NOTCH1 is Critical for Fibroblast-mediated Induction of Cardiomyocyte Specialization into Ventricular Conduction System-like Cells in Vitro

Overview

Journal Sci Rep

Specialty Science

Date 2020 Oct 1

PMID 32999360

Citations 6

Authors

Agatha Ribeiro da Silva

Elida A Neri

Lauro Thiago Turaca

Rafael Dariolli

Miriam H Fonseca-Alaniz

Artur Santos-Miranda

Danilo Roman-Campos

Gabriela Venturini

Jose E Krieger

Affiliations

Soon will be listed here.

Abstract

Cardiac fibroblasts are present throughout the myocardium and are enriched in the microenvironment surrounding the ventricular conduction system (VCS). Several forms of arrhythmias are linked to VCS abnormalities, but it is still unclear whether VCS malformations are cardiomyocyte autonomous or could be linked to crosstalk between different cell types. We reasoned that fibroblasts influence cardiomyocyte specialization in VCS cells. We developed 2D and 3D culture models of neonatal rat cardiac cells to assess the influence of cardiac fibroblasts on cardiomyocytes. Cardiomyocytes adjacent to cardiac fibroblasts showed a two-fold increase in expression of VCS markers (NAV1.5 and CONTACTIN 2) and calcium transient duration, displaying a Purkinje-like profile. Fibroblast-conditioned media (fCM) was sufficient to activate VCS-related genes (Irx3, Scn5a, Connexin 40) and to induce action potential prolongation, a hallmark of Purkinge phenotype. fCM-mediated response seemed to be spatially-dependent as cardiomyocyte organoids treated with fCM had increased expression of connexin 40 and NAV1.5 primarily on its outer surface. Finally, NOTCH1 activation in both cardiomyocytes and fibroblasts was required for connexin 40 up-regulation (a proxy of VCS phenotype). Altogether, we provide evidence that cardiac fibroblasts influence cardiomyocyte specialization into VCS-like cells via NOTCH1 signaling in vitro.

Citing Articles

Analysis of endogenous NOTCH1 from POFUT1 S162L patient fibroblasts reveals the importance of the O-fucose modification on EGF12 in human development.

Matsumoto K, Luther K, Haltiwanger R Glycobiology. 2024; 34(8).

PMID: 38976017 PMC: 11249915. DOI: 10.1093/glycob/cwae047.

Multicellular 3D Models for the Study of Cardiac Fibrosis.

Picchio V, Floris E, Derevyanchuk Y, Cozzolino C, Messina E, Pagano F Int J Mol Sci. 2022; 23(19).

PMID: 36232943 PMC: 9569892. DOI: 10.3390/ijms231911642.

Adriamycin induces cardiac fibrosis in mice via PRMT5-mediated cardiac fibroblast activation.

Dong X, Yuan B, Yu S, Liu H, Pan X, Sun J Acta Pharmacol Sin. 2022; 44(3):573-583.

PMID: 36056082 PMC: 9958096. DOI: 10.1038/s41401-022-00963-x.

The Roles of Cardiac Fibroblasts and Endothelial Cells in Myocarditis.

Xuan Y, Chen C, Wen Z, Wang D Front Cardiovasc Med. 2022; 9:882027.

PMID: 35463742 PMC: 9022788. DOI: 10.3389/fcvm.2022.882027.

Pacemaker translocations and power laws in 2D stem cell-derived cardiomyocyte cultures.

Dunham C, Mackenzie M, Nakano H, Kim A, Juda M, Nakano A PLoS One. 2022; 17(3):e0263976.

PMID: 35286321 PMC: 8920264. DOI: 10.1371/journal.pone.0263976.

References

Pinto A, Ilinykh A, Ivey M, Kuwabara J, DAntoni M, Debuque R . Revisiting Cardiac Cellular Composition. Circ Res. 2015; 118(3):400-9. PMC: 4744092. DOI: 10.1161/CIRCRESAHA.115.307778. View

Ieda M, Tsuchihashi T, Ivey K, Ross R, Hong T, Shaw R . Cardiac fibroblasts regulate myocardial proliferation through beta1 integrin signaling. Dev Cell. 2009; 16(2):233-44. PMC: 2664087. DOI: 10.1016/j.devcel.2008.12.007. View

Hachiro T, Kawahara K, Sato R, Yamauchi Y, Matsuyama D . Changes in the fluctuation of the contraction rhythm of spontaneously beating cardiac myocytes in cultures with and without cardiac fibroblasts. Biosystems. 2007; 90(3):707-15. DOI: 10.1016/j.biosystems.2007.02.009. View

Furtado M, Nim H, Boyd S, Rosenthal N . View from the heart: cardiac fibroblasts in development, scarring and regeneration. Development. 2016; 143(3):387-97. DOI: 10.1242/dev.120576. View

Mohan R, Boukens B, Christoffels V . Developmental Origin of the Cardiac Conduction System: Insight from Lineage Tracing. Pediatr Cardiol. 2018; 39(6):1107-1114. PMC: 6096846. DOI: 10.1007/s00246-018-1906-8. View

Miquerol L, Moreno-Rascon N, Beyer S, Dupays L, Meilhac S, Buckingham M . Biphasic development of the mammalian ventricular conduction system. Circ Res. 2010; 107(1):153-61. DOI: 10.1161/CIRCRESAHA.110.218156. View

Meysen S, Marger L, Hewett K, Jarry-Guichard T, Agarkova I, Chauvin J . Nkx2.5 cell-autonomous gene function is required for the postnatal formation of the peripheral ventricular conduction system. Dev Biol. 2007; 303(2):740-53. DOI: 10.1016/j.ydbio.2006.12.044. View

Cerrone M, Noujaim S, Tolkacheva E, Talkachou A, OConnell R, Berenfeld O . Arrhythmogenic mechanisms in a mouse model of catecholaminergic polymorphic ventricular tachycardia. Circ Res. 2007; 101(10):1039-48. PMC: 2515360. DOI: 10.1161/CIRCRESAHA.107.148064. View

Caref E, Boutjdir M, Himel H, El-Sherif N . Role of subendocardial Purkinje network in triggering torsade de pointes arrhythmia in experimental long QT syndrome. Europace. 2008; 10(10):1218-23. DOI: 10.1093/europace/eun248. View

10.

Kang G, Giovannone S, Liu N, Liu F, Zhang J, Priori S . Purkinje cells from RyR2 mutant mice are highly arrhythmogenic but responsive to targeted therapy. Circ Res. 2010; 107(4):512-9. PMC: 2930621. DOI: 10.1161/CIRCRESAHA.110.221481. View

11.

van Eif V, Devalla H, Boink G, Christoffels V . Transcriptional regulation of the cardiac conduction system. Nat Rev Cardiol. 2018; 15(10):617-630. DOI: 10.1038/s41569-018-0031-y. View

12.

Zhang S, Kim K, Rosen A, Smyth J, Sakuma R, Delgado-Olguin P . Iroquois homeobox gene 3 establishes fast conduction in the cardiac His-Purkinje network. Proc Natl Acad Sci U S A. 2011; 108(33):13576-81. PMC: 3158173. DOI: 10.1073/pnas.1106911108. View

13.

Durocher D, Charron F, Warren R, Schwartz R, Nemer M . The cardiac transcription factors Nkx2-5 and GATA-4 are mutual cofactors. EMBO J. 1997; 16(18):5687-96. PMC: 1170200. DOI: 10.1093/emboj/16.18.5687. View

14.

Linhares V, Almeida N, Menezes D, Elliott D, Lai D, Beyer E . Transcriptional regulation of the murine Connexin40 promoter by cardiac factors Nkx2-5, GATA4 and Tbx5. Cardiovasc Res. 2004; 64(3):402-11. PMC: 3252638. DOI: 10.1016/j.cardiores.2004.09.021. View

15.

Harris B, Spruill L, Edmonson A, Rackley M, Benson D, OBrien T . Differentiation of cardiac Purkinje fibers requires precise spatiotemporal regulation of Nkx2-5 expression. Dev Dyn. 2005; 235(1):38-49. PMC: 2610391. DOI: 10.1002/dvdy.20580. View

16.

Rentschler S, Yen A, Lu J, Petrenko N, Lu M, Manderfield L . Myocardial Notch signaling reprograms cardiomyocytes to a conduction-like phenotype. Circulation. 2012; 126(9):1058-66. PMC: 3607542. DOI: 10.1161/CIRCULATIONAHA.112.103390. View

17.

Milan D, Giokas A, Serluca F, Peterson R, MacRae C . Notch1b and neuregulin are required for specification of central cardiac conduction tissue. Development. 2006; 133(6):1125-32. DOI: 10.1242/dev.02279. View

18.

LaFoya B, Munroe J, Mia M, Detweiler M, Crow J, Wood T . Notch: A multi-functional integrating system of microenvironmental signals. Dev Biol. 2016; 418(2):227-41. PMC: 5144577. DOI: 10.1016/j.ydbio.2016.08.023. View

19.

Abonnenc M, Nabeebaccus A, Mayr U, Barallobre-Barreiro J, Dong X, Cuello F . Extracellular matrix secretion by cardiac fibroblasts: role of microRNA-29b and microRNA-30c. Circ Res. 2013; 113(10):1138-47. DOI: 10.1161/CIRCRESAHA.113.302400. View

20.

Howard C, Baudino T . Dynamic cell-cell and cell-ECM interactions in the heart. J Mol Cell Cardiol. 2013; 70:19-26. DOI: 10.1016/j.yjmcc.2013.10.006. View