Loss of Ccbe1 Affects Cardiac-specification and Cardiomyocyte Differentiation in Mouse Embryonic Stem Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2018 Oct 4

PMID 30281646

Citations 4

Authors

Oriol Bover

Tiago Justo

Paulo N G Pereira

Joao Facucho-Oliveira

Jose M Inacio

Jose S Ramalho

Ibrahim J Domian

Jose Antonio Belo

Affiliations

Soon will be listed here.

Abstract

Understanding the molecular pathways regulating cardiogenesis is crucial for the early diagnosis of heart diseases and improvement of cardiovascular disease. During normal mammalian cardiac development, collagen and calcium-binding EGF domain-1 (Ccbe1) is expressed in the first and second heart field progenitors as well as in the proepicardium, but its role in early cardiac commitment remains unknown. Here we demonstrate that during mouse embryonic stem cell (ESC) differentiation Ccbe1 is upregulated upon emergence of Isl1- and Nkx2.5- positive cardiac progenitors. Ccbe1 is markedly enriched in Isl1-positive cardiac progenitors isolated from ESCs differentiating in vitro or embryonic hearts developing in vivo. Disruption of Ccbe1 activity by shRNA knockdown or blockade with a neutralizing antibody results in impaired differentiation of embryonic stem cells along the cardiac mesoderm lineage resulting in a decreased expression of mature cardiomyocyte markers. In addition, knockdown of Ccbe1 leads to smaller embryoid bodies. Collectively, our results show that CCBE1 is essential for the commitment of cardiac mesoderm and consequently, for the formation of cardiac myocytes in differentiating mouse ESCs.

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References

Hagerling R, Pollmann C, Andreas M, Schmidt C, Nurmi H, Adams R . A novel multistep mechanism for initial lymphangiogenesis in mouse embryos based on ultramicroscopy. EMBO J. 2013; 32(5):629-44. PMC: 3590982. DOI: 10.1038/emboj.2012.340. View

Jakus Z, Gleghorn J, Enis D, Sen A, Chia S, Liu X . Lymphatic function is required prenatally for lung inflation at birth. J Exp Med. 2014; 211(5):815-26. PMC: 4010903. DOI: 10.1084/jem.20132308. View

Roukens M, Peterson-Maduro J, Padberg Y, Jeltsch M, Leppanen V, Bos F . Functional Dissection of the CCBE1 Protein: A Crucial Requirement for the Collagen Repeat Domain. Circ Res. 2015; 116(10):1660-9. DOI: 10.1161/CIRCRESAHA.116.304949. View

Laugwitz K, Moretti A, Caron L, Nakano A, Chien K . Islet1 cardiovascular progenitors: a single source for heart lineages?. Development. 2007; 135(2):193-205. DOI: 10.1242/dev.001883. View

Higuchi S, Lin Q, Wang J, Lim T, Joshi S, Anand G . Heart extracellular matrix supports cardiomyocyte differentiation of mouse embryonic stem cells. J Biosci Bioeng. 2012; 115(3):320-5. PMC: 5330950. DOI: 10.1016/j.jbiosc.2012.10.004. View

Le Guen L, Karpanen T, Schulte D, Harris N, Koltowska K, Roukens G . Ccbe1 regulates Vegfc-mediated induction of Vegfr3 signaling during embryonic lymphangiogenesis. Development. 2014; 141(6):1239-49. DOI: 10.1242/dev.100495. View

Bos F, Caunt M, Peterson-Maduro J, Planas-Paz L, Kowalski J, Karpanen T . CCBE1 is essential for mammalian lymphatic vascular development and enhances the lymphangiogenic effect of vascular endothelial growth factor-C in vivo. Circ Res. 2011; 109(5):486-91. DOI: 10.1161/CIRCRESAHA.111.250738. View

Astin J, Haggerty M, Okuda K, Le Guen L, Misa J, Tromp A . Vegfd can compensate for loss of Vegfc in zebrafish facial lymphatic sprouting. Development. 2014; 141(13):2680-90. DOI: 10.1242/dev.106591. View

Connell F, Kalidas K, Ostergaard P, Brice G, Homfray T, Roberts L . Linkage and sequence analysis indicate that CCBE1 is mutated in recessively inherited generalised lymphatic dysplasia. Hum Genet. 2009; 127(2):231-41. DOI: 10.1007/s00439-009-0766-y. View

10.

Connell F, Kalidas K, Ostergaard P, Brice G, Murday V, Mortimer P . CCBE1 mutations can cause a mild, atypical form of generalized lymphatic dysplasia but are not a common cause of non-immune hydrops fetalis. Clin Genet. 2012; 81(2):191-7. DOI: 10.1111/j.1399-0004.2011.01731.x. View

11.

Zhou B, von Gise A, Ma Q, Rivera-Feliciano J, Pu W . Nkx2-5- and Isl1-expressing cardiac progenitors contribute to proepicardium. Biochem Biophys Res Commun. 2008; 375(3):450-3. PMC: 2610421. DOI: 10.1016/j.bbrc.2008.08.044. View

12.

Burger N, Bekker M, Kok E, de Groot C, Martin J, Shou W . Increased nuchal translucency origins from abnormal lymphatic development and is independent of the presence of a cardiac defect. Prenat Diagn. 2015; 35(13):1278-86. DOI: 10.1002/pd.4687. View

13.

Alders M, Hogan B, Gjini E, Salehi F, Al-Gazali L, Hennekam E . Mutations in CCBE1 cause generalized lymph vessel dysplasia in humans. Nat Genet. 2009; 41(12):1272-4. DOI: 10.1038/ng.484. View

14.

Facucho-Oliveira J, Bento M, Belo J . Ccbe1 expression marks the cardiac and lymphatic progenitor lineages during early stages of mouse development. Int J Dev Biol. 2012; 55(10-12):1007-14. DOI: 10.1387/ijdb.113394jf. View

15.

Mommersteeg M, Dominguez J, Wiese C, Norden J, de Gier-de Vries C, Burch J . The sinus venosus progenitors separate and diversify from the first and second heart fields early in development. Cardiovasc Res. 2010; 87(1):92-101. DOI: 10.1093/cvr/cvq033. View

16.

Taylor-Weiner H, Schwarzbauer J, Engler A . Defined extracellular matrix components are necessary for definitive endoderm induction. Stem Cells. 2013; 31(10):2084-94. PMC: 3812380. DOI: 10.1002/stem.1453. View

17.

Czyz J, Wobus A . Embryonic stem cell differentiation: the role of extracellular factors. Differentiation. 2002; 68(4-5):167-74. DOI: 10.1046/j.1432-0436.2001.680404.x. View

18.

Jeltsch M, Kumar Jha S, Tvorogov D, Anisimov A, Leppanen V, Holopainen T . CCBE1 enhances lymphangiogenesis via A disintegrin and metalloprotease with thrombospondin motifs-3-mediated vascular endothelial growth factor-C activation. Circulation. 2014; 129(19):1962-71. DOI: 10.1161/CIRCULATIONAHA.113.002779. View

19.

Keller G . In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol. 1995; 7(6):862-9. DOI: 10.1016/0955-0674(95)80071-9. View

20.

Bento M, Correia E, Tavares A, Becker J, Belo J . Identification of differentially expressed genes in the heart precursor cells of the chick embryo. Gene Expr Patterns. 2011; 11(7):437-47. DOI: 10.1016/j.gep.2011.07.002. View