Contractile abnormalities and altered drug response in engineered heart tissue from Mybpc3-targeted knock-in mice
- PMID: 23896226
- DOI: 10.1016/j.yjmcc.2013.07.011
Contractile abnormalities and altered drug response in engineered heart tissue from Mybpc3-targeted knock-in mice
Abstract
Myosin-binding protein C (Mybpc3)-targeted knock-in mice (KI) recapitulate typical aspects of human hypertrophic cardiomyopathy. We evaluated whether these functional alterations can be reproduced in engineered heart tissue (EHT) and yield novel mechanistic information on the function of cMyBP-C. EHTs were generated from cardiac cells of neonatal KI, heterozygous (HET) or wild-type controls (WT) and developed without apparent morphological differences. KI had 70% and HET 20% lower total cMyBP-C levels than WT, accompanied by elevated fetal gene expression. Under standard culture conditions and spontaneous beating, KI EHTs showed more frequent burst beating than WT and occasional tetanic contractions (14/96). Under electrical stimulation (6Hz, 37°C) KI EHTs exhibited shorter contraction and relaxation times and a twofold higher sensitivity to external [Ca(2+)]. Accordingly, the sensitivity to verapamil was 4-fold lower and the response to isoprenaline or the Ca(2+) sensitizer EMD 57033 2- to 4-fold smaller. The loss of EMD effect was verified in 6-week-old KI mice in vivo. HET EHTs were apparently normal under basal conditions, but showed similarly altered contractile responses to [Ca(2+)], verapamil, isoprenaline and EMD. In contrast, drug-induced changes in intracellular Ca(2+) transients (Fura-2) were essentially normal. In conclusion, the present findings in auxotonically contracting EHTs support the idea that cMyBP-C's normal role is to suppress force generation at low intracellular Ca(2+) and stabilize the power-stroke step of the cross bridge cycle. Pharmacological testing in EHT unmasked a disease phenotype in HET. The altered drug response may be clinically relevant.
Keywords: ATPase, Ca(2+) transporting, cardiac mRNA; Acta1; Alpha skeletal actin protein; Atp2a2; BPM; Beats per minute; Cardiac myosin-binding protein C; Cardiac myosin-binding protein C (cMyBP-C); Contraction time (spontaneous beating); Diastolic left ventricular internal diameter; Disease modeling; EC(50); EHT; EMD; EMD 57033; Engineered heart tissue; Engineered heart tissue (EHT); HCM; HET; Half maximal effective concentration; Half maximal inhibitory concentration; Heterozygous Mybpc3-targeted knock-in mice; Homozygous Mybpc3-targeted knock-in mice; Human cardiac myosin-binding protein C gene; Hypertrophic cardiomyopathy; IC(50); ISO; Induced pluripotent stem cells; Isoprenaline; KI; LVIDd; LVIDs; LVMBW; Left ventricular mass/body weight ratio; MYBPC3; Mouse cardiac myosin-binding protein C gene or mRNA; Mybpc3; Myh6; Myh7; Myofilament Ca(2+) sensitivity; Relaxation time (spontaneous beating); Slc8a1; Sodium calcium exchanger; Systolic left ventricular internal diameter; T1; T2; TTP; TTP50; TTR50; TTR90; Time to 100% of maximal twitch force; Time to 50% of maximal twitch force; Time to 50% of relaxation; Time to 90% of relaxation; cMyBP-C; iPSC; α-myosin heavy chain, α-MHC gene or mRNA; α-skAct; α-skeletal actin gene or mRNA; β-myosin heavy chain, β-MHC gene or mRNA.
© 2013. Published by Elsevier Ltd. All rights reserved.
Similar articles
-
Comparison of the effects of a truncating and a missense MYBPC3 mutation on contractile parameters of engineered heart tissue.J Mol Cell Cardiol. 2016 Aug;97:82-92. doi: 10.1016/j.yjmcc.2016.03.003. Epub 2016 Apr 22. J Mol Cell Cardiol. 2016. PMID: 27108529
-
Translational investigation of electrophysiology in hypertrophic cardiomyopathy.J Mol Cell Cardiol. 2021 Aug;157:77-89. doi: 10.1016/j.yjmcc.2021.04.009. Epub 2021 May 3. J Mol Cell Cardiol. 2021. PMID: 33957110 Free PMC article.
-
Phosphomimetic cardiac myosin-binding protein C partially rescues a cardiomyopathy phenotype in murine engineered heart tissue.Sci Rep. 2019 Dec 3;9(1):18152. doi: 10.1038/s41598-019-54665-2. Sci Rep. 2019. PMID: 31796859 Free PMC article.
-
Cardiac myosin-binding protein C: hypertrophic cardiomyopathy mutations and structure-function relationships.Pflugers Arch. 2014 Feb;466(2):201-6. doi: 10.1007/s00424-013-1400-3. Epub 2013 Nov 17. Pflugers Arch. 2014. PMID: 24240729 Review.
-
The mechanics of the heart: zooming in on hypertrophic cardiomyopathy and cMyBP-C.FEBS Lett. 2022 Mar;596(6):703-746. doi: 10.1002/1873-3468.14301. Epub 2022 Feb 28. FEBS Lett. 2022. PMID: 35224729 Review.
Cited by
-
Investigating the role of uncoupling of troponin I phosphorylation from changes in myofibrillar Ca(2+)-sensitivity in the pathogenesis of cardiomyopathy.Front Physiol. 2014 Aug 25;5:315. doi: 10.3389/fphys.2014.00315. eCollection 2014. Front Physiol. 2014. PMID: 25202278 Free PMC article. Review.
-
Cardiac myosin-binding protein C (MYBPC3) in cardiac pathophysiology.Gene. 2015 Dec 1;573(2):188-97. doi: 10.1016/j.gene.2015.09.008. Epub 2015 Sep 8. Gene. 2015. PMID: 26358504 Free PMC article. Review.
-
Cardiovascular Organ-on-a-Chip Platforms for Drug Discovery and Development.Appl In Vitro Toxicol. 2016 Jun 1;2(2):82-96. doi: 10.1089/aivt.2016.0002. Appl In Vitro Toxicol. 2016. PMID: 28971113 Free PMC article. Review.
-
cMyBP-C ablation in human engineered cardiac tissue causes progressive Ca2+-handling abnormalities.J Gen Physiol. 2023 Apr 3;155(4):e202213204. doi: 10.1085/jgp.202213204. Epub 2023 Mar 9. J Gen Physiol. 2023. PMID: 36893011 Free PMC article.
-
Sexual dimorphic response to exercise in hypertrophic cardiomyopathy-associated MYBPC3-targeted knock-in mice.Pflugers Arch. 2015 Jun;467(6):1303-17. doi: 10.1007/s00424-014-1570-7. Epub 2014 Jul 11. Pflugers Arch. 2015. PMID: 25010737
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
Research Materials
Miscellaneous