Nkx2.5 insufficiency leads to atrial electrical remodeling through Wnt signaling in HL‑1 cells

Chen,Jingjing; Xu,Shunen; Li,Wei; Wu,Lirong; Wang,Long; Li,Yongkang; Zhou,Wei

doi:10.3892/etm.2019.8134

Nkx2.5 insufficiency leads to atrial electrical remodeling through Wnt signaling in HL‑1 cells

Authors:
- Jingjing Chen
- Shunen Xu
- Wei Li
- Lirong Wu
- Long Wang
- Yongkang Li
- Wei Zhou
View Affiliations

Affiliations: Department of Cardiology, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China, Department of Orthopedics, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China
Published online on: October 25, 2019 https://doi.org/10.3892/etm.2019.8134
Pages: 4631-4636
Copyright: © Chen 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

Homeobox protein Nxk‑2.5 (Nkx2.5) is a homeobox transcription factor that promotes chamber‑like myocardial gene expression. Data from a previous genome‑wide association study suggested that Nkx2.5 may be associated with the genetic variation that underlies atrial fibrillation (AF). Nkx2.5 loss of function has been demonstrated to be associated with an increasing susceptibility of familial AF. Therefore, the aim of the present study was to investigate the effect of Nkx2.5 loss of function on electrophysiological substrates in HL‑1 cells. To the best of our knowledge, the results demonstrated for the first time that Nkx2.5 expression was significantly decreased in a rat model exhibiting AF. The effect of silencing Nkx2.5 was assessed following transfection with adenoviral vectors with specific NKX2.5‑shRNA. The effect of Nkx2.5 silencing on potassium/sodium hyperpolarization‑activated cyclic nucleotide‑gated channel 4 (HCN4), gap junction alpha‑5 protein (Cx40), calcium handling proteins and protein Wnt‑11 (Wnt11) expression levels was also assessed in HL‑1 cells. The results revealed that Nkx2.5 silencing increased HCN4 expression, decreased Cx40 expression and disrupted the expression of calcium handling proteins. Additionally, Wnt11 signal protein expression was decreased following Nkx2.5 silencing. The results of the present study demonstrated that Nkx2.5 served as a transcriptional regulator of the electrophysiological substrates associated with AF.

View Figures

View References

1	Wilke T, Groth A, Mueller S, Pfannkuche M, Verheyen F, Linder R, Maywald U, Bauersachs R and Breithardt G: Incidence and prevalence of atrial fibrillation: An analysis based on 8.3 million patients. Europace. 15:486–493. 2013. View Article : Google Scholar : PubMed/NCBI
2	Andrade J, Khairy P, Dobrev D and Nattel S: The clinical profile and pathophysiology of atrial fibrillation: Relationships among clinical features, epidemiology, and mechanisms. Circ Res. 114:1453–1468. 2014. View Article : Google Scholar : PubMed/NCBI
3	Ye W, Wang J, Song Y, Yu D, Sun C, Liu C, Chen F, Zhang Y, Wang F, Harvey RP, et al: A common Shox2-Nkx2-5 antagonistic mechanism primes the pacemaker cell fate in the pulmonary vein myocardium and sinoatrial node. Development. 142:2521–2532. 2015. View Article : Google Scholar : PubMed/NCBI
4	Nielsen JB, Thorolfsdottir RB, Fritsche LG, Zhou W, Skov MW, Graham SE, Herron TJ, McCarthy S, Schmidt EM, Sveinbjornsson G, et al: Biobank-driven genomic discovery yields new insight into atrial fibrillation biology. Nat Genet. 50:1234–1239. 2018. View Article : Google Scholar : PubMed/NCBI
5	Huang RT, Xue S, Xu YJ, Zhou M and Yang YQ: A novel NKX2.5 loss-of-function mutation responsible for familial atrial fibrillation. Int J Mol Med. 31:1119–1126. 2013. View Article : Google Scholar : PubMed/NCBI
6	National Institutes of Health, . Guide for the Care and Use of Laboratory Animals. 8th. National Academies Press; Washington: pp. 86–23. 2011
7	Claycomb WC, Lanson NA Jr, Stallworth BS, Egeland DB, Delcarpio JB, Bahinski A and Izzo NJ Jr: HL-1 cells: A cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. Proc Natl Acad Sci USA. 95:2979–2984. 1998. View Article : Google Scholar : PubMed/NCBI
8	Lozano-Velasco E, Hernandez-Torres F, Daimi H, Serra SA, Herraiz A, Hove-Madsen L, Aránega A and Franco D: Pitx2 impairs calcium handling in a dose-dependent manner by modulating Wnt signalling. Cardiovasc Res. 109:55–66. 2016. View Article : Google Scholar : PubMed/NCBI
9	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
10	Cardin S, Guasch E, Luo X, Naud P, Le Quang K, Shi Y, Tardif JC, Comtois P and Nattel S: Role for MicroRNA-21 in atrial profibrillatory fibrotic remodeling associated with experimental postinfarction heart failure. Circ Arrhythm Electrophysiol. 5:1027–1035. 2012. View Article : Google Scholar : PubMed/NCBI
11	DiFrancesco D: HCN4, sinus bradycardia and atrial fibrillation. Arrhythm Electrophysiol Rev. 4:9–13. 2015. View Article : Google Scholar : PubMed/NCBI
12	Li YD, Hong YF, Zhang Y, Zhou XH, Ji YT, Li HL, Hu GJ, Li JX, Sun L, Zhang JH, et al: Association between reversal in the expression of hyperpolarization-activated cyclic nucleotide-gated (HCN) channel and age-related atrial fibrillation. Med Sci Monit. 20:2292–2297. 2014. View Article : Google Scholar : PubMed/NCBI
13	Yang YQ, Liu X, Zhang XL, Wang XH, Tan HW, Shi HF, Jiang WF and Fang WY: Novel connexin40 missense mutations in patients with familial atrial fibrillation. Europace. 12:1421–1427. 2010. View Article : Google Scholar : PubMed/NCBI
14	Sun Y, Yang YQ, Gong XQ, Wang XH, Li RG, Tan HW, Liu X, Fang WY and Bai D: Novel germline GJA5/connexin40 mutations associated with lone atrial fibrillation impair gap junctional intercellular communication. Hum Mutat. 34:603–609. 2013.PubMed/NCBI
15	Lübkemeier I, Andrié R, Lickfett L, Bosen F, Stöckigt F, Dobrowolski R, Draffehn AM, Fregeac J, Schultze JL, Bukauskas FF, et al: The Connexin40A96S mutation from a patient with atrial fibrillation causes decreased atrial conduction velocities and sustained episodes of induced atrial fibrillation in mice. J Mol Cell Cardiol. 65:19–32. 2013. View Article : Google Scholar : PubMed/NCBI
16	Houser SR: Role of RyR2 phosphorylation in heart failure and arrhythmias: Protein kinase A-mediated hyperphosphorylation of the ryanodine receptor at serine 2808 does not alter cardiac contractility or cause heart failure and arrhythmias. Circ Res. 114:1320–1327; discussion 1327. 2014. View Article : Google Scholar : PubMed/NCBI
17	Samuel TJ, Rosenberry RP, Lee S and Pan Z: Correcting calcium dysregulation in chronic heart failure using SERCA2a gene therapy. Int J Mol Sci. 19:2018. View Article : Google Scholar
18	Bisson JA, Mills B, Paul Helt JC, Zwaka TP and Cohen ED: Wnt5a and Wnt11 inhibit the canonical Wnt pathway and promote cardiac progenitor development via the Caspase-dependent degradation of AKT. Dev Biol. 398:80–96. 2015. View Article : Google Scholar : PubMed/NCBI
19	Cohen ED, Miller MF, Wang Z, Moon RT and Morrisey EE: Wnt5a and Wnt11 are essential for second heart field progenitor development. Development. 139:1931–1940. 2012. View Article : Google Scholar : PubMed/NCBI
20	Terami H, Hidaka K, Katsumata T, Iio A and Morisaki T: Wnt11 facilitates embryonic stem cell differentiation to Nkx2.5-positive cardiomyocytes. Biochem Biophys Res Commun. 325:968–975. 2004. View Article : Google Scholar : PubMed/NCBI
21	Nagy II, Railo A, Rapila R, Hast T, Sormunen R, Tavi P, Räsänen J and Vainio SJ: Wnt-11 signalling controls ventricular myocardium development by patterning N-cadherin and beta-catenin expression. Cardiovasc Res. 85:100–109. 2010. View Article : Google Scholar : PubMed/NCBI
22	Panáková D, Werdich AA and Macrae CA: Wnt11 patterns a myocardial electrical gradient through regulation of the L-type Ca(2+) channel. Nature. 466:874–878. 2010. View Article : Google Scholar : PubMed/NCBI