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Cited by 363Developmental Biology and Gene RegulationCongenital heart defects researchCancer Cells and Metastasis

Slug is a direct Notch target required for initiation of cardiac cushion cellularization

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Niessen, Kyle, Fu, YangXin, Chang, Linda, Hoodless, Pamela A., McFadden, Deborah, Karsan, Aly·2008·The Journal of Cell Biology
DOI 10.1083/jcb.200710067PMC PMC2483533
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Abstract

Snail family proteins are key regulators of epithelial-mesenchymal transition, but their role in endothelial-to-mesenchymal transition (EMT) is less well studied. We show that Slug, a Snail family member, is expressed by a subset of endothelial cells as well as mesenchymal cells of the atrioventricular canal and outflow tract during cardiac cushion morphogenesis. Slug deficiency results in impaired cellularization of the cardiac cushion at embryonic day (E)–9.5 but is compensated by increased Snail expression at E10.5, which restores cardiac cushion EMT. We further demonstrate that Slug, but not Snail, is directly up-regulated by Notch in endothelial cells and that Slug expression is required for Notch-mediated repression of the vascular endothelial cadherin promoter and for promoting migration of transformed endothelial cells. In contrast, transforming growth factor β (TGF-β) induces Snail but not Slug. Interestingly, activation of Notch in the context of TGF-β stimulation results in synergistic up-regulation of Snail in endothelial cells. Collectively, our data suggest that combined expression of Slug and Snail is required for EMT in cardiac cushion morphogenesis.

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  • EndMT is controlled by transcription factors including SNAI2, GATA4, and SOX9, activated in response to cues including TGFβ, WNT, and NOTCH.

    “This transdifferentiation event is controlled by various transcription factors, like SNAI214, GATA415, and SOX916,17, which are activated in response to cues from neighbouring cell types, including the TGFβ superfamily9,10, WNT18, and NOTCH19,20.”

    From Cell diversity and plasticity during atrioventricular heart valve EMTs (2023) · cites this paper as [14]

  • Both SNAI1 and SNAI2 are required for EndMT in cardiac cushion morphogenesis in mice.

    “Interestingly, both SNAI1 and SNAI2 are required for EndMT in cardiac cushion morphogenesis in mice (67).”

    From SOX9 reprograms endothelial cells by altering the chromatin landscape (2022) · cites this paper as [67]

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Research works printed but not linked: 52

  1. Armstrong, E.J., and J. Bischoff. 2004. Heart valve development: endothelial cell signaling and differentiation. Circ. Res. 95: 459–470. 15345668 PMC2810618
  2. Azhar, M., J. Schultz Jel, I. Grupp, G.W. Dorn II, P. Meneton, D.G. Molin, A.C. Gittenberger-de Groot, and T. Doetschman. 2003. Transforming growth factor beta in cardiovascular development and function. Cytokine Growth Factor Rev. 14: 391–407. 12948523 PMC3855389
  3. Bartram, U., D.G. Molin, L.J. Wisse, A. Mohamad, L.P. Sanford, T. Doetschman, C.P. Speer, R.E. Poelmann, and A.C. Gittenberger-de Groot. 2001. Double-outlet right ventricle and overriding tricuspid valve reflect disturbances of looping, myocardialization, endocardial cushion differentiation, and apoptosis in TGF-beta(2)-knockout mice. Circulation. 103: 2745–2752. 11390347
  4. Blokzijl, A., C. Dahlqvist, E. Reissmann, A. Falk, A. Moliner, U. Lendahl, and C.F. Ibanez. 2003. Cross-talk between the Notch and TGF-β signaling pathways mediated by interaction of the Notch intracellular domain with Smad3. J. Cell Biol. 163: 723–728. 14638857 PMC2173673
  5. Brown, C.B., A.S. Boyer, R.B. Runyan, and J.V. Barnett. 1999. Requirement of type III TGF-beta receptor for endocardial cell transformation in the heart. Science. 283: 2080–2082. 10092230
  6. Camenisch, T.D., D.G. Molin, A. Person, R.B. Runyan, A.C. Gittenberger-de Groot, J.A. McDonald, and S.E. Klewer. 2002a. Temporal and distinct TGFbeta ligand requirements during mouse and avian endocardial cushion morphogenesis. Dev. Biol. 248: 170–181. 12142029
  7. Camenisch, T.D., J.A. Schroeder, J. Bradley, S.E. Klewer, and J.A. McDonald. 2002b. Heart-valve mesenchyme formation is dependent on hyaluronan-augmented activation of ErbB2-ErbB3 receptors. Nat. Med. 8: 850–855. 12134143
  8. Carver, E.A., R. Jiang, Y. Lan, K.F. Oram, and T. Gridley. 2001. The mouse snail gene encodes a key regulator of the epithelial-mesenchymal transition. Mol. Cell. Biol. 21: 8184–8188. 11689706 PMC99982
  9. Chang, C.P., J.R. Neilson, J.H. Bayle, J.E. Gestwicki, A. Kuo, K. Stankunas, I.A. Graef, and G.R. Crabtree. 2004. A field of myocardial-endocardial NFAT signaling underlies heart valve morphogenesis. Cell. 118: 649–663. 15339668
  10. Crosby, C.V., P.A. Fleming, W.S. Argraves, M. Corada, L. Zanetta, E. Dejana, and C.J. Drake. 2005. VE-cadherin is not required for the formation of nascent blood vessels but acts to prevent their disassembly. Blood. 105: 2771–2776. 15604224
  11. Dahlqvist, C., A. Blokzijl, G. Chapman, A. Falk, K. Dannaeus, C.F. Ibanez, and U. Lendahl. 2003. Functional Notch signaling is required for BMP4-induced inhibition of myogenic differentiation. Development. 130: 6089–6099. 14597575
  12. Dickson, M.C., H.G. Slager, E. Duffie, C.L. Mummery, and R.J. Akhurst. 1993. RNA and protein localisations of TGF beta 2 in the early mouse embryo suggest an involvement in cardiac development. Development. 117: 625–639. 7687212
  13. Donovan, J., A. Kordylewska, Y.N. Jan, and M.F. Utset. 2002. Tetralogy of Fallot and other congenital heart defects in Hey2 mutant mice. Curr. Biol. 12: 1605–1610. 12372254
  14. Eisenberg, L.M., and R.R. Markwald. 1995. Molecular regulation of atrioventricular valvuloseptal morphogenesis. Circ. Res. 77: 1–6. 7788867
  15. Eldadah, Z.A., A. Hamosh, N.J. Biery, R.A. Montgomery, M. Duke, R. Elkins, and H.C. Dietz. 2001. Familial tetralogy of fallot caused by mutation in the jagged1 gene. Hum. Mol. Genet. 10: 163–169. 11152664
  16. Fischer, A., N. Schumacher, M. Maier, M. Sendtner, and M. Gessler. 2004. The Notch target genes Hey1 and Hey2 are required for embryonic vascular development. Genes Dev. 18: 901–911. 15107403 PMC395849
  17. Fischer, A., C. Steidl, T.U. Wagner, E. Lang, P.M. Jakob, P. Friedl, K.P. Knobeloch, and M. Gessler. 2007. Combined loss of Hey1 and HeyL causes congenital heart defects because of impaired epithelial to mesenchymal transition. Circ. Res. 100: 856–863. 17303760
  18. Garg, V., A.N. Muth, J.F. Ransom, M.K. Schluterman, R. Barnes, I.N. King, P.D. Grossfeld, and D. Srivastava. 2005. Mutations in NOTCH1 cause aortic valve disease. Nature. 437: 270–274. 16025100
  19. Hay, E.D. 2005. The mesenchymal cell, its role in the embryo, and the remarkable signaling mechanisms that create it. Dev. Dyn. 233: 706–720. 15937929
  20. Inoue, A., M.G. Seidel, W. Wu, S. Kamizono, A.A. Ferrando, R.T. Bronson, H. Iwasaki, K. Akashi, A. Morimoto, J.K. Hitzler, et al. 2002. Slug, a highly conserved zinc finger transcriptional repressor, protects hematopoietic progenitor cells from radiation-induced apoptosis in vivo. Cancer Cell. 2: 279–288. 12398892
  21. Iso, T., Y. Hamamori, and L. Kedes. 2003. Notch signaling in vascular development. Arterioscler. Thromb. Vasc. Biol. 23: 543–553. 12615665
  22. Itoh, F., S. Itoh, M.J. Goumans, G. Valdimarsdottir, T. Iso, G.P. Dotto, Y. Hamamori, L. Kedes, M. Kato, and P. ten Dijke Pt. 2004. Synergy and antagonism between Notch and BMP receptor signaling pathways in endothelial cells. EMBO J. 23: 541–551. 14739937 PMC1271801
  23. Jiang, R., Y. Lan, C.R. Norton, J.P. Sundberg, and T. Gridley. 1998. The Slug gene is not essential for mesoderm or neural crest development in mice. Dev. Biol. 198: 277–285. 9659933
  24. Karsan, A., E. Yee, and J.M. Harlan. 1996. Endothelial cell death induced by tumor necrosis factor-alpha is inhibited by the Bcl-2 family member, A1. J. Biol. Chem. 271: 27201–27204. 8910286
  25. Li, L., I.D. Krantz, Y. Deng, A. Genin, A.B. Banta, C.C. Collins, M. Qi, B.J. Trask, W.L. Kuo, J. Cochran, et al. 1997. Alagille syndrome is caused by mutations in human Jagged1, which encodes a ligand for Notch1. Nat. Genet. 16: 243–251. 9207788
  26. Loomes, K.M., L.A. Underkoffler, J. Morabito, S. Gottlieb, D.A. Piccoli, N.B. Spinner, H.S. Baldwin, and R.J. Oakey. 1999. The expression of Jagged1 in the developing mammalian heart correlates with cardiovascular disease in Alagille syndrome. Hum. Mol. Genet. 8: 2443–2449. 10556292
  27. MacKenzie, F., P. Duriez, F. Wong, M. Noseda, and A. Karsan. 2004. Notch4 inhibits endothelial apoptosis via RBP-Jkappa-dependent and -independent pathways. J. Biol. Chem. 279: 11657–11663. 14701863
  28. Masuda, S., K. Kumano, K. Shimizu, Y. Imai, M. Kurokawa, S. Ogawa, M. Miyagishi, K. Taira, H. Hirai, and S. Chiba. 2005. Notch1 oncoprotein antagonizes TGF-beta/Smad-mediated cell growth suppression via sequestration of coactivator p300. Cancer Sci. 96: 274–282. 15904468 PMC11159266
  29. Molin, D.G., U. Bartram, K. Van der Heiden, L. Van Iperen, C.P. Speer, B.P. Hierck, R.E. Poelmann, and A.C. Gittenberger-de-Groot. 2003. Expression patterns of Tgfbeta1-3 associate with myocardialisation of the outflow tract and the development of the epicardium and the fibrous heart skeleton. Dev. Dyn. 227: 431–444. 12815630
  30. Molin, D.G., M.C. DeRuiter, L.J. Wisse, M. Azhar, T. Doetschman, R.E. Poelmann, and A.C. Gittenberger-de Groot. 2002. Altered apoptosis pattern during pharyngeal arch artery remodelling is associated with aortic arch malformations in Tgfbeta2 knock-out mice. Cardiovasc. Res. 56: 312–322. 12393102
  31. Murray, S.A., and T. Gridley. 2006. Snail family genes are required for left-right asymmetry determination, but not neural crest formation, in mice. Proc. Natl. Acad. Sci. USA. 103: 10300–10304. 16801545 PMC1502452
  32. Murray, S.A., K.F. Oram, and T. Gridley. 2007. Multiple functions of Snail family genes during palate development in mice. Development. 134: 1789–1797. 17376812
  33. Nagy, A.G., M. Gertsenstein, K. Vintersten, and R. Behringer. 2003. Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory Press. 779 pp.
  34. Niessen, K., and A. Karsan. 2007. Notch signaling in the developing cardiovascular system. Am. J. Physiol. Cell Physiol. 293: C1–C11. 17376817
  35. Nieto, M.A. 2002. The snail superfamily of zinc-finger transcription factors. Nat. Rev. Mol. Cell Biol. 3: 155–166. 11994736
  36. Noseda, M., G. McLean, K. Niessen, L. Chang, I. Pollet, R. Montpetit, R. Shahidi, K. Dorovini-Zis, L. Li, B. Beckstead, et al. 2004. Notch activation results in phenotypic and functional changes consistent with endothelial-to-mesenchymal transformation. Circ. Res. 94: 910–917. 14988227
  37. Noseda, M., Y. Fu, K. Niessen, F. Wong, L. Chang, G. McLean, and A. Karsan. 2006. Smooth Muscle {alpha }-Actin Is a Direct Target of Notch/CSL. Circ. Res. 98: 1468–1470. 16741155
  38. Oda, T., A.G. Elkahloun, B.L. Pike, K. Okajima, I.D. Krantz, A. Genin, D.A. Piccoli, P.S. Meltzer, N.B. Spinner, F.S. Collins, and S.C. Chandrasekharappa. 1997. Mutations in the human Jagged1 gene are responsible for Alagille syndrome. Nat. Genet. 16: 235–242. 9207787
  39. Oka, C., T. Nakano, A. Wakeham, J.L. de la Pompa, C. Mori, T. Sakai, S. Okazaki, M. Kawaichi, K. Shiota, T.W. Mak, and T. Honjo. 1995. Disruption of the mouse RBP-J kappa gene results in early embryonic death. Development. 121: 3291–3301. 7588063
  40. Oram, K.F., E.A. Carver, and T. Gridley. 2003. Slug expression during organogenesis in mice. Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 271: 189–191. 12552634
  41. Peiro, S., M. Escriva, I. Puig, M.J. Barbera, N. Dave, N. Herranz, M.J. Larriba, M. Takkunen, C. Franci, A. Munoz, et al. 2006. Snail1 transcriptional repressor binds to its own promoter and controls its expression. Nucleic Acids Res. 34: 2077–2084. 16617148 PMC1440880
  42. Prandini, M.H., I. Dreher, S. Bouillot, S. Benkerri, T. Moll, and P. Huber. 2005. The human VE-cadherin promoter is subjected to organ-specific regulation and is activated in tumour angiogenesis. Oncogene. 24: 2992–3001. 15735710 PMC2800996
  43. Ramain, P., K. Khechumian, L. Seugnet, N. Arbogast, C. Ackermann, and P. Heitzler. 2001. Novel Notch alleles reveal a Deltex-dependent pathway repressing neural fate. Curr. Biol. 11: 1729–1738. 11719214
  44. Romano, L.A., and R.B. Runyan. 2000. Slug is an essential target of TGFbeta2 signaling in the developing chicken heart. Dev. Biol. 223: 91–102. 10864463
  45. Sanford, L.P., I. Ormsby, A.C. Gittenberger-de Groot, H. Sariola, R. Friedman, G.P. Boivin, E.L. Cardell, and T. Doetschman. 1997. TGFbeta2 knockout mice have multiple developmental defects that are non-overlapping with other TGFbeta knockout phenotypes. Development. 124: 2659–2670. 9217007 PMC3850286
  46. Sun, Y., W. Lowther, K. Kato, C. Bianco, N. Kenney, L. Strizzi, D. Raafat, M. Hirota, N.I. Khan, S. Bargo, et al. 2005. Notch4 intracellular domain binding to Smad3 and inhibition of the TGF-beta signaling. Oncogene. 24: 5365–5374. 16007227
  47. Timmerman, L.A., J. Grego-Bessa, A. Raya, E. Bertran, J.M. Perez-Pomares, J. Diez, S. Aranda, S. Palomo, F. McCormick, J.C. Izpisua-Belmonte, and J.L. de la Pompa. 2004. Notch promotes epithelial-mesenchymal transition during cardiac development and oncogenic transformation. Genes Dev. 18: 99–115. 14701881 PMC314285
  48. Van Den Akker, N.M., H. Lie-Venema, S. Maas, I. Eralp, M.C. DeRuiter, R.E. Poelmann, and A.C. Gittenberger-De Groot. 2005. Platelet-derived growth factors in the developing avian heart and maturating coronary vasculature. Dev. Dyn. 233: 1579–1588. 15973731
  49. Wang, J., S. Sridurongrit, M. Dudas, P. Thomas, A. Nagy, M.D. Schneider, J.A. Epstein, and V. Kaartinen. 2005. Atrioventricular cushion transformation is mediated by ALK2 in the developing mouse heart. Dev. Biol. 286: 299–310. 16140292 PMC1361261
  50. Wilkinson, D.G. 1992. In situ hybridization: a practical approach. Oxford University Press, New York. 224 pp.
  51. Zavadil, J., L. Cermak, N. Soto-Nieves, and E.P. Bottinger. 2004. Integration of TGF-beta/Smad and Jagged1/Notch signalling in epithelial-to-mesenchymal transition. EMBO J. 23: 1155–1165. 14976548 PMC380966
  52. Zhang, H., and A. Bradley. 1996. Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. Development. 122: 2977–2986. 8898212

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Authors
Niessen, K., Fu, Y., Chang, L., Hoodless, P. A., McFadden, D., & Karsan, A.
Affiliations
  • BC Cancer Agency (CA)
  • University of British Columbia (CA)
Corresponding author
Karsan, Aly
ORCID
Venue
The Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2008-07-28
Volume
182
Issue
2
Pages
315-325
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.200710067
PMC
PMC2483533
OpenAlex
W2160483366
Cited by
363
References
1
Citations by year
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FWCI
6.45
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Congenital heart defects research
  • Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
Keywords
Slug, snails, Notch signaling, endothelial-to-mesenchymal transition, cardiac cushions, cellularization, atrioventricular canal, outflow tract, VE-cadherin, TGF-beta, endothelial cell migration, heart development
MeSH terms
Snail Family Transcription Factors, Cadherin 5, Cell Differentiation, Cell Line, Cell Movement, Heart, Humans, Mesoderm, Promoter Regions, Genetic, Stem Cells, Transcription Factors, Antigens, CD, Cadherins, Up-Regulation, Transforming Growth Factor beta, Gene Expression Regulation, Developmental, Organogenesis, Endothelial Cells, Receptor, Notch1, Endocardial Cushions

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