Human cardiomyopathy mutations induce myocyte hyperplasia and activate hypertrophic pathways during cardiogenesis in zebrafish.
To assess the effects during cardiac development of mutations that cause human cardiomyopathy, we modeled a sarcomeric gene mutation in the embryonic zebrafish. We designed morpholino antisense oligonucleotides targeting the exon 13 splice donor site in the zebrafish cardiac troponin T (tnnt2) gene, in order to precisely recapitulate a human TNNT2 mutation that causes hypertrophic cardiomyopathy (HCM). HCM is a disease characterized by myocardial hypertrophy, myocyte and myofibrillar disarray, as well as an increased risk of sudden death. Similar to humans with HCM, the morphant zebrafish embryos displayed sarcomere disarray and there was a robust induction of myocardial hypertrophic pathways. Microarray analysis uncovered a number of shared transcriptional responses between this zebrafish model and a well-characterized mouse model of HCM. However, in contrast to adult hearts, these embryonic hearts developed cardiomyocyte hyperplasia in response to this genetic perturbation. The re-creation of a human disease-causing TNNT2 splice variant demonstrates that sarcomeric mutations can alter cardiomyocyte biology at the earliest stages of heart development with distinct effects from those observed in adult hearts despite shared transcriptional responses.
Harvard Medical School, Division of Cardiology, Massachusetts General Hospital, Boston, MA 02129, USA. jason.r.becker@vanderbilt.edu
Jason R
JR
Becker
Rahul C
RC
Deo
Andreas A
AA
Werdich
Daniela
D
Panàkovà
Shannon
S
Coy
Calum A
CA
MacRae
D017398:Alternative Splicing / Q000187:drug effects / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002312:Cardiomyopathy, Hypertrophic / Q000150:complications / Q000235:genetics* / Q000473:pathology* / Q000503:physiopathology; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006321:Heart / Q000187:drug effects / Q000196:embryology* / Q000503:physiopathology; D006352:Heart Ventricles / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D006801:Humans; D006965:Hyperplasia / Q000150:complications / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D038081:Organogenesis / Q000187:drug effects; D012518:Sarcomeres / Q000187:drug effects / Q000473:pathology; D014158:Transcription, Genetic / Q000187:drug effects; D020107:Troponin T / Q000737:chemistry / Q000235:genetics; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics
D016376:Oligonucleotides, Antisense; C545568:TNNT2 protein, human; D020107:Troponin T; D029961:Zebrafish Proteins; C480681:tnnt2a protein, zebrafish; D002118:Calcium
Bioinformatics. 2009 Nov 15;25(22):3043-4
19717575
Circ Res. 2003 Mar 7;92(4):428-36
12600890
J Clin Invest. 2007 Jul;117(7):1814-23
17607358
Cardiovasc Res. 2009 Jul 15;83(2):303-12
19293247
Exp Mol Med. 2010 Jan 31;42(1):61-8
19887893
Science. 2007 Jun 8;316(5830):1481-4
17556586
J Clin Invest. 1999 Aug;104(4):469-81
10449439
Nucleic Acids Res. 2008 Jan;36(Database issue):D768-72
17991680
Proc Natl Acad Sci U S A. 2005 Dec 13;102(50):18123-8
16332958
PLoS One. 2008 Jul 09;3(7):e2642
18612386
Physiol Rev. 2007 Apr;87(2):521-44
17429040
Hum Mol Genet. 2002 Oct 1;11(20):2499-506
12351586
J Physiol. 2004 Dec 1;561(Pt 2):515-24
15486014
Bioinformatics. 2008 Jun 15;24(12):1461-2
18441000
Nucleic Acids Res. 2010 Jan;38(Database issue):D196-203
19892828
Cardiovasc Pathol. 2001 Mar-Apr;10(2):53-7
11425598
Bioinformatics. 2003 Dec 12;19(18):2502-4
14668247
Circulation. 2006 Jul 18;114(3):216-25
16831987
Surgery. 1997 Aug;122(2):412-9
9288148
Dev Cell. 2008 Jun;14(6):962-9
18539123
Cell. 1994 Jun 3;77(5):701-12
8205619
Circ Res. 2003 Aug 8;93(3):246-53
12842917
Dev Biol. 2009 Jul 15;331(2):237-49
19427304
J Cardiovasc Electrophysiol. 2008 Jan;19(1):104-10
17916152
J Biol Chem. 2005 Feb 4;280(5):3500-6
15569689
Cell. 2001 Feb 23;104(4):557-67
11239412
Circulation. 2009 Mar 17;119(10):1398-407
19255346
Nat Genet. 2002 May;31(1):106-10
11967535
J Mol Cell Cardiol. 2003 Mar;35(3):301-9
12676545
Biochem Biophys Res Commun. 2000 Dec 29;279(3):756-61
11162425
J Biol Chem. 2000 Jan 7;275(1):624-30
10617660
J Clin Invest. 2008 Dec;118(12):3893-903
19033660
Science. 1996 May 3;272(5262):731-4
8614836
Curr Biol. 2003 Dec 16;13(24):2138-47
14680629
J Mol Endocrinol. 2008 Sep;41(3):117-24
18550896
J Clin Invest. 1998 Jun 15;101(12):2800-11
9637714