- Email: [email protected]
0113 : Epicardial progenitors are source of adipocyte in human atria
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Archives of Cardiovascular Diseases Supplements (2016) 8, 255-257
Topic 28 – Cellular and gene therapy, stem cells April 08th, Friday 2016
0464 Selenoprotein T gene therapy with adeno-associated virus-9 improves cardiac function/remodeling in rats with heart failure Inès Boukhalfa * (1), Orianne Henri (1), Jean-Paul Henry (1), Youssef Anouar (2), Vincent Richard (1), Sahil Adriouch (3), Paul Mulder (1) (1) Université de Rouen, Rouen, France – (2) Université de Rouen, INSERM U982, Rouen, France – (3) Université de Rouen, INSERM U905, Rouen, France * Corresponding author: [email protected] (Inès Boukhalfa) Background Selenoprotein T (SelT) is a thioredoxin-like protein, which is abundantly but transiently expressed in the heart only during the embryonic development. Although SelT seems to play a limited role in adulthood, preliminary data from our laboratory showed that myocardial infarction increases SelT protein expression. This suggests that SelT may play a yet unrevealed role in cardiovascular diseases. Our studies have proven that SelT supplementation, using osmotic minipump delivery system, clearly improves cardiac remodeling and function, cardiac perfusion and decreases cardiac inflammation pathways after experimental heart failure (HF). Thus, we sought to investigate the cardiac effects of SelT over-expression by administration of recombinant adeno-associated virus 9 (rAAV9) in HF. Methods Wistar rats received a single intravenous injection of either rAAV9-SelT or saline serum one week after coronary artery ligation and left ventricular function/remodeling was determined 3 months after ligation by echocardiography. Results rAAV9-SelT (1.1011 virus-genome copies) resulted in an increased cardiac SelT expression (+150 %, p<0.05).This SelT over-expression reduced HF induced increase in left ventricular diastolic (sham: 6.14±0.16; HF: 10.83±0.36, p<0.05 vs. sham; HF+rAAV9-SelT: 9.92±0.21 mm, p<0.05 vs. HF) and systolic diameters (sham: 3.08±0.17; HF, 9.13±0.35, p<0.05 vs. sham; CAL+rAAV9-SelT: 8.14±0.27 mm, p<0.05 vs HF). Simultaneously, SelT improved stroke volume (sham: 0.49±0.02; HF: 0.41±0.01, p<0.07 vs. sham; CAL+rAAV9-SelT: 0.46±0.02 ml/beat, p<0.05 vs. HF) and cardiac output (sham: 173±7; HF: 153±6, p<0.05 vs. sham; CAL+rAAV9-SelT: 178±7 ml/min, p<0.05 vs. HF) without change in heart rate or body weight. Conclusion Our results are the first to show that rAAV9-mediated-SelT gene therapy produces beneficial effects on cardiac remodeling and function in rats with HF. The author hereby declares no conflict of interest
0113 Epicardial progenitors are source of adipocyte in human atria Nadine Suffee * (1), Thomas Moore Moris (2), Gilles Dilanian (1), Patrick Farahmand (3), Catherine Rucker-Martin (4), Isabelle Dugail (1), Michel Pucéat (2), Stéphane Hatem (1) (1) Université Pierre et Marie Curie, INSERM U1166, Paris, France – (2) Faculté de Médecine, INSERM UMRS 910, Marseille, France – (3) APHP-GH Pitié-Salpêtrière, Institut du Coeur, Paris, France – (4) Centre Chirurgical Marie Lannelongue, Pediatric and Congenital Heart Diseases, Le Plessis-Robinson, France * Corresponding author: [email protected] (Nadine Suffee) Background and aims The accumulation of the adipose tissue (AT) in the sup- and subepicardium of the atrial myocardium is associated with a high risk of atrial fibrillation. Here we addressed the question of the cellular origin of atrial AT.
Methods Human right atrial specimen obtained during cardiac surgery were used for histological, biochemical studies and to harvest epicardial progenitors. Cells were characterized using flow cytometry, proteomic and genic expression assays and maintained in culture conditions, in some experiments they were co cultured with human atrial myocytes using a Transwell system. Results In atrial section, epicardial progenitors expressing the pre-adipocyte factor 1 (Pref-1) were detected in the epicardial and sub-epicardial layer which contained a number of progenitor cells referred as epicardic progenitorderived cells (EPDCs). Some EPDCs expressed Pref-1 suggesting that they could engaged in the adipocyte pathways. This was tested in vitro using EPDCs harvested from human right atrial samples (n=20). From 1st to 5-6 cells passages, EPDCs underwent an epithelial-to-mesenchymal transition (EMT), acquired mesenchymal phenotypes and could differentiate into osteocyte or chondrocyte. When cultured using an adipogenic medium, around 40±6% of EPDCs cells showed lipid droplet colored with oil red and expressed mature adipocyte perilipin marker. Culture medium enriched with the secretome of human atria too induced the differentiation of EPDCs in adipocytes (n=6). A shift from pre- to mature adipocytes was indicated by the detection of Pref-1 at 7 but not 30 days of culture, it was the opposite for perilipin. Human atrial myocyte could be the source of pro-adipogenic factor. In fact, after 7 days in coculture, EPDCs are differentiated into adipocytes (n=6). Conclusion Human EPDCs from atria can differentiate into adipocyte; factors secreted by atrial myocytes regulate this differentiation process. The author hereby declares no conflict of interest
0006 Proliferation of the cardiac precursor cells expressing the stem cell antigen-1 is controlled by activation of the natriuretic peptide receptors Stéphanie Rignault, Christelle Bielmann, Lucas Liaudet, Bernard Waeber, François Feihl, Nathalie Rosenblatt * CHU Vaudois, Lausanne, Suisse * Corresponding author: [email protected] (Nathalie Rosenblatt) A part of the cardioprotective role of the Brain Natriuretic Peptide (BNP) in mouse hearts is due to its effect on the cardiac precursor cell (CPC) proliferation and differentiation. Thus, in this study we identified the CPC subset able to respond to BNP as well as the signaling pathway involved. We demonstrated by immunohistochemistry and by flow cytometry analysis that the c-kit+ and the Sca-1+ cell subsets in neonatal and adult murine hearts express the NPR-A and NPR-B receptors and are thus able to be stimulated by BNP. In vitro, BNP only stimulated the proliferation of the Sca-1+ cells and not of the c-kit+ cells. Among Sca-1+ cells, BNP treatment led to increased number of Sca-1+ Nkx2.5+ cells, which were able to differentiate into cardiomyocytes. To determine by which receptor BNP acts on Sca-1+ cells to stimulate their proliferation, cells were isolated from neonatal hearts of mice deficient for the NPR-A (NPRA-KO) or NPR-B receptor. BNP stimulated the proliferation of the Sca-1+ NPR-A KO cells but not of the Sca-1+ cells lacking the NPR-B receptor, demonstrating that Sca-1+ cell proliferation is linked to NPR-B activation. This was confirmed by stimulating the Sca-1+ cells by the C-Natriuretic Peptide able also to activate the NPR-B receptor. BNP binding to NPR-B receptor led in Sca-1+ cells to Protein Kinase G activation and increased phosphorylation of phospholamban and p38. Reducing PKG activation inhibited BNP-induced-Sca-1+ cell proliferation, whereas reducing p38 phosphorylation increased Sca-1+ cell proliferation after BNP treatment. Phosphorylation of p38 was not mediated by BNP binding to NPR-B receptor but by its binding to NPR-A. In this work, we identified the Sca-1+ cells as being the targets of BNP in vitro and in vivo. BNP via NPR-B binding and PKG activation clearly stimulated the proliferation of the CPCs expressing Sca-1. Interestingly, is the dual role of the NPR-A and NPR-B receptors which control Sca-1+ cell proliferation. The author hereby declares no conflict of interest
© Elsevier Masson SAS. All rights reserved.
Archives of Cardiovascular Diseases Supplements (2016) 8, 255-257
Topic 28 – Cellular and gene therapy, stem cells April 08th, Friday 2016
0464 Selenoprotein T gene therapy with adeno-associated virus-9 improves cardiac function/remodeling in rats with heart failure Inès Boukhalfa * (1), Orianne Henri (1), Jean-Paul Henry (1), Youssef Anouar (2), Vincent Richard (1), Sahil Adriouch (3), Paul Mulder (1) (1) Université de Rouen, Rouen, France – (2) Université de Rouen, INSERM U982, Rouen, France – (3) Université de Rouen, INSERM U905, Rouen, France * Corresponding author: [email protected] (Inès Boukhalfa) Background Selenoprotein T (SelT) is a thioredoxin-like protein, which is abundantly but transiently expressed in the heart only during the embryonic development. Although SelT seems to play a limited role in adulthood, preliminary data from our laboratory showed that myocardial infarction increases SelT protein expression. This suggests that SelT may play a yet unrevealed role in cardiovascular diseases. Our studies have proven that SelT supplementation, using osmotic minipump delivery system, clearly improves cardiac remodeling and function, cardiac perfusion and decreases cardiac inflammation pathways after experimental heart failure (HF). Thus, we sought to investigate the cardiac effects of SelT over-expression by administration of recombinant adeno-associated virus 9 (rAAV9) in HF. Methods Wistar rats received a single intravenous injection of either rAAV9-SelT or saline serum one week after coronary artery ligation and left ventricular function/remodeling was determined 3 months after ligation by echocardiography. Results rAAV9-SelT (1.1011 virus-genome copies) resulted in an increased cardiac SelT expression (+150 %, p<0.05).This SelT over-expression reduced HF induced increase in left ventricular diastolic (sham: 6.14±0.16; HF: 10.83±0.36, p<0.05 vs. sham; HF+rAAV9-SelT: 9.92±0.21 mm, p<0.05 vs. HF) and systolic diameters (sham: 3.08±0.17; HF, 9.13±0.35, p<0.05 vs. sham; CAL+rAAV9-SelT: 8.14±0.27 mm, p<0.05 vs HF). Simultaneously, SelT improved stroke volume (sham: 0.49±0.02; HF: 0.41±0.01, p<0.07 vs. sham; CAL+rAAV9-SelT: 0.46±0.02 ml/beat, p<0.05 vs. HF) and cardiac output (sham: 173±7; HF: 153±6, p<0.05 vs. sham; CAL+rAAV9-SelT: 178±7 ml/min, p<0.05 vs. HF) without change in heart rate or body weight. Conclusion Our results are the first to show that rAAV9-mediated-SelT gene therapy produces beneficial effects on cardiac remodeling and function in rats with HF. The author hereby declares no conflict of interest
0113 Epicardial progenitors are source of adipocyte in human atria Nadine Suffee * (1), Thomas Moore Moris (2), Gilles Dilanian (1), Patrick Farahmand (3), Catherine Rucker-Martin (4), Isabelle Dugail (1), Michel Pucéat (2), Stéphane Hatem (1) (1) Université Pierre et Marie Curie, INSERM U1166, Paris, France – (2) Faculté de Médecine, INSERM UMRS 910, Marseille, France – (3) APHP-GH Pitié-Salpêtrière, Institut du Coeur, Paris, France – (4) Centre Chirurgical Marie Lannelongue, Pediatric and Congenital Heart Diseases, Le Plessis-Robinson, France * Corresponding author: [email protected] (Nadine Suffee) Background and aims The accumulation of the adipose tissue (AT) in the sup- and subepicardium of the atrial myocardium is associated with a high risk of atrial fibrillation. Here we addressed the question of the cellular origin of atrial AT.
Methods Human right atrial specimen obtained during cardiac surgery were used for histological, biochemical studies and to harvest epicardial progenitors. Cells were characterized using flow cytometry, proteomic and genic expression assays and maintained in culture conditions, in some experiments they were co cultured with human atrial myocytes using a Transwell system. Results In atrial section, epicardial progenitors expressing the pre-adipocyte factor 1 (Pref-1) were detected in the epicardial and sub-epicardial layer which contained a number of progenitor cells referred as epicardic progenitorderived cells (EPDCs). Some EPDCs expressed Pref-1 suggesting that they could engaged in the adipocyte pathways. This was tested in vitro using EPDCs harvested from human right atrial samples (n=20). From 1st to 5-6 cells passages, EPDCs underwent an epithelial-to-mesenchymal transition (EMT), acquired mesenchymal phenotypes and could differentiate into osteocyte or chondrocyte. When cultured using an adipogenic medium, around 40±6% of EPDCs cells showed lipid droplet colored with oil red and expressed mature adipocyte perilipin marker. Culture medium enriched with the secretome of human atria too induced the differentiation of EPDCs in adipocytes (n=6). A shift from pre- to mature adipocytes was indicated by the detection of Pref-1 at 7 but not 30 days of culture, it was the opposite for perilipin. Human atrial myocyte could be the source of pro-adipogenic factor. In fact, after 7 days in coculture, EPDCs are differentiated into adipocytes (n=6). Conclusion Human EPDCs from atria can differentiate into adipocyte; factors secreted by atrial myocytes regulate this differentiation process. The author hereby declares no conflict of interest
0006 Proliferation of the cardiac precursor cells expressing the stem cell antigen-1 is controlled by activation of the natriuretic peptide receptors Stéphanie Rignault, Christelle Bielmann, Lucas Liaudet, Bernard Waeber, François Feihl, Nathalie Rosenblatt * CHU Vaudois, Lausanne, Suisse * Corresponding author: [email protected] (Nathalie Rosenblatt) A part of the cardioprotective role of the Brain Natriuretic Peptide (BNP) in mouse hearts is due to its effect on the cardiac precursor cell (CPC) proliferation and differentiation. Thus, in this study we identified the CPC subset able to respond to BNP as well as the signaling pathway involved. We demonstrated by immunohistochemistry and by flow cytometry analysis that the c-kit+ and the Sca-1+ cell subsets in neonatal and adult murine hearts express the NPR-A and NPR-B receptors and are thus able to be stimulated by BNP. In vitro, BNP only stimulated the proliferation of the Sca-1+ cells and not of the c-kit+ cells. Among Sca-1+ cells, BNP treatment led to increased number of Sca-1+ Nkx2.5+ cells, which were able to differentiate into cardiomyocytes. To determine by which receptor BNP acts on Sca-1+ cells to stimulate their proliferation, cells were isolated from neonatal hearts of mice deficient for the NPR-A (NPRA-KO) or NPR-B receptor. BNP stimulated the proliferation of the Sca-1+ NPR-A KO cells but not of the Sca-1+ cells lacking the NPR-B receptor, demonstrating that Sca-1+ cell proliferation is linked to NPR-B activation. This was confirmed by stimulating the Sca-1+ cells by the C-Natriuretic Peptide able also to activate the NPR-B receptor. BNP binding to NPR-B receptor led in Sca-1+ cells to Protein Kinase G activation and increased phosphorylation of phospholamban and p38. Reducing PKG activation inhibited BNP-induced-Sca-1+ cell proliferation, whereas reducing p38 phosphorylation increased Sca-1+ cell proliferation after BNP treatment. Phosphorylation of p38 was not mediated by BNP binding to NPR-B receptor but by its binding to NPR-A. In this work, we identified the Sca-1+ cells as being the targets of BNP in vitro and in vivo. BNP via NPR-B binding and PKG activation clearly stimulated the proliferation of the CPCs expressing Sca-1. Interestingly, is the dual role of the NPR-A and NPR-B receptors which control Sca-1+ cell proliferation. The author hereby declares no conflict of interest
© Elsevier Masson SAS. All rights reserved.