The epicardium obscures interpretations on endothelial-to-mesenchymal transition in the mouse atrioventricular canal explant assay
Open accessAbstract
Atrioventricular septal defects often result from impaired endocardial cushion development. Endothelial-to-mesenchymal transition (EndoMT) is a critical event in endocardial cushion development that initiates in the atrioventricular canal (AVC). In ex vivo EndoMT studies, mouse AVCs are flat-mounted on a collagen gel. In the explant outgrowths, the ratio of elongated spindle-like mesenchymal cells over cobblestone-shaped cells, generally considered as endothelial cells, reflects EndoMT. Using this method, several key signalling pathways have been attributed important functions during EndoMT. Using genetic lineage tracing and cell-type-specific markers, we show that monolayers of cobblestone-shaped cells are predominantly of epicardial rather than endothelial origin. Furthermore, this epicardium is competent to undergo mesenchymal transition. Contamination by epicardium is common and inherent as this tissue progressively attaches to AVC myocardium. Inhibition of TGFβ signalling, previously shown to blunt EndoMT, caused an enrichment in epicardial monolayers. The presence of epicardium thus confounds interpretations of EndoMT signalling pathways in this assay. We advocate to systematically use lineage tracers and cell-type-specific markers on stage-matched AVC explants. Furthermore, a careful reconsideration of earlier studies on EndoMT using this explant assay may identify unanticipated epicardial effects and/or the presence of epicardial-to-mesenchymal transition (EpiMT), which would alter the interpretation of results on endothelial-to-mesenchymal transition.
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The presence of endothelial and epicardial lineages in the AVC confounds interpretations of cell signalling on each process individually.
“especially as the presence of endothelial and epicardial lineages in the AVC confounds interpretations of cell signalling on each of these processes individually65.”
From Cell diversity and plasticity during atrioventricular heart valve EMTs (2023) · cites this paper as [65]
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Cites
Tie2-Cre Transgenic Mice: A New Model for Endothelial Cell-Lineage Analysis in Vivo
Kisanuki, Y. Y., Hammer, R. E., Miyazaki, J.-I., Williams, S. C., Richardson, J. A., & Yanagisawa, M. (2001) · Developmental Biology · doi:10.1006/dbio.2000.0106 · Cited by 1,241 · see citing works
Ma, L., Lu, M.-F., Schwartz, R. J., & Martin, J. F. (2005) · Development · doi:10.1242/dev.02156 · Cited by 525 · see citing works
Uhl, M., Aulwurm, S., Wischhusen, J., Weiler, M., Jing, Y., Almirez, R. G., Mangadu, R., Liu, Y.-W., Platten, M., Herrlinger, U., et al. (2004) · Cancer Research · doi:10.1158/0008-5472.can-04-1013 · Cited by 414 · see citing works
Slug is a direct Notch target required for initiation of cardiac cushion cellularization
Niessen, K., Fu, Y., Chang, L., Hoodless, P. A., McFadden, D., & Karsan, A. (2008) · The Journal of Cell Biology · doi:10.1083/jcb.200710067 · Cited by 363 · see citing works
Camenisch, T. D., Schroeder, J. A., Bradley, J. M., Klewer, S. E., & McDonald, J. A. (2002) · Nature Medicine · doi:10.1038/nm742 · Cited by 330 · see citing works
Characterization of Nkx6-2-Derived Neocortical Interneuron Lineages
Sousa, V. H., Miyoshi, G., Hjerling-Leffler, J., Karayannis, T., & Fishell, G. (2009) · Cerebral Cortex · doi:10.1093/cercor/bhp038 · Cited by 328 · see citing works
Model systems for the study of heart development and disease
Hutson, M. R., & Kirby, M. L. (2006) · Seminars in Cell and Developmental Biology · doi:10.1016/j.semcdb.2006.12.004 · Cited by 306 · see citing works
Camenisch, T. D., Molin, D. G. M., Person, A., Runyan, R. B., Gittenberger-de Groot, A. C., McDonald, J. A., & Klewer, S. E. (2002) · Developmental Biology · doi:10.1006/dbio.2002.0731 · Cited by 279 · see citing works
Luna‐Zurita, L., Prados, B., Grego‐Bessa, J., Luxán, G., del Monte‐Nieto, G., Benguría, A., Adams, R. H., Pérez-Pomares, J. M., & de la Pompa, J. L. (2010) · Journal of Clinical Investigation · doi:10.1172/jci42666 · Cited by 242 · see citing works
Dor, Y., Camenisch, T. D., Itin, A., Fishman, G. I., McDonald, J. A., Carmeliet, P., & Keshet, E. (2001) · Development · doi:10.1242/dev.128.9.1531 · Cited by 240 · see citing works
Sugi, Y., Yamamura, H., Okagawa, H., & Markwald, R. R. (2004) · Developmental Biology · doi:10.1016/j.ydbio.2004.01.045 · Cited by 185 · see citing works
Nfatc1 Coordinates Valve Endocardial Cell Lineage Development Required for Heart Valve Formation
Wu, B., Wang, Y., Lui, W., Langworthy, M., Tompkins, K. L., Hatzopoulos, A. K., Baldwin, H. S., & Zhou, B. (2011) · Circulation Research · doi:10.1161/circresaha.111.245035 · Cited by 182 · see citing works
Migratory behavior of cardiac cushion tissue cells in a collagen-lattice culture system
Bernanke, D. H., & Markwald, R. R. (1982) · Developmental Biology · doi:10.1016/0012-1606(82)90030-6 · Cited by 182 · see citing works
Co-ordinating Notch, BMP, and TGF-β signaling during heart valve development
Garside, V. C., Chang, A. C., Karsan, A., & Hoodless, P. A. (2013) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-012-1197-9 · Cited by 160 · see citing works
Signaling via the Tgf-β type I receptor Alk5 in heart development
Sridurongrit, S., Larsson, J., Schwartz, R. J., Ruiz‐Lozano, P., & Kaartinen, V. (2008) · Developmental Biology · doi:10.1016/j.ydbio.2008.07.038 · Cited by 158 · see citing works
Atrioventricular cushion transformation is mediated by ALK2 in the developing mouse heart
Wang, J., Sridurongrit, S., Dudáš, M., Thomas, P. S., Nagy, A. R., Schneider, M. D., Epstein, J. A., & Kaartinen, V. (2005) · Developmental Biology · doi:10.1016/j.ydbio.2005.07.035 · Cited by 158 · see citing works
Guadix, J. A., Ruiz‐Villalba, A., Lettice, L. A., Velecela, V., Muñoz‐Chápuli, R., Hastie, N. D., Pérez-Pomares, J. M., & Martínez-Estrada, O. M. (2011) · Development · doi:10.1242/dev.044594 · Cited by 134 · see citing works
A heart segmental defect in the anterior-posterior axis of a transgenic mutant mouse
Yamamura, H., Zhang, M., Markwald, R. R., & Mjaatvedt, C. H. (1997) · Developmental Biology · doi:10.1006/dbio.1997.8559 · Cited by 131 · see citing works
MacGrogan, D., D’Amato, G., Travisano, S. I., Martı́nez-Poveda, B., Luxán, G., del Monte‐Nieto, G., Papoutsi, T., Sbroggió, M., Bou, V., Arco, P. G., et al. (2016) · Circulation Research · doi:10.1161/circresaha.115.308077 · Cited by 124 · see citing works
Azhar, M., Runyan, R. B., Gard, C., Sanford, L. P., Miller, M. L., Andringa, A., Pawlowski, S. A., Rajan, S., & Doetschman, T. (2009) · Developmental Dynamics · doi:10.1002/dvdy.21854 · Cited by 123 · see citing works
Kruithof, B. P. T., Krawitz, S. A., & Gaussin, V. (2006) · Developmental Biology · doi:10.1016/j.ydbio.2006.09.024 · Cited by 113 · see citing works
Hofmann, J. J., Briot, A., Enciso, J., Zovein, A. C., Ren, S. V., Zhang, Z. W., Radtke, F., Simons, M., Wang, Y., & Iruela‐Arispe, M. L. (2012) · Development · doi:10.1242/dev.084871 · Cited by 110 · see citing works
Cardiac endothelial cells express Wilms' tumor-1
Duim, S. N., Kurakula, K., Goumans, M., & Kruithof, B. P. T. (2015) · Journal of Molecular and Cellular Cardiology · doi:10.1016/j.yjmcc.2015.02.007 · Cited by 100 · see citing works
Elevated glucose inhibits VEGF-A–mediated endocardial cushion formation
Enciso, J., Gratzinger, D. A., Camenisch, T. D., Canosa, S., Pinter, E., & Madri, J. A. (2003) · The Journal of Cell Biology · doi:10.1083/jcb.200209014 · Cited by 95 · see citing works
Ishii, Y., Garriock, R. J., Navetta, A. M., Coughlin, L. E., & Mikawa, T. (2010) · Developmental Cell · doi:10.1016/j.devcel.2010.07.017 · Cited by 83 · see citing works
Endoglin and Alk5 regulate epithelial–mesenchymal transformation during cardiac valve formation
Mercado-Pimentel, M. E., Hubbard, A. D., & Runyan, R. B. (2006) · Developmental Biology · doi:10.1016/j.ydbio.2006.12.038 · Cited by 75 · see citing works
Effects of hyaluronic acid on cardiac cushion tissue cells in collagen matrix cultures.
Bernanke, D. H., & Markwald, R. R. (1979) · PubMed · Cited by 72 · see citing works
VEGF modulates early heart valve formation
Dor, Y., Klewer, S. E., McDonald, J. A., Keshet, E., & Camenisch, T. D. (2003) · The Anatomical Record Part A Discoveries in Molecular Cellular and Evolutionary Biology · doi:10.1002/ar.a.10026 · Cited by 66 · see citing works
Akt promotes Endocardial-Mesenchyme Transition
Meadows, K. N., Iyer, S., Stevens, M. V., Wang, D., Shechter, S., Perruzzi, C. A., Camenisch, T. D., & Benjamin, L. E. (2009) · Vascular Cell · doi:10.1186/2040-2384-1-2 · Cited by 43 · see citing works
Vicente‐Steijn, R., Scherptong, R. W. C., Kruithof, B. P. T., Duim, S. N., Goumans, M., Wisse, L. J., Zhou, B., Pu, W. T., Poelmann, R. E., Schalij, M. J., et al. (2015) · PLoS ONE · doi:10.1371/journal.pone.0136025 · Cited by 29 · see citing works
Beets, K., Staring, M. W., Criem, N., Maas, E., Schellinx, N., Lopes, S. M. C. D. S., Umans, L., & Zwijsen, A. (2016) · BMC Developmental Biology · doi:10.1186/s12861-016-0133-x · Cited by 24 · see citing works
Self-citationIsolation and culture of mouse proepicardium using serum-free conditions
Garriock, R. J., Mikawa, T., & Yamaguchi, T. P. (2013) · Methods · doi:10.1016/j.ymeth.2013.06.030 · Cited by 13 · see citing works
In vitro model for mouse coronary vasculogenesis
Watanabe, N., Nakagawa, M., Hanato, T., Takeuchi, Y., Hara, M., Yoshida, T., & Imanaka‐Yoshida, K. (2006) · The Anatomical Record Part A Discoveries in Molecular Cellular and Evolutionary Biology · doi:10.1002/ar.a.20340 · Cited by 3 · see citing works
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Gunawan, F., Gentile, A., Fukuda, R., Tsedeke, A. T., Jiménez-Amilburu, V., Ramadass, R., Iida, A., Sehara‐Fujisawa, A., & Stainier, D. Y. R. (2019) · The Journal of Cell Biology · doi:10.1083/jcb.201807175 · Cited by 69 · see citing works
Cell diversity and plasticity during atrioventricular heart valve EMTs
Lotto, J., Cullum, R., Drissler, S., Arostegui, M., Garside, V. C., Fuglerud, B. M., Clement-Ranney, M., Thakur, A., Underhill, T. M., & Hoodless, P. A. (2023) · Nature Communications · doi:10.1038/s41467-023-41279-6 · Cited by 13 · see citing works
“Identifying factors speci fi c in driving EndMT and EpiMT initiation and progression is of interest, especiallyas the presenceof endothelial and epicardial lineages in the AVC confounds interpretations of cell signalling on each of these processes individually 65 .”
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Li, J., He, Y., Bu, H., Wang, M., Yu, J., Li, L., Li, H., Zhang, X., Cui, X., & Cheng, M. (2020) · Heart and Vessels · doi:10.1007/s00380-020-01625-w · Cited by 9 · see citing works
- Authors
- Criem, N., & Zwijsen, A.
- Affiliations
- VIB-KU Leuven Center for Brain & Disease Research (BE)
- Corresponding author
- Zwijsen, An
- Venue
- Scientific Reports
- Published
- 2018-03-16
- Volume
- 8
- Issue
- 1
- Pages
- 4722-4722
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC5856756
- OpenAlex
- W2803404075
- Cited by
- 3
- References
- 33
- Citations by year
- 2023: 1 · 2020: 1 · 2019: 1
- FWCI
- 0.18
- Type
- journal article
- Language
- en
- Update notices
- correction (2020-03-31)
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Lirias (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- Fonds Wetenschappelijk Onderzoek (G078614N)
- KU Leuven (C12/16/023)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Congenital heart defects research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac electrophysiology and arrhythmias
- Keywords
- endothelial-to-mesenchymal transition, epicardium, atrioventricular canal, atrioventricular septal defect, TGF-beta signaling, lineage tracing, cell type-specific markers, mouse heart development
- MeSH terms
- Animals, Atrioventricular Node, Biological Assay, Embryo, Mammalian, Endothelium, Vascular, Female, Male, Pericardium, Signal Transduction, Mice, Rats, Epithelial-Mesenchymal Transition
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