Sci Rep. 2022 Jan 07;12(1):8. doi: 10.1038/s41598-021-03795-7.

Decrease of Pdzrn3 is required for heart maturation and protects against heart failure.

Scientific reports

Mathieu Pernot, Béatrice Jaspard-Vinassa, Alice Abelanet, Sebastien Rubin, Isabelle Forfar, Sylvie Jeanningros, Laura Cetran, Murielle Han-Yee Yu, Elise Balse, Stéphane Hatem, Pascale Dufourcq, Thierry Couffinhal, Cécile Duplàa

PMID: 34996942 DOI: 10.1038/s41598-021-03795-7

Abstract

Heart failure is the final common stage of most cardiopathies. Cardiomyocytes (CM) connect with others via their extremities by intercalated disk protein complexes. This planar and directional organization of myocytes is crucial for mechanical coupling and anisotropic conduction of the electric signal in the heart. One of the hallmarks of heart failure is alterations in the contact sites between CM. Yet no factor on its own is known to coordinate CM polarized organization. We have previously shown that PDZRN3, an ubiquitine ligase E3 expressed in various tissues including the heart, mediates a branch of the Planar cell polarity (PCP) signaling involved in tissue patterning, instructing cell polarity and cell polar organization within a tissue. PDZRN3 is expressed in the embryonic mouse heart then its expression dropped significantly postnatally corresponding with heart maturation and CM polarized elongation. A moderate CM overexpression of Pdzrn3 (Pdzrn3 OE) during the first week of life, induced a severe eccentric hypertrophic phenotype with heart failure. In models of pressure-overload stress heart failure, CM-specific Pdzrn3 knockout showed complete protection against degradation of heart function. We reported that Pdzrn3 signaling induced PKC ζ expression, c-Jun nuclear translocation and a reduced nuclear ß catenin level, consistent markers of the planar non-canonical Wnt signaling in CM. We then show that subcellular localization (intercalated disk) of junction proteins as Cx43, ZO1 and Desmoglein 2 was altered in Pdzrn3 OE mice, which provides a molecular explanation for impaired CM polarization in these mice. Our results reveal a novel signaling pathway that controls a genetic program essential for heart maturation and maintenance of overall geometry, as well as the contractile function of CM, and implicates PDZRN3 as a potential therapeutic target for the prevention of human heart failure.

© 2022. The Author(s).

References

1. Balse, E. et al. Dynamic of ion channel expression at the plasma membrane of cardiomyocytes. Physiol. Rev. 92, 1317–1358 (2012). - PubMed
2. Le Garrec, J.-F. et al. Quantitative analysis of polarity in 3D reveals local cell coordination in the embryonic mouse heart. Development 140, 395–404 (2013). - PubMed
3. Yang, Y. & Mlodzik, M. Wnt-frizzled/planar cell polarity signaling: Cellular orientation by facing the wind (Wnt). Annu. Rev. Cell Dev. Biol. 31, 623–646 (2015). - PubMed
4. Pandur, P., Lasche, M., Eisenberg, L. M. & Kuhl, M. Wnt-11 activation of a non-canonical Wnt signalling pathway is required for cardiogenesis. Nature 418, 636–641 (2002). - PubMed
5. Nagy, I. et al. Wnt-11 signalling controls ventricular myocardium development by patterning N-cadherin and beta-catenin expression. Cardiovasc. Res. 85, 100–109 (2010). - PubMed
6. Henderson, D. J. et al. Cardiovascular defects associated with abnormalities in midline development in the Loop-tail mouse mutant. Circ. Res. 89, 6–12 (2001). - PubMed
7. Leung, C., Lu, X., Liu, M. & Feng, Q. Rac1 signaling is critical to cardiomyocyte polarity and embryonic heart development. J. Am. Heart Assoc. 3, e001271 (2014). - PubMed
8. Sewduth, R. N. et al. The ubiquitin ligase PDZRN3 is required for vascular morphogenesis through Wnt/planar cell polarity signalling. Nat. Commun. 5, 4832 (2014). - PubMed
9. Sewduth, R. N. et al. PDZRN3 destabilizes endothelial cell-cell junctions through a PKCzeta-containing polarity complex to increase vascular permeability. Sci. Signaling 10, 464 (2017). - PubMed
10. Ho, H. et al. Mechanisms of Wnt5a-ror signaling in development and disease. FASEB J. 34, 1–1 (2020). - PubMed
11. Chhabra, E. S. & Higgs, H. N. The many faces of actin: Matching assembly factors with cellular structures. Nat. Cell Biol. 9, 1110–1121 (2007). - PubMed
12. Gottardi, C. J., Arpin, M., Fanning, A. S. & Louvard, D. The junction-associated protein, zonula occludens-1, localizes to the nucleus before the maturation and during the remodeling of cell-cell contacts. Proc. Natl. Acad. Sci. 93, 10779–10784 (1996). - PubMed
13. Yoshida, M. et al. Alterations in adhesion junction precede gap junction remodelling during the development of heart failure in cardiomyopathic hamsters. Cardiovasc. Res. 92, 95–105 (2011). - PubMed
14. Krusche, C. A. et al. Desmoglein 2 mutant mice develop cardiac fibrosis and dilation. Basic Res. Cardiol. 106, 617–633 (2011). - PubMed
15. Schlipp, A. et al. Desmoglein-2 interaction is crucial for cardiomyocyte cohesion and function. Cardiovasc. Res. 104, 245–257 (2014). - PubMed
16. Porrello, E. R. & Olson, E. N. A neonatal blueprint for cardiac regeneration. Stem Cell Res. 13, 556–570 (2014). - PubMed
17. Heineke, J. & Molkentin, J. D. Regulation of cardiac hypertrophy by intracellular signalling pathways. Nat. Rev. Mol. Cell Biol. 7, 589–600 (2006). - PubMed
18. Hoshijima, M. & Chien, K. R. Mixed signals in heart failure: Cancer rules. J. Clin. Invest. 109, 849–855 (2002). - PubMed
19. Nomura, S. et al. Cardiomyocyte gene programs encoding morphological and functional signatures in cardiac hypertrophy and failure. Nat. Commun. 9, 4435 (2018). - PubMed
20. Gessert, S. & Kühl, M. The multiple phases and faces of wnt signaling during cardiac differentiation and development. Circ. Res. 107, 186–199 (2010). - PubMed
21. Jeong, M.-H. et al. Cdon deficiency causes cardiac remodeling through hyperactivation of WNT/β-catenin signaling. Proc. Natl. Acad. Sci. USA 114, E1345–E1354 (2017). - PubMed
22. Mazzotta, S. et al. Distinctive roles of canonical and noncanonical wnt signaling in human embryonic cardiomyocyte development. Stem Cell Rep. 7, 764–776 (2016). - PubMed
23. Malekar, P. et al. Wnt signaling is critical for maladaptive cardiac hypertrophy and accelerates myocardial remodeling. Hypertension 55, 939–945 (2010). - PubMed
24. Haq, S. et al. Stabilization of beta-catenin by a Wnt-independent mechanism regulates cardiomyocyte growth. Proc. Natl. Acad. Sci. USA. 100, 4610–4615 (2003). - PubMed
25. Blankesteijn, W. M., van de Schans, V. A., ter Horst, P. & Smits, J. F. The Wnt/frizzled/GSK-3 beta pathway: S novel therapeutic target for cardiac hypertrophy. Trends Pharmacol. Sci. 29, 175–180 (2008). - PubMed
26. Qu, J. et al. Cardiac-specific haploinsufficiency of beta-catenin attenuates cardiac hypertrophy but enhances fetal gene expression in response to aortic constriction. J. Mol. Cell. Cardiol. 43, 319–326 (2007). - PubMed
27. Petrich, B. G. et al. Targeted activation of c-Jun N-terminal kinase in vivo induces restrictive cardiomyopathy and conduction defects. J. Biol. Chem. 279, 15330–15338 (2004). - PubMed
28. Abdul-Ghani, M. et al. Wnt11 promotes cardiomyocyte development by caspase-mediated suppression of canonical wnt signals. Mol. Cell. Biol. 31, 163–178 (2011). - PubMed
29. Toyofuku, T. et al. Direct association of the gap junction protein connexin-43 with ZO-1 in cardiac myocytes. J. Biol. Chem. 273, 12725–12731 (1998). - PubMed
30. Barker, R. J., Price, R. L. & Gourdie, R. G. Increased association of ZO-1 with connexin43 during remodeling of cardiac gap junctions. Circ. Res. 90, 317–324 (2002). - PubMed
31. Laing, J. G., Tadros, P. N., Westphale, E. M. & Beyer, E. C. Degradation of connexin43 gap junctions involves both the proteasome and the lysosome. Exp. Cell Res. 236, 482–492 (1997). - PubMed
32. Danik, S. B. et al. Modulation of cardiac gap junction expression and arrhythmic susceptibility. Circ. Res. 95, 1035–1041 (2004). - PubMed
33. Gutstein, D. E. et al. Conduction slowing and sudden arrhythmic death in mice with cardiac-restricted inactivation of connexin43. Circ. Res. 88, 333–339 (2001). - PubMed
34. van Rijen, H. V. M. et al. Slow conduction and enhanced anisotropy increase the propensity for ventricular tachyarrhythmias in adult mice with induced deletion of connexin43. Circulation 109, 1048–1055 (2004). - PubMed
35. Saffitz, J. E. Arrhythmogenic cardiomyopathy and abnormalities of cell-to-cell coupling. Heart Rhythm 6, S62–S65 (2009). - PubMed
36. Eshkind, L. Loss of desmoglein 2 suggests essential functions for early embryonic development and proliferation of embryonal stem cells. Eur. J. Cell Biol. 81, 592–598 (2002). - PubMed
37. Maron, B. J. et al. Contemporary definitions and classification of the cardiomyopathies: An American Heart Association Scientific Statement From the Council on Clinical Cardiology, Heart Failure and Transplantation Committee; Quality of Care and Outcomes Research and Functional Genomics and Translational Biology Interdisciplinary Working Groups; and Council on Epidemiology and Prevention. Circulation 113, 1807–1816 (2006). - PubMed
38. Debus, J. D. et al. In vitro analysis of arrhythmogenic cardiomyopathy associated desmoglein-2 (DSG2) mutations reveals diverse glycosylation patterns. J. Mol. Cell. Cardiol. 129, 303–313 (2019). - PubMed
39. Gutstein, D. E. The organization of adherens junctions and desmosomes at the cardiac intercalated disc is independent of gap junctions. J. Cell Sci. 116, 875–885 (2003). - PubMed
40. Nekrasova, O. E. et al. Desmosomal cadherins utilize distinct kinesins for assembly into desmosomes. J. Cell Biol. 195, 1185–1203 (2011). - PubMed
41. Ehler, E. et al. Alterations at the intercalated disk associated with the absence of muscle LIM protein. J. Cell Biol. 153, 763–772 (2001). - PubMed
42. Ferreira-Cornwell, M. C. et al. Remodeling the intercalated disc leads to cardiomyopathy in mice misexpressing cadherins in the heart. J. Cell Sci. 115, 1623–1634 (2002). - PubMed
43. Phillips, H. M. et al. Disruption of planar cell polarity signaling results in congenital heart defects and cardiomyopathy attributable to early cardiomyocyte disorganization. Circ. Res. 101, 137–145 (2007). - PubMed
44. Bruce, A. F., Rothery, S., Dupont, E. & Severs, N. J. Gap junction remodelling in human heart failure is associated with increased interaction of connexin43 with ZO-1. Cardiovasc. Res. 77, 757–765 (2008). - PubMed
45. Laing, J. G., Saffitz, J. E., Steinberg, T. H. & Yamada, K. A. Diminished zonula occludens-1 expression in the failing human heart. Cardiovasc. Pathol. 16, 159–164 (2007). - PubMed
46. Kostin, S. Zonula occludens-1 and connexin 43 expression in the failing human heart. J. Cell Mol. Med. 11, 892–895 (2007). - PubMed
47. Sepp, R., Severs, N. J. & Gourdie, R. G. Altered patterns of cardiac intercellular junction distribution in hypertrophic cardiomyopathy. Heart 76, 412–417 (1996). - PubMed
48. Severs, N. J., Bruce, A. F., Dupont, E. & Rothery, S. Remodelling of gap junctions and connexin expression in diseased myocardium. Cardiovasc. Res. 80, 9–19 (2008). - PubMed
49. Hong, T.-T. et al. BIN1 is reduced and Cav12 trafficking is impaired in human failing cardiomyocytes. Heart Rhythm 9, 812–820 (2012). - PubMed
50. Smyth, J. W. et al. Limited forward trafficking of connexin 43 reduces cell-cell coupling in stressed human and mouse myocardium. J. Clin. Invest. 120, 266–279 (2010). - PubMed
51. Sohal, D. S. et al. Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible cre protein. Circ. Res. 89, 20–25 (2001). - PubMed
52. https://fr.freedownloadmanager.org/Windows-PC/LabChart.html . - PubMed
53. https://imagej.net/Fiji . Accessed 24 Jan 2020. - PubMed
54. https://www.bruker.com/.../infrared-and-raman/opus-spectroscopy-software/downloads.html . - PubMed
55. Hackett, M. J. et al. Subcellular biochemical investigation of Purkinje neurons using synchrotron radiation Fourier transform infrared spectroscopic imaging with a focal plane array detector. ACS Chem. Neurosci. 4, 1071–1080 (2013). - PubMed
56. https://www.graphpad.com/scientific-software/prism/ (2020). - PubMed

Publication Types

• Journal Article

Grant support

• ANR -16-CE17-0001-01/Agence National de la Recherche ANR -16-CE17-0001-01