Heart disease often leads to cardiomyocyte death and pathological remodelling. Heart transplantation is usually the solution, although limited by donors number and restrictive inclusion criteria. These considerations prompted research into stem cell–based alternatives which however is restricted by a limited source of adult stem cells and their relative inefficient contribution to heart regeneration. Embryonic stem cells (ESC) retain great promise as an unlimited source of pluripotent progenitors for myocardial regeneration, however their therapeutics use is still impaired by the incomplete understanding of factors governing cardiomyocytes differentiation from embryonic or endogenous adult stem cells. We aimed at creating ESC-derived cardiomyocytes suitable for experimental cell transplantation therapies. We treated murine ESC with Triiodothyronine (T3), an active thyroid hormone form, and/or with anacardic acid (AA), a naturally occurring epigenetic drug that inhibits the histone acetylases (HATs), and investigated whether cardiac cell differentiation occurred and at which efficiency. To facilitate identification of differentiated cells, engineered R1 cells expressing a red fluorescent protein (RFP) gene under the NCX1 promoter, an early cardiac differentiation marker, were used. The hanging-drop embryoid body (EB) technique was used to reproduce in vitro an embryo-like architecture. ESC derived RFP-positive cardiomyocytes were collected and analysed by RT-PCR, western blot, electrophysiology and fluorescence activated cell sorting. The results show that both AA and T3 promote cardiac differentiation but with apparently distinct mechanisms: AA acts by decreasing lysine acetylation including histone H3 Lysine 9 (H3K9Ac), while T3 anticipates EBs beating increasing H3K9Ac levels. These findings underlie the possible presence of multiple epigenetically controlled signalling pathways leading to cardiomyocyte differentiation.
Role of thyroid hormones and epigenetic drugs in cardiac differentiation of mouse embryonic stem cells
RE, AGNESE;TRIMARCHI, Francesco;
2012-01-01
Abstract
Heart disease often leads to cardiomyocyte death and pathological remodelling. Heart transplantation is usually the solution, although limited by donors number and restrictive inclusion criteria. These considerations prompted research into stem cell–based alternatives which however is restricted by a limited source of adult stem cells and their relative inefficient contribution to heart regeneration. Embryonic stem cells (ESC) retain great promise as an unlimited source of pluripotent progenitors for myocardial regeneration, however their therapeutics use is still impaired by the incomplete understanding of factors governing cardiomyocytes differentiation from embryonic or endogenous adult stem cells. We aimed at creating ESC-derived cardiomyocytes suitable for experimental cell transplantation therapies. We treated murine ESC with Triiodothyronine (T3), an active thyroid hormone form, and/or with anacardic acid (AA), a naturally occurring epigenetic drug that inhibits the histone acetylases (HATs), and investigated whether cardiac cell differentiation occurred and at which efficiency. To facilitate identification of differentiated cells, engineered R1 cells expressing a red fluorescent protein (RFP) gene under the NCX1 promoter, an early cardiac differentiation marker, were used. The hanging-drop embryoid body (EB) technique was used to reproduce in vitro an embryo-like architecture. ESC derived RFP-positive cardiomyocytes were collected and analysed by RT-PCR, western blot, electrophysiology and fluorescence activated cell sorting. The results show that both AA and T3 promote cardiac differentiation but with apparently distinct mechanisms: AA acts by decreasing lysine acetylation including histone H3 Lysine 9 (H3K9Ac), while T3 anticipates EBs beating increasing H3K9Ac levels. These findings underlie the possible presence of multiple epigenetically controlled signalling pathways leading to cardiomyocyte differentiation.Pubblicazioni consigliate
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