title: Impact of ANKRD1 mutations associated with hypertrophic cardiomyopathy on contraction parameters of engineered heart tissue.
abstract: Hypertrophic cardiomyopathy (HCM) is a myocardial disease associated with mutations in sarcomeric genes. Three mutations were found in ANKRD1, encoding ankyrin repeat domain 1 (ANKRD1), a transcriptional co-factor located in the sarcomere. In the present study, we investigated whether expression of HCM-associated ANKRD1 mutations affects contraction parameters after gene transfer in engineered heart tissues (EHTs). EHTs were generated from neonatal rat heart cells and were transduced with adeno-associated virus encoding GFP or myc-tagged wild-type (WT) or mutant (P52A, T123M, or I280V) ANKRD1. Contraction parameters were analyzed from day 8 to day 16 of culture, and evaluated in the absence or presence of the proteasome inhibitor epoxomicin for 24 h. Under standard conditions, only WT- and T123M-ANKRD1 were correctly incorporated in the sarcomere. T123M-ANKRD1-transduced EHTs exhibited higher force and velocities of contraction and relaxation than WT- P52A- and I280V-ANKRD1 were highly unstable, not incorporated into the sarcomere, and did not induce contractile alterations. After epoxomicin treatment, P52A and I280V were both stabilized and incorporated into the sarcomere. I280V-transduced EHTs showed prolonged relaxation. These data suggest different impacts of ANKRD1 mutations on cardiomyocyte function: gain-of-function for T123M mutation under all conditions and dominant-negative effect for the I280V mutation which may come into play only when the proteasome is impaired.
authors.affiliation: Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg Eppendorf, Martinistraße 52, 20246, Hamburg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Crocini
authors.forename: Takuro
authors.initials: T
authors.lastname: Arimura
authors.forename: Silke
authors.initials: S
authors.lastname: Reischmann
authors.forename: Alexandra
authors.initials: A
authors.lastname: Eder
authors.forename: Ingke
authors.initials: I
authors.lastname: Braren
authors.forename: Arne
authors.initials: A
authors.lastname: Hansen
authors.forename: Thomas
authors.initials: T
authors.lastname: Eschenhagen
authors.forename: Akinori
authors.initials: A
authors.lastname: Kimura
authors.forename: Lucie
authors.initials: L
authors.lastname: Carrier
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D009200:Myocardial Contraction / Q000187:drug effects / Q000235:genetics*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D010641:Phenotype; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology; D051381:Rats; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: C491304:ANKRD1 protein, human; D009124:Muscle Proteins; D009687:Nuclear Proteins; D009842:Oligopeptides; D061988:Proteasome Inhibitors; D012097:Repressor Proteins; D049452:Green Fluorescent Proteins; D046988:Proteasome Endopeptidase Complex; C078846:epoxomicin