Epigenetic control of cardiac differentiation in mouse embryonic stem cells: Role of thyroid hormone in pacemaker cell commitment
| Autor: | Re A, Granata S, Nanni S, Aiello A, Colussi C, Rossini A, Barbuti A, Mattiussi S, Trimarchi, Pontecorvi A, Gaetano C, Farsetti A |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Zdroj: | Joint Symposium_PVRI_ECCPS "Molecular Mechanism and Treatment of Heart and Lung Disease", Bad Nauheim, Germany, 27-31 January 2014 info:cnr-pdr/source/autori:Re A, Granata S, Nanni S, Aiello A, Colussi C, Rossini A, Barbuti A, Mattiussi S, Trimarchi, Pontecorvi A, Gaetano C and Farsetti A/congresso_nome:Joint Symposium_PVRI_ECCPS "Molecular Mechanism and Treatment of Heart and Lung Disease"/congresso_luogo:Bad Nauheim, Germany/congresso_data:27-31 January 2014/anno:2014/pagina_da:/pagina_a:/intervallo_pagine |
| Popis: | In order to understand whether thyroid hormone could drive ES differentiation toward cardiomyocytes, mouse ES cells (mES) were treated with Triiodothyronine (T3) in the presence or absence of anacardic acid (AA), a natural epigenetic drug inhibiting histone acetylases. To facilitate the identification of differentiated cells, engineered mES expressing red fluorescent protein (RFP) under the Na(+)/Ca(2+) exchanger gene (NCX1) promoter, an early cardiac differentiation marker, were used. The hanging-drop embryoid body (EB) technique was adopted to reproduce in vitro an embryo-like architecture. mES-derived RFP positive cardiomyocytes were collected and analysed by RT-PCR, WB, and electrophysiology. Single (AA or T3) or combined treatments (AA+T3) enhanced cardiac differentiation and increased beating areas but only T3, alone or in the presence of AA, significantly anticipated timing of EBs beating. Mechanistically, AA decreased lysine acetylation of histone and non-histone proteins while T3 did not affect Lysine acetylation level. In these conditions, RT-PCR showed decreased stemness genes expression in EBs. Moreover, AA increased expression of Nkx2.5, a cardiac differentiation key gene while T3 treatment strongly repressed miR 133a known to negatively regulate potassium voltage-gate channels (KCNA/D family), typically expressed on cardiac pacemaker cells. This finding was confirmed by patch-clamp electrophysiology showing increased spontaneous firing in support of T3-inducing pacemaker-like cell differentiation. Our data indicate T3 and AA as important controllers of multiple epigenetic signals in ES-derived cardiomyocytes and provide evidence that T3 may represent an important tool for the in vitro production of functionally specialized (pacemaker) cardiac cells suitable for therapeutic intervention. |
| Databáze: | OpenAIRE |
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