Involvement of the ubiquitin-proteasome system in the pathogenesis of familial hypertrophic cardiomyopathy
Principal investigators
Lucie Carrier, Thomas Eschenhagen,
Institute of Experimental and Clinical Pharmacology and Toxicology, UKE
Co-workers partly involved
Saskia Schlossarek
Giulia Mearini
Karim Sultan
Nessan Hergesell
Lutz Pohlmann
Elisabeth Krämer
Denise Juhr
Contact:
l.carrier@uke.uni-hamburg.de
Objectives
To investigate the involvement of the UPS and its various components in the development of familial hypertrophic cardiomyopathy
Questions
- Do cMyBP-C mutants impair the UPS in vivo?
- Which UPS components regulate cMyBP-C stability and are rate-limiting in UPS inhibition?
- Does UPS inhibition by truncated cMyBP-C result in myocyte hypertrophy?
- Are the UPS components affected in various forms of cardiac hypertrophy?
- What is the impact of chronic pharmacological UPS inhibition in vivo?
Material
- cMyBPC mice (KO, KI and TG)
- UbG76V-GFP mice – GFPdgn mice
- MuRF1 KO mice
- Atrogin-1 TG mice
- Recombinant adenovirus (cMyBPC, atrogin-1, UbG76V-DsRed)
Methods
- Targeted and additional transgenesis
- Cardiac function analysis (Echocardiography, Millar-tip catheter)
- Molecular biology (qRT-PCR, Western-blot,...)
- Cell biology and biochemistry (Neonatal and adult cardiac myoyctes, fluorescence and confocal microscopy, proteasome activity assays, pulse-chase analysis, Immuno-precipitation…)
- Cell physiology (contractility and Ca2+ transient measurements in isolated intact mouse cardiac myocytes)
Selected references
- Flavigny J, Souchet M, Sébillon P, Berrebi-Bertrand I, Hainque B, Mallet A, Bril A, Schwartz K, Carrier L (1999) COOH-terminal truncated cardiac myosin binding protein C mutants resulting from familial hypertrophic cardiomyopathy mutations exhibit altered expression and/or incorporation in fetal rat cardiomyocytes. J Mol Biol 294:443-456
- Carrier L, Knoell R, Vignier N, Keller D, Ambroisine ML, Bausero P, Prudhon B, Isnard R, Fiszman M, Ross J, Schwartz K, Chien KR (2004) Asymmetric septal hypertrophy in heterozygous cMyBP-C null mice. Cardiovasc Res 63: 293-304
- Sarikas A / Carrier L, Schenke C, Doll D, Flavigny J, Lindenberg K, Eschenhagen T, Zolk O (2005) Impairment of the ubiquitin-proteasome system by truncated cardiac myosin binding protein C mutants. Cardiovasc Res 66:33-44 (Associated Editorial)
- Guse AH, Gu X, Zhang L, Webe, K, Krämer E, Yang Z, Jin H, Carrier L, Zhang L (2005) A minimal structural analogue of cyclic ADP-ribose: synthesis and calcium release activity in mammalian cells. J Biol Chem 280:15952-15959
- Cazorla O, Szilagyi S, Vignier N, Salazar G, Krämer E, Vassort G, Carrier L, Lacampagne A. (2006) Length and PKA modulations of myocytes in cardiac myosin-binding protein C deficient mice. Cardiovasc Res, 69:370-380 (Associated Editorial)
