title: Enhanced efficiency of genetic programming toward cardiomyocyte creation through topographical cues.
abstract: Generation of de novo cardiomyocytes through viral over-expression of key transcription factors represents a highly promising strategy for cardiac muscle tissue regeneration. Although the feasibility of cell reprogramming has been proven possible both in vitro and in vivo, the efficiency of the process remains extremely low. Here, we report a chemical-free technique in which topographical cues, more specifically parallel microgrooves, enhance the directed differentiation of cardiac progenitors into cardiomyocyte-like cells. Using a lentivirus-mediated direct reprogramming strategy for expression of Myocardin, Tbx5, and Mef2c, we showed that the microgrooved substrate provokes an increase in histone H3 acetylation (AcH3), known to be a permissive environment for reprogramming by "stemness" factors, as well as stimulation of myocardin sumoylation, a post-translational modification essential to the transcriptional function of this key co-activator. These biochemical effects mimicked those of a pharmacological histone deacetylase inhibitor, valproic acid (VPA), and like VPA markedly augmented the expression of cardiomyocyte-specific proteins by the genetically engineered cells. No instructive effect was seen in cells unresponsive to VPA. In addition, the anisotropy resulting from parallel microgrooves induced cellular alignment, mimicking the native ventricular myocardium and augmenting sarcomere organization.
authors: Department of Materials, Imperial College London, London SW7 2AZ, United Kingdom; National Heart and Lung Institute, Imperial College London, London W12 0NN, United Kingdom; British Heart Foundation Centre of Research Excellence, Imperial College London, London W12 0NN, United Kingdom.
authors: Constant
authors: C
authors: Morez
authors: National Heart and Lung Institute, Imperial College London, London W12 0NN, United Kingdom; British Heart Foundation Centre of Research Excellence, Imperial College London, London W12 0NN, United Kingdom.
authors: Michela
authors: M
authors: Noseda
authors: National Heart and Lung Institute, Imperial College London, London W12 0NN, United Kingdom; British Heart Foundation Centre of Research Excellence, Imperial College London, London W12 0NN, United Kingdom.
authors: Marta Abreu
authors: MA
authors: Paiva
authors: National Heart and Lung Institute, Imperial College London, London W12 0NN, United Kingdom; British Heart Foundation Centre of Research Excellence, Imperial College London, London W12 0NN, United Kingdom.
authors: Elisa
authors: E
authors: Belian
authors: National Heart and Lung Institute, Imperial College London, London W12 0NN, United Kingdom; British Heart Foundation Centre of Research Excellence, Imperial College London, London W12 0NN, United Kingdom. Electronic address: m.d.schneider@imperial.ac.uk.
authors: Michael D
authors: MD
authors: Schneider
authors: Department of Materials, Imperial College London, London SW7 2AZ, United Kingdom; Department of Bioengineering, Imperial College London, London SW7 2AZ, United Kingdom; Institute for Biomedical Engineering, Imperial College London, London SW7 2AZ, United Kingdom. Electronic address: m.stevens@imperial.ac.uk.
authors: Molly M
authors: MM
authors: Stevens
mesh_terms: D000107:Acetylation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D065150:Cellular Reprogramming / Q000187:drug effects* / Q000235:genetics*; D002999:Clone Cells; D004129:Dimethylpolysiloxanes / Q000737:chemistry* / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012518:Sarcomeres / Q000187:drug effects / Q000378:metabolism; D058985:Side-Population Cells / Q000166:cytology / Q000187:drug effects; D058207:Sumoylation / Q000187:drug effects; D015534:Trans-Activators / Q000378:metabolism; D014635:Valproic Acid / Q000494:pharmacology; D000099251:Myocardin
chemical_list: D004129:Dimethylpolysiloxanes; D006657:Histones; D009687:Nuclear Proteins; D015534:Trans-Activators; D014635:Valproic Acid; D000099251:Myocardin
keywords: Cardiac tissue engineering; Cardiomyocyte; Micropatterning; Stem cell; Surface topography