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Novel Purification Method of Pluripotent Stem Cell-derived Cardiomyocytes Using Metabolome Analysis
The 16th Annual Scientific Meeting
JHFS
S145
Symposium 14 S14-1
S14-4
Novel Purification Method of Pluripotent Stem Cell-derived Cardiomyocytes Using Metabolome Analysis SHUGO TOHYAMA, FUMIYUKI HATTORI, MOTOAKI SANO, KEIICHI FUKUDA Department of Cardiology, Keio University School of Medicine, Tokyo, Japan
From Bench to Bedside Work of Myoblast Sheet-based Myocardial Regeneration Therapy SHIGERU MIYAGAWA, KOUICHI TODA, HIROYUKI NISHI, YASUSHI YOSHIKAWA, SATSUKI FUKUSHIMA, DAISUKE YOSHIOKA, SOUKICHI KAMATA, TETSUYA SAITO, ATSUHIRO SAITO, YOSHIKI SAWA Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Japan
Mass production of highly purified cardiomyocytes is a critical bottleneck in realizing heart regenerative therapy. Our recently established non-genetic “mitochondrial method” can highly purify cardiomyocytes, but remains insufficient for clinical-scale purification. Here we report a novel method to purify bulk cardiomyocytes from pluripotent stem cell (PSC)-derivatives based on the findings from transcriptome and metabolome analyses, which highlighted marked biochemical differences in glucose and lactate metabolism between embryonic stem cells (ESCs) and cardiomyocytes. We cultured ESC derivatives with glucose-depleted culture medium containing abundant lactate, in which only cardiomyocytes eventually survived. We obtained cardiomyocytes of O 98% purity that did not form teratomas after transplantation. Furthermore, metabolome analysis using [13C]-labeled lactate revealed that cardiomyocytes use lactate for both ATP production and biomass synthesis for survival. We believe that our novel technology will resolve the bottleneck and directly facilitate human heart-regenerative therapies.
S14-2 Cell Sheet-based Cardiac Tissue Engineering for Regenerative Medicine KATSUHISA MATSUURA1, TATSUYA SHIMIZU1, NOBUHISA HAGIWARA2, TERUO OKANO1 1 Institute of Advanced Biomedical Engineering and Science (TWIns), Tokyo Women’s Medical University, Tokyo, Japan, 2Department of Cardiology, Tokyo Women’s Medical University, Tokyo, Japan The bioengineered functional heart tissue is thought to be promising for not only the regenerative medicine, but also the heart tissue models. We have established so called “cell sheet engineering” to create functional tissue sheets for treating a wide range of diseases such as corneal dysfunction, esophageal cancer and heart failure. Now we are attempting to build automated fabrication systems to produce large quantities of cell sheets and thick tissues by combining biomedical and engineering technologies. On the other hand, current cardiac cell therapy repairs the injured heart via mainly paracrine mechanisms, thus the fabrication of thickened cardiac tissue that can directly contribute to the cardiac function leads to the next generation of cardiac regenerative medicine. Recently we have developed the methods to create layered and vascularized tissues for organ-like systems such as the heart. Furthermore the development of three-dimensional bioreactorbased culture system of pluripotent stem cells enables us to collect a large amount of cardiovascular cells and create the human cardiac tissue. Cell sheet engineering, therefore, might provide a novel alternative approach for tissue based regenerative medicine.
S14-3 Cardiac Progenitor Cell Infusion in Patients with Hypoplastic Left Heart Syndrome: a Prospective Phase 1 Clinical Trial HIDEMASA OH1, SUGURU TARUI2, SHINICHI OTSUKI3, SHUNJI SANO2 1 Center for Innovative Clinical Medicine, Okayama University Hospital, Okayama, Japan, 2Department of Cardiac Surgery, Okayama University Hospital, Okayama, Japan, 3Department of Pediatric Cardiology, Okayama University Hospital, Okayama, Japan Hypoplastic left heart syndrome (HLHS) is characterized by an obstruction of left ventricle in association with underdevelopment of the left ventricular chamber morphogenesis that is not capable of supporting systemic cardiac output. Endogenous cardiac progenitor cells (CPCs) from adult mammalian heart were discovered a decade ago and the following phase 1 trials have recently suggested that transplantation of CPCs is feasible and safe in patients after recent infarction. Intensive investigations have uncovered the mechanistic potentials that mammalian hearts may have a capacity to self-renew the cardiac muscles after acute insult through de-differentiation of preexisting cardiomyocytes and/or trans-differentiation of a population of resident progenitors. This process is apparently demanded on the age indicated a possibility that human cardiomyocyte replenishment may be efficiently occurred during the early development in postnatal period. As a preclinical study, we report here that CPC infusion in rat model of right ventricular pressure overload significantly reduced the fibrotic area that was endorsed by a successful engraftment and cardiomyocytes differentiation in the transplanted hearts. These results gave us a rationale to conduct a phase 1 clinical trial to treat infants with HLHS after staged palliative-reconstructions. We show here that transcoronary infusion of autologous CPCs is feasible and safe, warranting phase 2 trials to verify the efficacy in patients with HLHS toward the development of therapeutic strategies.
LVAD implantation and Heart transplantation have been well accepted as the ultimate lifesaving means of supporting end-staged heart failure patients. However, due to the limited durability of the LVAD and the shortage of donors, there are some limitations in these procedures. In this clinical situation, we developed cell sheet technology experimentally and introduced this to the treatment of severely damaged myocardium in clinical setting as translational research. In a series of pre-clinical experiments using animal heart failure model, we proved that myoblast sheets could heal the impaired heart mainly by cytokine paracrine effect. Evidenced by these pre-clinical experiments, we applied myoblast sheets to 4 DCM patient receiving LVAD and 2 patients showed the recovery from LVAD. And we implanted 12 patients (ICM 7, DCM 5) without LVAD and some patients showed LV reverse remodeling. To improve the effectiveness of myoblast sheet, we implanted a large number of cell sheets combined with omentum implantation. Omentum implantation could activate the paracrine effect of myoblast sheets and a large number of implanted cells survived on the epicardium with rich vascular network, leading to the enhancement of myocardial regeneration in porcine MI model. Cell sheet technology may be a promising armamentarium for healing severely damaged myocardium.
JHFS
S145
Symposium 14 S14-1
S14-4
Novel Purification Method of Pluripotent Stem Cell-derived Cardiomyocytes Using Metabolome Analysis SHUGO TOHYAMA, FUMIYUKI HATTORI, MOTOAKI SANO, KEIICHI FUKUDA Department of Cardiology, Keio University School of Medicine, Tokyo, Japan
From Bench to Bedside Work of Myoblast Sheet-based Myocardial Regeneration Therapy SHIGERU MIYAGAWA, KOUICHI TODA, HIROYUKI NISHI, YASUSHI YOSHIKAWA, SATSUKI FUKUSHIMA, DAISUKE YOSHIOKA, SOUKICHI KAMATA, TETSUYA SAITO, ATSUHIRO SAITO, YOSHIKI SAWA Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Japan
Mass production of highly purified cardiomyocytes is a critical bottleneck in realizing heart regenerative therapy. Our recently established non-genetic “mitochondrial method” can highly purify cardiomyocytes, but remains insufficient for clinical-scale purification. Here we report a novel method to purify bulk cardiomyocytes from pluripotent stem cell (PSC)-derivatives based on the findings from transcriptome and metabolome analyses, which highlighted marked biochemical differences in glucose and lactate metabolism between embryonic stem cells (ESCs) and cardiomyocytes. We cultured ESC derivatives with glucose-depleted culture medium containing abundant lactate, in which only cardiomyocytes eventually survived. We obtained cardiomyocytes of O 98% purity that did not form teratomas after transplantation. Furthermore, metabolome analysis using [13C]-labeled lactate revealed that cardiomyocytes use lactate for both ATP production and biomass synthesis for survival. We believe that our novel technology will resolve the bottleneck and directly facilitate human heart-regenerative therapies.
S14-2 Cell Sheet-based Cardiac Tissue Engineering for Regenerative Medicine KATSUHISA MATSUURA1, TATSUYA SHIMIZU1, NOBUHISA HAGIWARA2, TERUO OKANO1 1 Institute of Advanced Biomedical Engineering and Science (TWIns), Tokyo Women’s Medical University, Tokyo, Japan, 2Department of Cardiology, Tokyo Women’s Medical University, Tokyo, Japan The bioengineered functional heart tissue is thought to be promising for not only the regenerative medicine, but also the heart tissue models. We have established so called “cell sheet engineering” to create functional tissue sheets for treating a wide range of diseases such as corneal dysfunction, esophageal cancer and heart failure. Now we are attempting to build automated fabrication systems to produce large quantities of cell sheets and thick tissues by combining biomedical and engineering technologies. On the other hand, current cardiac cell therapy repairs the injured heart via mainly paracrine mechanisms, thus the fabrication of thickened cardiac tissue that can directly contribute to the cardiac function leads to the next generation of cardiac regenerative medicine. Recently we have developed the methods to create layered and vascularized tissues for organ-like systems such as the heart. Furthermore the development of three-dimensional bioreactorbased culture system of pluripotent stem cells enables us to collect a large amount of cardiovascular cells and create the human cardiac tissue. Cell sheet engineering, therefore, might provide a novel alternative approach for tissue based regenerative medicine.
S14-3 Cardiac Progenitor Cell Infusion in Patients with Hypoplastic Left Heart Syndrome: a Prospective Phase 1 Clinical Trial HIDEMASA OH1, SUGURU TARUI2, SHINICHI OTSUKI3, SHUNJI SANO2 1 Center for Innovative Clinical Medicine, Okayama University Hospital, Okayama, Japan, 2Department of Cardiac Surgery, Okayama University Hospital, Okayama, Japan, 3Department of Pediatric Cardiology, Okayama University Hospital, Okayama, Japan Hypoplastic left heart syndrome (HLHS) is characterized by an obstruction of left ventricle in association with underdevelopment of the left ventricular chamber morphogenesis that is not capable of supporting systemic cardiac output. Endogenous cardiac progenitor cells (CPCs) from adult mammalian heart were discovered a decade ago and the following phase 1 trials have recently suggested that transplantation of CPCs is feasible and safe in patients after recent infarction. Intensive investigations have uncovered the mechanistic potentials that mammalian hearts may have a capacity to self-renew the cardiac muscles after acute insult through de-differentiation of preexisting cardiomyocytes and/or trans-differentiation of a population of resident progenitors. This process is apparently demanded on the age indicated a possibility that human cardiomyocyte replenishment may be efficiently occurred during the early development in postnatal period. As a preclinical study, we report here that CPC infusion in rat model of right ventricular pressure overload significantly reduced the fibrotic area that was endorsed by a successful engraftment and cardiomyocytes differentiation in the transplanted hearts. These results gave us a rationale to conduct a phase 1 clinical trial to treat infants with HLHS after staged palliative-reconstructions. We show here that transcoronary infusion of autologous CPCs is feasible and safe, warranting phase 2 trials to verify the efficacy in patients with HLHS toward the development of therapeutic strategies.
LVAD implantation and Heart transplantation have been well accepted as the ultimate lifesaving means of supporting end-staged heart failure patients. However, due to the limited durability of the LVAD and the shortage of donors, there are some limitations in these procedures. In this clinical situation, we developed cell sheet technology experimentally and introduced this to the treatment of severely damaged myocardium in clinical setting as translational research. In a series of pre-clinical experiments using animal heart failure model, we proved that myoblast sheets could heal the impaired heart mainly by cytokine paracrine effect. Evidenced by these pre-clinical experiments, we applied myoblast sheets to 4 DCM patient receiving LVAD and 2 patients showed the recovery from LVAD. And we implanted 12 patients (ICM 7, DCM 5) without LVAD and some patients showed LV reverse remodeling. To improve the effectiveness of myoblast sheet, we implanted a large number of cell sheets combined with omentum implantation. Omentum implantation could activate the paracrine effect of myoblast sheets and a large number of implanted cells survived on the epicardium with rich vascular network, leading to the enhancement of myocardial regeneration in porcine MI model. Cell sheet technology may be a promising armamentarium for healing severely damaged myocardium.