transcription factors insulin gene enhancer binding protein 1 (ISL‑1) and T‑box18 (Tbx18). ADSCs were transfected with green fluorescent protein, ISL‑1, Tbx18 or ISL‑1+Tbx18 ﬂuorescent protein lentiviral vectors, and subsequently co‑cultured with neonatal rat ventricular cardiomyocytes in vitro for 7 days. The potential for regulating the differentiation of ADSCs into pacemaker‑like cells was evaluated by cell morphology, beating rate, reverse transcription‑quantitative polymerase chain reaction, western blotting, immunofluorescence and electrophysiological activity. ADSCs were successfully transformed into spontaneously beating cells that exhibited a behavior similar to that of co‑cultured pacemaker cells. This effect was significantly increased in the combined ISL‑1 and Tbx18 group. These results provide a potential strategy for enriching the cardiac pacemaker cell population from ADSCs.

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1	Cho HC and Marbán E: Biological therapies for cardiac arrhythmias: Can genes and cells replace drugs and devices? Circ Res. 106:674–685. 2010. View Article : Google Scholar : PubMed/NCBI
2	Boink GJ, Christoffels VM, Robinson RB and Tan HL: The past, present, and future of pacemaker therapies. Trends Cardiovasc Med. 25:661–673. 2015. View Article : Google Scholar : PubMed/NCBI
3	Cingolani E, Goldhaber JI and Marbán E: Next-generation pacemakers: From small devices to biological pacemakers. Nat Rev Cardiol. 15:139–150. 2018. View Article : Google Scholar : PubMed/NCBI
4	Ionta V, Liang W, Kim EH, Rafie R, Giacomello A, Marbán E and Cho HC: SHOX2 overexpression favors differentiation of embryonic stem cells into cardiac pacemaker cells, improving biological pacing ability. Stem Cell Reports. 4:129–142. 2015. View Article : Google Scholar : PubMed/NCBI
5	Saito Y, Nakamura K, Yoshida M, Sugiyama H, Ohe T, Kurokawa J, Furukawa T, Takano M, Nagase S, Morita H, et al: Enhancement of spontaneous activity by HCN4 overexpression in mouse embryonic stem cell-derived cardiomyocytes-a possible biological pacemaker. PLoS One. 10:e01381932015. View Article : Google Scholar : PubMed/NCBI
6	Yang M, Zhang GG, Wang T, Wang X, Tang YH, Huang H, Barajas-Martinez H, Hu D and Huang CX: TBX18 gene induces adipose-derived stem cells to differentiate into pacemaker-like cells in the myocardial microenvironment. Int J Mol Med. 38:1403–1410. 2016. View Article : Google Scholar : PubMed/NCBI
7	Dmitrieva RI, Minullina IR, Bilibina AA, Tarasova OV, Anisimov SV and Zaritskey AY: Bone marrow- and subcutaneous adipose tissue-derived mesenchymal stem cells: Differences and similarities. Cell Cycle. 11:377–383. 2012. View Article : Google Scholar : PubMed/NCBI
8	Joo HJ, Kim JH and Hong SJ: Adipose tissue-derived stem cells for myocardial regeneration. Korean Circ J. 47:151–159. 2017. View Article : Google Scholar : PubMed/NCBI
9	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
10	Colombo S, de Sena-Tomás C, George V, Werdich AA, Kapur S, MacRae CA and Targoff KL: Nkx genes establish second heart field cardiomyocyte progenitors at the arterial pole and pattern the venous pole through Isl1 repression. Development. 145:2018. View Article : Google Scholar : PubMed/NCBI
11	Blaschke RJ, Hahurij ND, Kuijper S, Just S, Wisse LJ, Deissler K, Maxelon T, Anastassiadis K, Spitzer J, Hardt SE, et al: Targeted mutation reveals essential functions of the homeodomain transcription factor Shox2 in sinoatrial and pacemaking development. Circulation. 115:1830–1838. 2007. View Article : Google Scholar : PubMed/NCBI
12	Christoffels VM, Mommersteeg MT, Trowe MO, Prall OW, de Gier-de Vries C, Soufan AT, Bussen M, Schuster-Gossler K, Harvey RP, Moorman AF, et al: Formation of the venous pole of the heart from an Nkx2-5-negative precursor population requires Tbx18. Circ Res. 98:1555–1563. 2006. View Article : Google Scholar : PubMed/NCBI
13	Mommersteeg MT, Domínguez JN, Wiese C, Norden J, de Gier-de Vries C, Burch JB, Kispert A, Brown NA, Moorman AF and Christoffels VM: The sinus venosus progenitors separate and diversify from the first and second heart fields early in development. Cardiovasc Res. 87:92–101. 2010. View Article : Google Scholar : PubMed/NCBI
14	Lopez MJ and Spencer ND: In vitro adult rat adipose tissue-derived stromal cell isolation and differentiation. Methods Mol Biol. 702:37–46. 2011. View Article : Google Scholar : PubMed/NCBI
15	Lokuta A, Kirby MS, Gaa ST, Lederer WJ and Rogers TB: On establishing primary cultures of neonatal rat ventricular myocytes for analysis over long periods. J Cardiovasc Electrophysiol. 5:50–62. 1994. View Article : Google Scholar : PubMed/NCBI
16	Zhu Y, Liu T, Song K, Ning R, Ma X and Cui Z: ADSCs differentiated into cardiomyocytes in cardiac microenvironment. Mol Cell Biochem. 324:117–129. 2009. View Article : Google Scholar : PubMed/NCBI
17	Choi YS, Dusting GJ, Stubbs S, Arunothayaraj S, Han XL, Collas P, Morrison WA and Dilley RJ: Differentiation of human adipose-derived stem cells into beating cardiomyocytes. J Cell Mol Med. 14:878–889. 2010. View Article : Google Scholar : PubMed/NCBI
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
19	Miake J, Marbán E and Nuss HB: Biological pacemaker created by gene transfer. Nature. 419:132–133. 2002. View Article : Google Scholar : PubMed/NCBI
20	Bucchi A, Plotnikov AN, Shlapakova I, Danilo P Jr, Kryukova Y, Qu J, Lu Z, Liu H, Pan Z, Potapova I, et al: Wild-type and mutant HCN channels in a tandem biological-electronic cardiac pacemaker. Circulation. 114:992–999. 2006. View Article : Google Scholar : PubMed/NCBI
21	Boink GJ, Nearing BD, Shlapakova IN, Duan L, Kryukova Y, Bobkov Y, Tan HL, Cohen IS, Danilo P Jr, Robinson RB, et al: Ca(2+)-stimulated adenylyl cyclase AC1 generates efficient biological pacing as single gene therapy and in combination with HCN2. Circulation. 126:528–536. 2012. View Article : Google Scholar : PubMed/NCBI
22	Ruhparwar A, Tebbenjohanns J, Niehaus M, Mengel M, Irtel T, Kofidis T, Pichlmaier AM and Haverich A: Transplanted fetal cardiomyocytes as cardiac pacemaker. Eur J Cardiothorac Surg. 21:853–857. 2002. View Article : Google Scholar : PubMed/NCBI
23	Kehat I, Khimovich L, Caspi O, Gepstein A, Shofti R, Arbel G, Huber I, Satin J, Itskovitz-Eldor J and Gepstein L: Electromechanical integration of cardiomyocytes derived from human embryonic stem cells. Nat Biotechnol. 22:1282–1289. 2004. View Article : Google Scholar : PubMed/NCBI
24	Xue T, Cho HC, Akar FG, Tsang SY, Jones SP, Marbán E, Tomaselli GF and Li RA: Functional integration of electrically active cardiac derivatives from genetically engineered human embryonic stem cells with quiescent recipient ventricular cardiomyocytes: Insights into the development of cell-based pacemakers. Circulation. 111:11–20. 2005. View Article : Google Scholar : PubMed/NCBI
25	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
26	Liu H, Zhu F, Yong J, Zhang P, Hou P, Li H, Jiang W, Cai J, Liu M, Cui K, et al: Generation of induced pluripotent stem cells from adult rhesus monkey fibroblasts. Cell Stem Cell. 3:587–590. 2008. View Article : Google Scholar : PubMed/NCBI
27	Li W, Wei W, Zhu S, Zhu J, Shi Y, Lin T, Hao E, Hayek A, Deng H and Ding S: Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors. Cell Stem Cell. 4:16–19. 2009. View Article : Google Scholar : PubMed/NCBI
28	Zhang J, Wilson GF, Soerens AG, Koonce CH, Yu J, Palecek SP, Thomson JA and Kamp TJ: Functional cardiomyocytes derived from human induced pluripotent stem cells. Circ Res. 104:e30–e41. 2009. View Article : Google Scholar : PubMed/NCBI