Over-expression of Fgf8 in cardiac neural crest cells leads to persistent truncus arteriosus

Abstract

During cardiogenesis, the outflow tract undergoes a complicated morphogenesis, including the re-alignment of the great blood vessels, and the separation of aorta and pulmonary trunk. The deficiency of FGF8 in the morphogenesis of outflow tract has been well studied, however, the effect of over-dosed FGF8 on the development of outflow tract remains unknown. In this study, Rosa26R-Fgf8 knock-in allele was constitutively activated by Wnt1-cre transgene in the mouse neural crest cells presumptive for the endocardial cushion of outflow tract. Surprisingly, Wnt1-cre; Rosa26R-Fgf8 mouse embryos exhibited persistent truncus arteriosus and died prior to E15.5. The cardiac neural crest cells in Wnt1-cre; Rosa26R-Fgf8 truncus arteriosus did not degenerate as in WT controls, but proliferated into a thickened endocardial cushion and then, blocked the blood outflow from cardiac chambers into the lungs, which resulted in the embryonic lethality. Although the spiral aorticopulmonary septum failed to form, the differentiaion of the endothelium and smooth muscle in the Wnt1-cre; Rosa26R-Fgf8 truncus arteriosus were impacted little. However, lineage tracing assay showed that the neural crest derived cells aggregated in the cushion layer, but failed to differentiate into the endothelium of Wnt1-cre; Rosa26R-Fgf8 truncus arteriosus. Further investigation displayed the reduced p-Akt and p-Erk immunostaining, and the decreased Bmp2 and Bmp4 transcription in the endothelium of Wnt1-cre; Rosa26R-Fgf8 truncus arteriosus. Our findings suggested that Fgf8 over-expression in cardiac neural crest impaired the formation of aorticopulmonary septum by suppressing the endothelial differentiation and stimulating the proliferation of endocardial cushion cells, which implicated a novel etiology of persistent truncus arteriosus.

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References

  1. Abu-Issa R, Smyth G, Smoak I, Yamamura K, Meyers EN (2002) Fgf8 is required for pharyngeal arch and cardiovascular development in the mouse. Development 129:4613–4625

    CAS  PubMed  Google Scholar 

  2. Creazzo TL, Godt RE, Leatherbury L, Conway SJ, Kirby ML (1998) Role of cardiac neural crest cells in cardiovascular development. Annu Rev Physiol 60:267–286

    CAS  Article  Google Scholar 

  3. Darrigrand JF, Valente M, Comai G, Martinez P, Petit M, Nishinakamura R, Osorio DS, Renault G, Marchiol C, Ribes V, Cadot B (2020) Dullard-mediated Smad1/5/8 inhibition controls mouse cardiac neural crest cells condensation and outflow tract septation. Elife 9:e50325

    CAS  Article  Google Scholar 

  4. de la Pompa JL, Epstein JA (2012) Coordinating tissue interactions: notch signaling in cardiac development and disease. Dev Cell 22:244–254

    Article  Google Scholar 

  5. Dye B, Lincoln J (2020) The Endocardium and heart valves. Cold Spring HarbPerspectBiol 12:a036723

    CAS  Article  Google Scholar 

  6. Epstein JA (2010) Franklin H. Epstein lecture. Cardiac development and implications for heart disease. N Engl J Med 363:1638–1647

    CAS  Article  Google Scholar 

  7. Frank DU, Fotheringham LK, Brewer JA, Muglia LJ, Tristani-Firouzi M, Capecchi MR, Moon AM (2002) An Fgf8 mouse mutant phenocopies human 22q11 deletion syndrome. Development 129:4591–4603

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Hutson MR, Kirby ML (2007) Model systems for the study of heart development and disease. Cardiac neural crest and conotruncal malformations. Semin Cell Dev Biol 18:101–110

    CAS  Article  Google Scholar 

  9. Hutson MR, Zhang P, Stadt HA, Sato AK, Li YX, Burch J, Creazzo TL, Kirby ML (2006) Cardiac arterial pole alignment is sensitive to FGF8 signaling in the pharynx. Dev Biol 295:486–497

    CAS  Article  Google Scholar 

  10. Ilagan R, Abu-Issa R, Brown D, Yang YP, Jiao K, Schwartz RJ, Klingensmith J, Meyers EN (2006) Fgf8 is required for anterior heart field development. Development 133:2435–2445

    CAS  Article  Google Scholar 

  11. Itoh N, Ohta H, Nakayama Y, Konishi M (2016) Roles of FGF signals in heart development, health, and disease. Front Cell Dev Biol 4:110

    PubMed  PubMed Central  Google Scholar 

  12. Jia Q, McDill BW, Li SZ, Deng C, Chang CP, Chen F (2007) Smadsignaling in the neural crest regulates cardiac outflow tract remodeling through cell autonomous and non-cell autonomous effects. Dev Biol 311:172–184

    CAS  Article  Google Scholar 

  13. Jiang X, Rowitch DH, Soriano P, McMahon AP, Sucov HM (2000) Fate of the mammalian cardiac neural crest. Development 127:1607–1616

    CAS  PubMed  Google Scholar 

  14. Keyte A, Hutson MR (2012) The neural crest in cardiac congenital anomalies. Differentiation 84:25–40

    CAS  Article  Google Scholar 

  15. Kirby ML, Waldo KL (1990) Role of neural crest in congenital heart disease. Circulation 82:332–340

    CAS  Article  Google Scholar 

  16. Kirby ML, Gale TF, Stewart DE (1983) Neural crest cells contribute to normal aorticopulmonaryseptation. Science 220:1059–1061

    CAS  Article  Google Scholar 

  17. Li J, Liu KC, Jin F, Lu MM, Epstein JA (1999) Transgenic rescue of congenital heart disease and spina bifida in Splotch mice. Development 126:2495–2503

    CAS  PubMed  Google Scholar 

  18. Li J, Chen F, Epstein JA (2000) Neural crest expression of Crerecombinase directed by the proximal Pax3 promoter in transgenic mice. Genesis 26:162–164

    CAS  Article  Google Scholar 

  19. Lin CJ, Lin CY, Chen CH, Zhou B, Chang CP (2012) Partitioning the heart: mechanisms of cardiac septation and valve development. Development 139:3277–3299

    CAS  Article  Google Scholar 

  20. Lin C, Yin Y, Bell SM, Veith GM, Chen H, Huh SH, Ornitz DM, Ma L (2013) Delineating a conserved genetic cassette promoting outgrowth of body appendages. PLoS Genet 9:e1003231

    CAS  Article  Google Scholar 

  21. Liu W, Selever J, Wang D, Lu MF, Moses KA, Schwartz RJ, Martin JF (2004) Bmp4 signaling is required for outflow-tract septation and branchial-arch artery remodeling. PNAS 101:4489–4494

    CAS  Article  Google Scholar 

  22. Liu C, Gu S, Sun C, Ye W, Song Z, Zhang Y, Chen Y (2013) FGF signaling sustains the odontogenic fate of dental mesenchyme by suppressing β-catenin signaling. Development 140:4375–4385

    CAS  Article  Google Scholar 

  23. Lopez-Sanchez C, Franco D, Bonet F, Garcia-Lopez V, Aranega A, Garcia-Martinez V (2015) Negative Fgf8-Bmp2 feed-back is regulated by miR-130 during early cardiac specification. Dev Biol 406:63–73

    CAS  Article  Google Scholar 

  24. Ma P, Gu S, Karunamuni GH, Jenkins MW, Watanabe M, Rollins AM (2016) Cardiac neural crest ablation results in early endocardial cushion and hemodynamic flow abnormalities. Am J Physiol Heart CircPhysiol 311:H1150–H1159

    Article  Google Scholar 

  25. Macatee TL, Hammond BP, Arenkiel BR, Francis L, Frank DU, Moon AM (2003) Ablation of specific expression domains reveals discrete functions of ectoderm- and endoderm-derived FGF8 during cardiovascular and pharyngeal development. Development 130:6361–6374

    CAS  Article  Google Scholar 

  26. Odelin G, Faure E, Coulpier F, Di Bonito M, Bajolle F, Studer M, Avierinos J-F, Charnay P, Topilko P, Zaffran S (2018) Krox20 defines a subpopulation of cardiac neural crest cells contributing to arterial valves and bicuspid aortic valve. Development 145:dev151944

    Article  Google Scholar 

  27. Park EJ, Ogden LA, Talbot A, Evans S, Cai C, Black BL, Frank DU, Moon AM (2006) Required, tissue-specific roles for Fgf8 in outflow tract formation and remodeling. Development 133:2419–2133

    CAS  Article  Google Scholar 

  28. Park EJ, Watanabe Y, Smyth G, Miyagawa-Tomita S, Meyers E, Klingensmith J, Camenisch T, Buckingham M, Moon AM (2008) An FGF autocrine loop initiated in second heart field mesoderm regulates morphogenesis at the arterial pole of the heart. Development. https://doi.org/10.1242/dev.025437

    Article  PubMed  PubMed Central  Google Scholar 

  29. Plein A, Fantin A, Ruhrberg C (2015) Chapter Six - Neural Crest Cells in Cardiovascular Development. Current Topics in Developmental Biology. Academic Press, Trainor PA, pp 183–200

    Google Scholar 

  30. Sakabe M, Kokubo H, Nakajima Y, Saga Y (2012) Ectopic retinoic acid signaling affects outflow tract cushion development through suppression of the myocardial Tbx2-Tgfβ2 pathway. Development 139:385–395

    CAS  Article  Google Scholar 

  31. Sato A, Scholl AM, Kuhn EN, Stadt HA, Decker JR, Pegram K, Hutson MR, Kirby ML (2011) FGF8 signaling is chemotactic for cardiac neural crest cells. Dev Biol 354:18–30

    CAS  Article  Google Scholar 

  32. Shao M, Liu C, Song Y, Ye W, He W, Yuan G, Gu S, Lin C, Ma L, Zhang Y, Tian W, Hu T, Chen Y (2015) FGF8 signaling sustains progenitor status and multipotency of cranial neural crest-derived mesenchymal cells in vivo and in vitro. J Mol Cell Biol 7:441–454

    CAS  Article  Google Scholar 

  33. Stoller JZ, Epstein JA (2005) Cardiac neural crest. Semin Cell Dev Biol 16:704–715

    CAS  Article  Google Scholar 

  34. Stottmann RW, Choi M, Mishina Y, Meyers EN, Klingensmith J (2004) BMP receptor IA is required in mammalian neural crest cells for development of the cardiac outflow tract and ventricular myocardium. Development 131:2205–2218

    CAS  Article  Google Scholar 

  35. Tang S, Snider P, Firulli AB, Conway SJ (2010) Trigenic neural crest-restricted Smad7 over-expression results in congenital craniofacial and cardiovascular defects. Dev Biol 344:233–247

    CAS  Article  Google Scholar 

  36. Waldo KL, Lo CW, Kirby ML (1999) Connexin 43 expression reflects neural crest patterns during cardiovascular development. Dev Biol 208:307–323

    CAS  Article  Google Scholar 

  37. Yamagishi H (2020) Cardiac Neural Crest. Cold Spring Harb Perspect Biol Online ahead of print.

  38. Zhou S, Wang Q, Meng Z, Peng J, Zhou Y, Song W, Wang J, Chen S, Sun K (2020) Mutations in fibroblast growth factor (FGF8) and FGF10 identified in patients with conotruncal defects. J Transl Med 18:283

    Article  Google Scholar 

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Acknowledgements

This work was supported by National Natural Science Foundation of China (Grant Numbers: 81771055 and 81970922).

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Correspondence to Jing Xiao or Chao Liu.

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Tian, A., Wang, S., Wang, H. et al. Over-expression of Fgf8 in cardiac neural crest cells leads to persistent truncus arteriosus. J Mol Histol (2021). https://doi.org/10.1007/s10735-021-09956-2

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Keyword

  • Heart development
  • Outflow tract
  • Persistent truncus arteriosus
  • FGF8
  • Neural crest