Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity
Promoting cardiomyocyte proliferation is a promising strategy to regenerate the heart. Yet, so far, it is poorly understood how cardiomyocyte proliferation is regulated, and no factor identified to promote mammalian cardiomyocyte proliferation has been translated into medical practice. Therefore, fi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Cardiovascular Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcvm.2022.901396/full |
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| author | Ajit Magadum Ajit Magadum Ajit Magadum Harsha V. Renikunta Harsha V. Renikunta Neha Singh Conchi Estaras Raj Kishore Raj Kishore Felix B. Engel Felix B. Engel Felix B. Engel |
| author_facet | Ajit Magadum Ajit Magadum Ajit Magadum Harsha V. Renikunta Harsha V. Renikunta Neha Singh Conchi Estaras Raj Kishore Raj Kishore Felix B. Engel Felix B. Engel Felix B. Engel |
| author_sort | Ajit Magadum |
| collection | DOAJ |
| description | Promoting cardiomyocyte proliferation is a promising strategy to regenerate the heart. Yet, so far, it is poorly understood how cardiomyocyte proliferation is regulated, and no factor identified to promote mammalian cardiomyocyte proliferation has been translated into medical practice. Therefore, finding a novel factor will be vital. Here, we established a live cell screening based on mouse embryonic stem cell-derived cardiomyocytes expressing a non-functional human geminin deletion mutant fused to Azami Green (CM7/1-hgem-derived cardiomyocytes). We screened for a subset of compounds of the small molecule library Spectrum Collection and identified 19 potential inducers of stem cell-derived cardiomyocyte proliferation. Furthermore, the pro-proliferative potential of identified candidate compounds was validated in neonatal and adult rat cardiomyocytes as well as human induced pluripotent stem cell-derived cardiomyocytes. 18 of these compounds promoted mitosis and cytokinesis in neonatal rat cardiomyocytes. Among the top four candidates were two cardiac glycosides, peruvoside and convallatoxin, the flavonoid osajin, and the selective α-adrenoceptor antagonist and imidazoline I1 receptor ligand efaroxan hydrochloride. Inhibition of PTEN and GSK-3β enhanced cell cycle re-entry and progression upon stimulation with cardiac glycosides and osajin, while inhibition of IP3 receptors inhibited the cell cycle-promoting effect of cardiac glycosides. Collectively, we established a screening system and identified potential compounds to promote cardiomyocyte proliferation. Our data suggest that modulation of calcium handling and metabolism promotes cardiomyocyte proliferation, and cardiac glycosides might, besides increasing myocardial contraction force, contribute to cardiac repair by inducing cardiomyocyte proliferation. |
| first_indexed | 2024-04-12T18:15:37Z |
| format | Article |
| id | doaj.art-1b9e61786d58441c9afa0e7cdbf1797f |
| institution | Directory Open Access Journal |
| issn | 2297-055X |
| language | English |
| last_indexed | 2024-04-12T18:15:37Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cardiovascular Medicine |
| spelling | doaj.art-1b9e61786d58441c9afa0e7cdbf1797f2022-12-22T03:21:39ZengFrontiers Media S.A.Frontiers in Cardiovascular Medicine2297-055X2022-09-01910.3389/fcvm.2022.901396901396Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activityAjit Magadum0Ajit Magadum1Ajit Magadum2Harsha V. Renikunta3Harsha V. Renikunta4Neha Singh5Conchi Estaras6Raj Kishore7Raj Kishore8Felix B. Engel9Felix B. Engel10Felix B. Engel11Department of Cardiac Development and Remodelling, Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, GermanyCardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, NY, United StatesLewis Katz School of Medicine, Center for Translational Medicine, Temple University, Philadelphia, PA, United StatesDepartment of Cardiac Development and Remodelling, Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, GermanyDepartment of Cardiology, Charité Berlin - University Medicine, Berlin, GermanyDepartment of Sports Biosciences, Central University of Rajasthan, Ajmer, IndiaLewis Katz School of Medicine, Center for Translational Medicine, Temple University, Philadelphia, PA, United StatesLewis Katz School of Medicine, Center for Translational Medicine, Temple University, Philadelphia, PA, United StatesDepartment of Cardiovascular Sciences, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, United StatesDepartment of Cardiac Development and Remodelling, Max-Planck-Institute for Heart and Lung Research, Bad Nauheim, GermanyExperimental Renal and Cardiovascular Research, Department of Nephropathology, Institute of Pathology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, GermanyMuscle Research Center Erlangen (MURCE), Erlangen, GermanyPromoting cardiomyocyte proliferation is a promising strategy to regenerate the heart. Yet, so far, it is poorly understood how cardiomyocyte proliferation is regulated, and no factor identified to promote mammalian cardiomyocyte proliferation has been translated into medical practice. Therefore, finding a novel factor will be vital. Here, we established a live cell screening based on mouse embryonic stem cell-derived cardiomyocytes expressing a non-functional human geminin deletion mutant fused to Azami Green (CM7/1-hgem-derived cardiomyocytes). We screened for a subset of compounds of the small molecule library Spectrum Collection and identified 19 potential inducers of stem cell-derived cardiomyocyte proliferation. Furthermore, the pro-proliferative potential of identified candidate compounds was validated in neonatal and adult rat cardiomyocytes as well as human induced pluripotent stem cell-derived cardiomyocytes. 18 of these compounds promoted mitosis and cytokinesis in neonatal rat cardiomyocytes. Among the top four candidates were two cardiac glycosides, peruvoside and convallatoxin, the flavonoid osajin, and the selective α-adrenoceptor antagonist and imidazoline I1 receptor ligand efaroxan hydrochloride. Inhibition of PTEN and GSK-3β enhanced cell cycle re-entry and progression upon stimulation with cardiac glycosides and osajin, while inhibition of IP3 receptors inhibited the cell cycle-promoting effect of cardiac glycosides. Collectively, we established a screening system and identified potential compounds to promote cardiomyocyte proliferation. Our data suggest that modulation of calcium handling and metabolism promotes cardiomyocyte proliferation, and cardiac glycosides might, besides increasing myocardial contraction force, contribute to cardiac repair by inducing cardiomyocyte proliferation.https://www.frontiersin.org/articles/10.3389/fcvm.2022.901396/fullcardiac glycosidescardiomyocyte proliferationcalcium handlinglive cell screening platform AGAzami Greenstem cell |
| spellingShingle | Ajit Magadum Ajit Magadum Ajit Magadum Harsha V. Renikunta Harsha V. Renikunta Neha Singh Conchi Estaras Raj Kishore Raj Kishore Felix B. Engel Felix B. Engel Felix B. Engel Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity Frontiers in Cardiovascular Medicine cardiac glycosides cardiomyocyte proliferation calcium handling live cell screening platform AG Azami Green stem cell |
| title | Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| title_full | Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| title_fullStr | Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| title_full_unstemmed | Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| title_short | Live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| title_sort | live cell screening identifies glycosides as enhancers of cardiomyocyte cell cycle activity |
| topic | cardiac glycosides cardiomyocyte proliferation calcium handling live cell screening platform AG Azami Green stem cell |
| url | https://www.frontiersin.org/articles/10.3389/fcvm.2022.901396/full |
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