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Gene Replacement Strategies to Test the Functional Redundancy of Basic Helix–Loop–Helix Transcription Factor

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Abstract

Basic helix–loop–helix (bHLH) transcription factors control developmental decisions for a wide range of embryonic cell types. Hand1 and Hand2 are closely related bHLH proteins that control cardiac, craniofacial, and limb development. Within the developing heart, Hand1 expression becomes restricted predominantly to the left ventricle, whereas Hand2 becomes restricted predominantly to the left ventricle, for which findings have shown each Hand factor to be necessary for normal chamber formation. Forced overexpression of Hand1 throughout the early developing heart induces abnormal interventricular septal development, with resulting pathogenesis of congenital heart defects. To investigate the potential transcriptional mechanisms involved in heart morphogenesis by Hand2, this study used a replacement targeting approach to knock Hand2 into the Hand1 locus and ectopically express one copy of Hand2 within the endogenous Hand1 expression domain in the developing hearts of transgenic mice. The findings show that high-percentage Hand1 Hand2 chimeras die at birth and exhibit a range of congenital heart defects. These findings suggest that Hand factors may act via unique transcriptional mechanisms mediated by bHLH factor partner choice, supporting the notion that alterations of Hand factor stoichiometry may be as deleterious to normal heart morphogenesis as Hand factor loss of function.

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References

  1. Barnes RM, Firulli AB (2009) A Twist of insight, the role of Twist family bHLH factors in development. Int J Dev Biol 53:909–924

    Article  CAS  PubMed  Google Scholar 

  2. Bettencourt PM (2005) Clinical usefulness of B-type natriuretic peptide measurement: present and future perspectives. Heart 91:1489–1494

    Article  CAS  PubMed  Google Scholar 

  3. Bouman HG, Broekhuizen ML, Baasten AM, Gittenberger-De Groot AC, Wenink AC (1995) Spectrum of looping disturbances in stage 34 chicken hearts after retinoic acid treatment. Anatomical Record 243:101–108

    Article  CAS  PubMed  Google Scholar 

  4. Capellini TD, Di Giacomo G, Salsi V, Brendolan A, Ferretti E, Srivastava D, Zappavigna V, Selleri L (2006) Pbx1/Pbx2 requirement for distal limb patterning is mediated by the hierarchical control of Hox gene spatial distribution and Shh expression. Development 133:2263–2273

    Article  CAS  PubMed  Google Scholar 

  5. Conway SJ (1996) In situ hybridization of cell and tissue sections. Humana Press Inc, Totowa

    Google Scholar 

  6. Conway SJ, Firulli AB (2009) A bHLH code for cardiac morphogenesis. Pediatr Cardiol [Epub ahead of print]

  7. Conway SJ, Kruzynska-Frejtag A, Kneer PL, Machnicki M, Koushik SV (2003) What cardiovascular defect does my prenatal mouse mutant have, and why? Genesis 35:1–21

    Article  PubMed  Google Scholar 

  8. Cross JC, Flannery ML, Blanar MA, Steingrimsson E, Jenkins NA, Copeland NG, Rutter WJ, Werb Z (1995) Hxt encodes a basic helix–loop–helix transcription factor that regulates trophoblast cell development. Development 121:2513–2523

    CAS  PubMed  Google Scholar 

  9. Cserjesi P, Brown D, Lyons GE, Olson EN (1995) Expression of the novel basic helix–loop–helix gene eHAND in neural crest derivatives and extraembryonic membranes during mouse development. Dev Biol 170:664–678

    Article  CAS  PubMed  Google Scholar 

  10. de Bold A (1985) Atrial natriuretic factor: a hormone produced by the heart. Science 230:767–770

    Article  PubMed  Google Scholar 

  11. Eisenberg LM, Markwald RR (2004) Cellular recruitment and the development of the myocardium. Dev Biol 274:225–232

    Article  CAS  PubMed  Google Scholar 

  12. Fernandez-Teran M, Piedra ME, Rodriguez-Rey JC, Talamillo A, Ros MA (2003) Expression and regulation of eHAND during limb development. Dev Dyn 226:690–701

    Article  CAS  PubMed  Google Scholar 

  13. Field LJ, Shou W, Caldwell RL (2009) 2008 Riley Heart Center symposium on cardiac development: growth and morphogenesis of the ventricular wall. Pediatr Cardiol 30:577–579

    Article  PubMed  Google Scholar 

  14. Firulli AB (2003) A HANDful of questions: the molecular biology of the HAND subclass of basic helix–loop–helix transcription factors. Gene. 312C:27–40

    Article  Google Scholar 

  15. Firulli AB, Conway SJ (2004) Combinatorial transcriptional interaction within the cardiac neural crest: a pair of HANDs in heart formation. Birth Defects Research, Part C. Embryo Today 72:151–161

    Article  CAS  Google Scholar 

  16. Firulli AB, McFadden DG, Lin Q, Srivastava D, Olson EN (1998) Heart and extra-embryonic mesodermal defects in mouse embryos lacking the bHLH transcription factor Hand1. Nat Genet 18:266–270

    Article  CAS  PubMed  Google Scholar 

  17. Firulli BA, Hadzic DB, McDaid JR, Firulli AB (2000) The basic helix–loop–helix transcription factors dHAND and eHAND exhibit dimerization characteristics that suggest complex regulation of function. J Biol Chem 275:33567–33573

    Article  CAS  PubMed  Google Scholar 

  18. Firulli B, Howard MJ, McDaid JR, McIlreavey L, Dionne KM, Centonze V, Cserjesi P, Virshup DMa, Firulli AB (2003) PKA, PKC, and the protein phosphatase 2A influence HAND factor function: a mechanism for tissue-specific transcriptional regulation. Mol Cell 12:1225–1237

    Article  CAS  PubMed  Google Scholar 

  19. Firulli BA, Krawchuk D, Centonze VE, Virshup DE, Conway SJ, Cserjesi P, Laufer E, Firulli AB (2005) Altered Twist1 and Hand2 dimerization is associated with Saethre-Chotzen syndrome and limb abnormalities. Nat Genet 37:373–381

    Article  CAS  PubMed  Google Scholar 

  20. Habets PEMH, Moorman AFM, Clout DEW, van Roon MA, Lingbeek M, van Lohuizen M, Campione M, Christoffels VM (2002) Cooperative action of Tbx2 and Nkx2.5 inhibits ANF expression in the atrioventricular canal: implications for cardiac chamber formation. Genes Dev 16:1234–1246

    Article  CAS  PubMed  Google Scholar 

  21. Hoffman JI, Kaplan S, Liberthson RR (2004) Prevalence of congential heart disease. Am Heart J 147:425–439

    Article  PubMed  Google Scholar 

  22. Horsthuis T, Houweling AC, Habets PEMH, de Lange FJ, el Azzouzi H, Clout DEW, Moorman AFM, Christoffels VM (2008) Distinct regulation of developmental and heart disease-induced atrial natriuretic factor expression by two separate distal sequences. Circ Res 102:849–859

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  24. Koushik SV, Wang J, Rogers R, Moskophidis D, Lambert NA, Creazzo TL, Conway SJ (2001) Targeted inactivation of the sodium-calcium exchanger (Ncx1) results in the lack of a heartbeat and abnormal myofibrillar organization. FASEB J 15:1209–1211

    CAS  PubMed  Google Scholar 

  25. Lin CR, Kioussi C, O’Connell S, Briata P, Szeto D, Liu F, Izpisua-Belmonte JC, Rosenfeld MG (1999) Pitx2 regulates lung asymmetry, cardiac positioning, and pituitary and tooth morphogenesis. Nature 401:279–282

    Article  CAS  PubMed  Google Scholar 

  26. Massari ME, Murre C (2000) Helix–loop–helix proteins: regulators of transcription in eucaryotic organisms. Mol Cell Biol 20:429–440

    Article  CAS  PubMed  Google Scholar 

  27. McFadden DG, McAnally J, Richardson JA, Charite’ J, Olson EN (2002) Misexpression of dHAND induces ectopic digits in the developing limb bud in the absence of direct DNA binding. Development 129:3077–3088

    CAS  PubMed  Google Scholar 

  28. McFadden DG, Barbosa AC, Richardson JA, Schneider MD, Srivastava D, Olson EN (2005) The Hand1 and Hand2 transcription factors regulate expansion of the embryonic cardiac ventricles in a gene dosage-dependent manner. Development 132:189–201

    Article  CAS  PubMed  Google Scholar 

  29. Menick DR, Renaud L, Buchholz A, Muller JG, Zhou H, Kappler CS, Kubalak SW, Conway SJ, Xu L (2007) Regulation of Ncx1 gene expression in the normal and hypertrophic heart. Ann N Y Acad Sci 1099:195–203

    Article  CAS  PubMed  Google Scholar 

  30. Natarajan A, Yamagishi H, Ahmad F, Li D, Roberts R, Matsuoka R, Hill S, Srivastava D (2001) Human eHAND, but not dHAND, is downregulated in cardiomyopathies. J Mol Cellular Cardiol 33:1607–1614

    Article  CAS  Google Scholar 

  31. Reamon-Buettner SM, Ciribilli Y, Traverso I, Kuhls B, Inga A, Borlak J (2009) A functional genetic study identifies HAND1 mutations in septation defects of the human heart. Hum Mol Genet 18:3567–3578

    Article  CAS  PubMed  Google Scholar 

  32. Riddle RD, Johnson RL, Laufer E, Tabin C (1993) Sonic hedgehog mediates the polarizing activity of the ZPA. Cell 75:1401–1416

    Article  CAS  PubMed  Google Scholar 

  33. Riley P, Anson-Cartwright L, Cross JC (1998) The Hand1 bHLH transcription factor is essential for placentation and cardiac morphogenesis. Nat Genet 18:271–275

    Article  CAS  PubMed  Google Scholar 

  34. Rios H, Koushik SV, Wang H, Wang J, Zhou HM, Lindsley A, Rogers R, Chen Z, Maeda M, Kruzynska-Frejtag A, Feng JQ, Conway SJ (2005) Periostin null mice exhibit dwarfism, incisor enamel defects, and an early-onset periodontal disease-like phenotype. Mol Cellular Biol 25:11131–11144

    Article  CAS  Google Scholar 

  35. Ritter O, Haase H, Schulte HD, Lange PE, Morano I (1999) Remodeling of the hyperophied human myocardium by cardiac bHLH transcription factors. J Cellular Biochem 74:551–561

    Article  CAS  Google Scholar 

  36. Sakata Y, Kamei CN, Nakagami H, Bronson R, Liao JK, Chin MT (2002) Ventricular septal defect and cardiomyopathy in mice lacking the transcription factor CHF1/Hey2. Proc Natl Acad Sci USA 99:16197–16202

    Article  CAS  PubMed  Google Scholar 

  37. Snider P, Olaopa M, Firulli AB, Conway SJ (2007) Cardiovascular development and the colonizing cardiac neural crest lineage. Sci World J 7:1090–1113

    CAS  Google Scholar 

  38. Snider P, Hinton RB, Moreno-Rodriguez RA, Wang J, Rogers R, Lindsley A, Li F, Ingram DA, Menick D, Field L, Firulli AB, Molkentin JD, Markwald R, Conway SJ (2008) Periostin is required for maturation and extracellular matrix stabilization of noncardiomyocyte lineages of the heart. Circ Res 102:752–760

    Article  CAS  PubMed  Google Scholar 

  39. Srivastava D, Cserjesi P, Olson EN (1995) A subclass of bHLH proteins required for cardiac morphogenesis. Science 270:1995–1999

    Article  CAS  PubMed  Google Scholar 

  40. Srivastava D, Thomas T, Lin Q, Kirby ML, Brown D, Olson EN (1997) Regulation of cardiac mesodermal and neural crest development by the bHLH transcription factor, dHAND. Nat Genet 16:154–160

    Article  CAS  PubMed  Google Scholar 

  41. Stalmans I, Lambrechts D, De Smet F, Jansen S, Wang J, Maity S, Kneer P, von der Ohe M, Swillen A, Maes C, Gewillig M, Molin DGM, Hellings P, Boetel T, Haardt M, Compernolle V, Dewerchin M, Plaisance S, Vlietinck R, Emanuel B, Gittenberger-de Groot AC, Scambler P, Morrow B, Driscol DA, Moons L, Esguerra CV, Carmeliet G, Behn-Krappa A, Devriendt K, Collen D, Conway SJ, Carmeliet P (2003) VEGF: a modifier of the del22q11 (DiGeorge) syndrome? Nat Med 9:173–182

    Article  CAS  PubMed  Google Scholar 

  42. Thattaliyath BD, Firulli BA, Firulli AB (2002) The basic-helix–loop–helix transcription factor HAND2 directly regulates transcription of the atrial naturetic peptide gene. J Mol Cell Cardiol 34:1325–1344

    Article  Google Scholar 

  43. Thattaliyath BD, Livi CB, Steinhelper ME, Toney GM, Firulli AB (2002) HAND1 and HAND2 are expressed in the adult rodent heart and are modulated during cardiac hypertrophy. Biochem Biophys Res Com 297:870–875

    Article  CAS  PubMed  Google Scholar 

  44. Thomas T, Yamagishi H, Overbeek PA, Olson EN, Srivastava D (1998) The bHLH factors, dHAND and eHAND, specify pulmonary and systemic cardiac ventricles independent of left-right sidedness. Dev Biol 196:228–236

    Article  CAS  PubMed  Google Scholar 

  45. Tickle C (1981) The number of polarizing region cells required to specify additional digits in the developing chick wing. Nature 289:295–298

    Article  CAS  PubMed  Google Scholar 

  46. Togi K, Kawamoto T, Yamauchi R, Yoshida Y, Kita T, Tanaka M (2004) Role of Hand1/eHAND in the dorsoventral patterning and interventricular septum formation in the embryonic heart. Mol Cell Biol 24:4627–4635

    Article  CAS  PubMed  Google Scholar 

  47. Vincentz JW, BArnes RM, Rodgers R, Firulli BA, Conway SJ, Firulli AB (2008) An absence of Twist1 results in aberrant cardiac neural crest morphogenesis. Dev Biol 320:131–139

    Article  CAS  PubMed  Google Scholar 

  48. Yamagishi H, Olson EN, Srivastava D (2000) The basic helix–loop–helix transcription factor, dHAND, is required for vascular development. J Clin Invest 105:261–270

    Article  CAS  PubMed  Google Scholar 

  49. Yasue H, Yoshimura M, Sumida H, Kikuta K, Kugiyama K, Jougasaki M, Ogawa H, Okumura K, Mukoyama M, Nakao K (1994) Localization and mechanism of secretion of B-type natriuretic peptide in comparison with those of A-type natriuretic peptide in normal subjects and patients with heart failure. Circulation 90:195–203

    CAS  PubMed  Google Scholar 

  50. Zeller R, Lopez-Rios J, Zuniga A (2009) Vertebrate limb bud development: moving towards integrative analysis of organogenesis. Nat Rev Genet 10:845–858

    Article  CAS  PubMed  Google Scholar 

  51. Zhu J, Nakamura E, Nguyen M-T, Bao X, Akiyama H, Mackem S (2008) Uncoupling sonic hedgehog control of pattern and expansion of the developing limb bud. Dev Cell 14:624–632

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported in part by the Riley Children’s Foundation, the Indiana University Department of Pediatrics (Cardiology), and NIH P01 HL085098 (to SJC and ABF).

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Authors and Affiliations

  1. Riley Heart Research Center, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 West Walnut Street, Room R4 W379, Indianapolis, IN, 46202, USA

    Anthony B. Firulli, Beth A. Firulli, Jian Wang, Rhonda H. Rogers & Simon J. Conway

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  1. Anthony B. Firulli
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  2. Beth A. Firulli
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  4. Rhonda H. Rogers
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  5. Simon J. Conway
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Correspondence to Simon J. Conway.

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Firulli, A.B., Firulli, B.A., Wang, J. et al. Gene Replacement Strategies to Test the Functional Redundancy of Basic Helix–Loop–Helix Transcription Factor. Pediatr Cardiol 31, 438–448 (2010). https://doi.org/10.1007/s00246-010-9669-x

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