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CRISPR/Cas9 editing in human pluripotent stem cell-cardiomyocytes highlights arrhythmias, hypocontractility, and energy depletion as potential therapeutic targets for hypertrophic cardiomyopathy.
Mosqueira, Diogo; Mannhardt, Ingra; Bhagwan, Jamie R; Lis-Slimak, Katarzyna; Katili, Puspita; Scott, Elizabeth; Hassan, Mustafa; Prondzynski, Maksymilian; Harmer, Stephen C; Tinker, Andrew; Smith, James G W; Carrier, Lucie; Williams, Philip M; Gaffney, Daniel; Eschenhagen, Thomas; Hansen, Arne; Denning, Chris.
Afiliação
  • Mosqueira D; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Mannhardt I; Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
  • Bhagwan JR; Partner Site Hamburg/Kiel/Lübeck, DZHK (German Center for Cardiovascular Research), Hamburg, Germany.
  • Lis-Slimak K; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Katili P; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Scott E; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Hassan M; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Prondzynski M; The Heart Centre, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Charterhouse Square, London, UK.
  • Harmer SC; Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
  • Tinker A; Partner Site Hamburg/Kiel/Lübeck, DZHK (German Center for Cardiovascular Research), Hamburg, Germany.
  • Smith JGW; The Heart Centre, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Charterhouse Square, London, UK.
  • Carrier L; The Heart Centre, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Charterhouse Square, London, UK.
  • Williams PM; Department of Stem Cell Biology, Centre of Biomolecular Sciences, University of Nottingham, UK.
  • Gaffney D; Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
  • Eschenhagen T; Partner Site Hamburg/Kiel/Lübeck, DZHK (German Center for Cardiovascular Research), Hamburg, Germany.
  • Hansen A; Molecular Therapeutics and Formulation. School of Pharmacy, University of Nottingham, UK.
  • Denning C; Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge, UK.
Eur Heart J ; 39(43): 3879-3892, 2018 11 14.
Article em En | MEDLINE | ID: mdl-29741611
ABSTRACT

Aims:

Sarcomeric gene mutations frequently underlie hypertrophic cardiomyopathy (HCM), a prevalent and complex condition leading to left ventricle thickening and heart dysfunction. We evaluated isogenic genome-edited human pluripotent stem cell-cardiomyocytes (hPSC-CM) for their validity to model, and add clarity to, HCM. Methods and

results:

CRISPR/Cas9 editing produced 11 variants of the HCM-causing mutation c.C9123T-MYH7 [(p.R453C-ß-myosin heavy chain (MHC)] in 3 independent hPSC lines. Isogenic sets were differentiated to hPSC-CMs for high-throughput, non-subjective molecular and functional assessment using 12 approaches in 2D monolayers and/or 3D engineered heart tissues. Although immature, edited hPSC-CMs exhibited the main hallmarks of HCM (hypertrophy, multi-nucleation, hypertrophic marker expression, sarcomeric disarray). Functional evaluation supported the energy depletion model due to higher metabolic respiration activity, accompanied by abnormalities in calcium handling, arrhythmias, and contraction force. Partial phenotypic rescue was achieved with ranolazine but not omecamtiv mecarbil, while RNAseq highlighted potentially novel molecular targets.

Conclusion:

Our holistic and comprehensive approach showed that energy depletion affected core cardiomyocyte functionality. The engineered R453C-ßMHC-mutation triggered compensatory responses in hPSC-CMs, causing increased ATP production and αMHC to energy-efficient ßMHC switching. We showed that pharmacological rescue of arrhythmias was possible, while MHY7 MYH6 and mutant wild-type MYH7 ratios may be diagnostic, and previously undescribed lncRNAs and gene modifiers are suggestive of new mechanisms.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Arritmias Cardíacas / Cardiomiopatia Hipertrófica / Miócitos Cardíacos / Células-Tronco Pluripotentes / Contração Miocárdica Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Arritmias Cardíacas / Cardiomiopatia Hipertrófica / Miócitos Cardíacos / Células-Tronco Pluripotentes / Contração Miocárdica Limite: Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article