Combined genetic-pharmacologic inactivation of tightly linked ADAMTS proteases in temporally specific windows uncovers distinct roles for versican proteolysis and glypican-6 in cardiac development.

Publisher: Elsevier Country of Publication: Netherlands NLM ID: 9432592 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1569-1802 (Electronic) Linking ISSN: 0945053X NLM ISO Abbreviation: Matrix Biol Subsets: MEDLINE

Publication: Amsterdam : Elsevier
Original Publication: Stuttgart ; New York : Fischer, c1994-

Versicans*/metabolism ; Versicans*/genetics ; ADAMTS5 Protein*/metabolism ; ADAMTS5 Protein*/genetics ; Proteolysis* ; ADAMTS1 Protein*/metabolism ; ADAMTS1 Protein*/genetics ; Glypicans*/metabolism ; Glypicans*/genetics ; Heart*/growth & development; Animals ; Mice ; Mice, Knockout ; Gene Expression Regulation, Developmental ; Heart Defects, Congenital/genetics ; Heart Defects, Congenital/metabolism ; Heart Defects, Congenital/pathology

Extracellular matrix remodeling mechanisms are understudied in cardiac development and congenital heart defects. We show that matrix-degrading metalloproteases ADAMTS1 and ADAMTS5, are extensively co-expressed during mouse cardiac development. The mouse mutants of each gene have mild cardiac anomalies, however, their combined genetic inactivation to elicit cooperative roles is precluded by tight gene linkage. Therefore, we coupled Adamts1 inactivation with pharmacologic ADAMTS5 blockade to uncover stage-specific cooperative roles and investigated their potential substrates in mouse cardiac development. ADAMTS5 blockade was achieved in Adamts1 null mouse embryos using an activity-blocking monoclonal antibody during distinct developmental windows spanning myocardial compaction or cardiac septation and outflow tract rotation. Synchrotron imaging, RNA in situ hybridization, immunofluorescence microscopy and electron microscopy were used to determine the impact on cardiac development and compared to Gpc6 and ADAMTS-cleavage resistant versican mutants. Mass spectrometry-based N-terminomics was used to seek relevant substrates. Combined inactivation of ADAMTS1 and ADAMTS5 prior to 12.5 days of gestation led to dramatic accumulation of versican-rich cardiac jelly and inhibited formation of compact and trabecular myocardium, which was also observed in mice with ADAMTS cleavage-resistant versican. Combined inactivation after 12.5 days impaired outflow tract development and ventricular septal closure, generating a tetralogy of Fallot-like defect. N-terminomics of combined ADAMTS knockout and control hearts identified a cleaved glypican-6 peptide only in the controls. ADAMTS1 and ADAMTS5 expression in cells was associated with specific glypican-6 cleavages. Paradoxically, combined ADAMTS1 and ADAMTS5 inactivation reduced cardiac glypican-6 and outflow tract Gpc6 transcription. Notably, Gpc6 ^-/- hearts demonstrated similar rotational defects as combined ADAMTS inactivated hearts and both had reduced hedgehog signaling. Thus, versican proteolysis in cardiac jelly at the canonical Glu ⁴⁴¹ -Ala ⁴⁴² site is cooperatively mediated by ADAMTS1 and ADAMTS5 and required for proper ventricular cardiomyogenesis, whereas, reduced glypican-6 after combined ADAMTS inactivation impairs hedgehog signaling, leading to outflow tract malrotation.
(Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Am J Physiol Cell Physiol. 2022 Apr 1;322(4):C694-C698. (PMID: 35235423)
Matrix Biol. 2007 Jul;26(6):431-41. (PMID: 17509843)
J Ultrastruct Res. 1982 Oct;81(1):1-12. (PMID: 7143533)
Nature. 2018 May;557(7705):439-445. (PMID: 29743679)
FEBS Lett. 2000 Aug 4;478(3):241-5. (PMID: 10930576)
Matrix Biol. 2021 Mar;97:40-57. (PMID: 33454424)
J Biol Chem. 1999 Sep 3;274(36):25555-63. (PMID: 10464288)
Am J Physiol Lung Cell Mol Physiol. 2020 Jan 1;318(1):L65-L75. (PMID: 31596108)
Development. 1995 Aug;121(8):2537-47. (PMID: 7671817)
J Histochem Cytochem. 1983 Jun;31(6):717-27. (PMID: 6341460)
Mamm Genome. 1999 Dec;10(12):1119-25. (PMID: 10594233)
Matrix Biol. 2010 May;29(4):304-16. (PMID: 20096780)
Matrix Biol. 2018 Oct;71-72:225-239. (PMID: 29885460)
Methods Mol Biol. 2020;2043:173-178. (PMID: 31463911)
Osteoarthritis Cartilage. 2015 Aug;23(8):1254-66. (PMID: 25800415)
J Biol Chem. 1999 Aug 13;274(33):23443-50. (PMID: 10438522)
Development. 2009 May;136(10):1761-70. (PMID: 19369393)
J Biol Chem. 2003 Oct 17;278(42):41205-12. (PMID: 12888576)
Cell Rep. 2018 Apr 10;23(2):485-498. (PMID: 29642006)
Dev Biol. 1998 Oct 1;202(1):56-66. (PMID: 9758703)
Nucleic Acids Res. 2015 Jan;43(Database issue):D290-7. (PMID: 25332401)
Nature. 2019 Feb;566(7745):490-495. (PMID: 30787436)
J Proteomics. 2021 Oct 30;249:104358. (PMID: 34450332)
Matrix Biol. 2014 Apr;35:248-52. (PMID: 24412155)
Circulation. 2018 Nov 20;138(21):e653-e711. (PMID: 30571578)
Int J Biochem Cell Biol. 2005 Sep;37(9):1838-45. (PMID: 15899586)
J Biol Chem. 2010 Jul 2;285(27):21114-25. (PMID: 20404343)
FASEB J. 2004 Apr;18(6):754-6. (PMID: 14766798)
J Biol Chem. 2001 Apr 20;276(16):13372-8. (PMID: 11278559)
Nat Methods. 2012 Jun 28;9(7):676-82. (PMID: 22743772)
Circulation. 2023 Feb 21;147(8):e93-e621. (PMID: 36695182)
J Clin Invest. 2000 Aug;106(3):349-60. (PMID: 10930438)
J Synchrotron Radiat. 2012 Nov;19(Pt 6):1029-37. (PMID: 23093766)
Dev Cell. 2009 Nov;17(5):687-98. (PMID: 19922873)
Biol Chem. 2012 Dec;393(12):1477-83. (PMID: 23667905)
Osteoarthritis Cartilage. 2022 Aug;30(8):1091-1102. (PMID: 35339693)
JCI Insight. 2018 Apr 5;3(7):. (PMID: 29618652)
J Biol Chem. 2014 Oct 3;289(40):27859-73. (PMID: 25122765)
Nat Commun. 2019 Feb 27;10(1):953. (PMID: 30814516)
Development. 2020 Jul 3;147(13):. (PMID: 32620577)
Nature. 2019 Apr;568(7751):193-197. (PMID: 30944477)
Matrix Biol. 2020 Jun;88:19-32. (PMID: 31756413)
J Cell Sci. 2014 Apr 1;127(Pt 7):1565-75. (PMID: 24496449)
J Am Coll Cardiol. 2011 Nov 15;58(21):2241-7. (PMID: 22078432)
Arterioscler Thromb Vasc Biol. 2019 Oct;39(10):2067-2081. (PMID: 31366218)
Development. 2010 Dec;137(23):4029-38. (PMID: 21041365)
Dev Biol. 2007 Oct 15;310(2):291-303. (PMID: 17822691)
Mol Cell Proteomics. 2010 May;9(5):912-27. (PMID: 20305283)
Dev Cell. 2008 Feb;14(2):298-311. (PMID: 18267097)
Nat Biotechnol. 2010 Jul;28(7):749-55. (PMID: 20562862)
Dev Biol. 2011 Sep 1;357(1):152-64. (PMID: 21749862)
Biochem J. 1995 Oct 15;311 ( Pt 2):561-5. (PMID: 7487896)
Development. 2020 Aug 14;147(15):. (PMID: 32816903)
Orphanet J Rare Dis. 2009 Jan 13;4:2. (PMID: 19144126)
Nat Biotechnol. 2010 Mar;28(3):281-8. (PMID: 20208520)
Int J Biochem Cell Biol. 2009 May;41(5):1116-26. (PMID: 18992360)
PLoS One. 2016 Oct 21;11(10):e0165079. (PMID: 27768722)
Development. 2007 Apr;134(8):1593-604. (PMID: 17344228)
Circ Res. 2019 Oct 25;125(10):884-906. (PMID: 31556812)
Dev Biol. 2005 Jul 15;283(2):357-72. (PMID: 15936751)
Gene Expr Patterns. 2009 Jun;9(5):314-23. (PMID: 19250981)
Nucleic Acids Res. 2022 Jan 7;50(D1):D543-D552. (PMID: 34723319)
Elife. 2022 May 03;11:. (PMID: 35503090)
Dev Biol. 1997 Jun 1;186(1):58-72. (PMID: 9188753)
Wiley Interdiscip Rev Dev Biol. 2013 Jul;2(4):499-530. (PMID: 24014420)
Sci Rep. 2020 Oct 7;10(1):16659. (PMID: 33028938)
J Biol Chem. 2009 Mar 27;284(13):8596-604. (PMID: 19164294)
Cell Rep. 2018 May 22;23(8):2455-2466. (PMID: 29791855)
Biomolecules. 2019 Dec 16;9(12):. (PMID: 31888141)
J Cell Biol. 2017 Sep 4;216(9):2911-2926. (PMID: 28696225)
Matrix Biol Plus. 2021 May 14;10:100064. (PMID: 34195596)
Dev Dyn. 2006 Aug;235(8):2238-47. (PMID: 16691565)

P01 HL107147 United States HL NHLBI NIH HHS; R01 HL156987 United States HL NHLBI NIH HHS; S10 OD023436 United States OD NIH HHS; S10 OD030398 United States OD NIH HHS

Keywords: Degradomics; Glypican; Metalloprotease; Proteoglycan; Terminomics; Versican

126968-45-4 (Versicans)
EC 3.4.24.- (Adamts1 protein, mouse)
EC 3.4.24.- (ADAMTS5 Protein)
EC 3.4.24.- (ADAMTS1 Protein)
EC 3.4.24.- (Adamts5 protein, mouse)
0 (Glypicans)
0 (Vcan protein, mouse)

Date Created: 20240516 Date Completed: 20240615 Latest Revision: 20241102

20250114

PMC11526477

10.1016/j.matbio.2024.05.003

38750698