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Strategy for Tissue-Engineering Vasculature with Biodegradable Scaffold in Congenital Heart Diseases
S130 Journal of Cardiac Failure Vol. 17 No. 9S September 2011
Symposium 5 S5-1 Cardiac Regeneration by Intrinsic Cardiac Stem Cells TOSHIO NAGAI1, MASATO KANDA1, MEILAN LIU1, NAOMICHI KONDO1, TOSHINAO TAKAHASHI1, KATSUHISA MATSUURA2, YOSHIO KOBAYASHI1, ISSEI KOMURO3 1 Cardiovascular Science and medicine, Chiba University Graduate School of Medicine, 2Department of Cardiology, Tokyo Women’s Medical University, Tokyo Japan, 3Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Osaka Japan Cell therapy has been expected to be a new therapy for severe heart failure. We have reported that transplantation of cardiac progenitor cells with self-assembling nanopeptides prevents cardiac remodeling and dysfunction through angiogenesis, inhibition of apoptosis and myocardial regeneration. The secreted molecules specific to cardiac progenitors, such as VCAM-1, play a crucial role in the beneficial effects. Recently we identified another secreted molecules, which attenuated the inflammation through inhibiting transendothelial migration of leukocytes. Besides the transplanted cardiac progenitors, the regenerative capacity of intrinsic cardiac progenitors has not been elucidated. The lineage mapping analysis by using a double-transgenic alphaMHC-MerCreMer/CAG-LacZ mouse enabled us to identify newly formed cardiomyocytes after treatment with 4-OH-tamoxifen. We identified that leukemia inhibitory factor promoted cardiomyogenesis after myocardial infarction. Although it is still unclear whether proliferation, anti-apoptosis, or differentiation of cardiac stem cells is related to the underlying mechanism, however, this finding suggests that the intrinsic cardiac stem cell is a potent target for cardiac regeneration therapy.
S5-2 First-in-man Cell Therapy Clinical Trial for Heart Failure -AutoLogous human Cardic-Derived Stem Cell to Treat Ischemic cardiomyopathy (ALCADIA) HIROAKI MATSUBARA Department of Cradiovascular Medicine, Kyoto Frefectural University School of Medicine, Kyoto, Japan Although human cardiac stem cell transplantation had functional benefits in the recovery in experimental myocardial infarction, the major barrier limiting its clinical application is the death of the most of the transplanted cells and poor cardiac differentiation in the host environment. Using the identical technique as clonally cell isolation from experimental animals, we generated human cardiosphere-derived cell (hCDC) enriched Es-marker genes with mesenchymal features, which were obtained from cardiac endomyocardial biopsy samples. bFGF possesses properties to promote stem cell proliferation, and formation of sufficient microvascular network created by bFGF is critical for long-term survival of transplanted donor cells. To investigate the effect of hCDC transplantation with controlled-release bFGF using biodegradable gelatin, we performed preclinical trials of chronically instrumented pigs. When combined with bFGF, hCDC transplantation specifically improved cardiac function of experimental pigs accompanied with enhancing hCDC engraftment, contributing to effective cardiovascular regeneration. Our results demonstrate a promising approach for achieving a remarkably functional cardiac repair by a combination of hCDC transplantation with bFGF-incorporating hydrogel. We have already started Phase I Clinical trial termed by ARCADIA for patients with severe heart failure combined with bypass surgery. The result from 4 patients will be lectured in the symposium.
S5-3 ES & iPS Cell Research for Cardiovascular Regeneration JUN YAMASHITA Institute for Frontier Medical Sciences, Kyoto University We have been investigating cardiovascular cell differentiation and regeneration using embryonic stem (ES) and induced pluripotent stem (iPS) cells. Previously, we established an ES cell differentiation system for cardiovascular cells using Flk1+ cells as common progenitors (Nature, 2000). We recently found an efficient expansion method for cardimyocytes and cardiac progenitors from ES cells. That is, an immunosuppressant, cyclosporin-A (CSA), showed a novel effect specifically acting on mesoderm cells to drastically increase cardiac progenitors as well as cardiomyocytes
(Biochem Biophys Res Commun, 2009). Recently, novel pluripotent stem cell lines, iPS cells, were established from mouse and human somatic cells. We applied our ES cell system to mouse iPS cells, and succeeded in systematically inducing cardiovascular cells from iPS cells (Circulation, 2008). Combining our technologies in ES cells and iPS cells, we recently revealed that cardiac cells were efficiently induced from mouse and human iPS cells by CSA treatment (PLoS One, 2011). Now we are exploring cell transplantation methods using the cell sheets technology (Shimizu, Curr Pharm Des, 2009) and small molecules for cardiac regeneration. These studies would largely contribute to cardiovascular regeneration with iPS cells.
S5-4 Direct Reprogramming of Fibroblasts into Cardiomyocytes-A New Approach for Cardiac Regeneration MASAKI IEDA Department of Clinical and Molecular Cardiovascular Research, Keio University School of Medicine, Tokyo, Japan Generating iPS cells by a combination of defined factors demonstrated that somatic cells can be induced to change their cell fate. Many fibroblasts exist in the post-natal heart, but no single master regulator of cardiac reprogramming has been identified. We postulated that not a single, but a combination of cardiogenic factors may reprogram fibroblasts into cardiomyocytes. We selected 14 candidate factors which are specifically expressed in embryonic cardiomyocytes and exhibited severe developmental cardiac defects and embryonic lethality when mutated. Among them, a combination of three developmental transcription factors rapidly and efficiently reprogrammed post-natal cardiac fibroblasts directly into cardiomyocytes. Induced cardiomyocytes expressed cardiac-specific proteins, exhibited a global gene expression profile and an epigenetic state similar to cardiomyocytes, and contracted spontaneously. Induced cardiomyocytes were not converted into the cardiac progenitor state for reprogramming, but rather they were directly reprogrammed into differentiated cardiomyocytes by defined factors. Moreover, fibroblast cells transplanted into mouse hearts one day after transduction of the three factors differentiated into cardiomyocytes in vivo. These findings demonstrate that functional cardiomyocytes can be directly reprogrammed from differentiated somatic cells by defined factors. Reprogramming the vast pool of endogenous cardiac fibroblasts might provide a source of cardiomyocytes for regenerative therapies.
S5-5 Strategy for Tissue-Engineering Vasculature with Biodegradable Scaffold in Congenital Heart Diseases GOKI MATSUMURA1, KENJI YAMAZAKI1, NAOTAKA NITTA2, YUKI SAKAMOTO3, SHOJIRO MATSUDA3 1 Cardiovascular Surgery, Tokyo Women’s Medical University, Tokyo, Japan, 2 Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan, 3GunzeLtd, Research and Developing Center, Kyoto, Japan The use of foreign materials is necessary to repair complex heart defects. However, long-term studies of the efficacy of these materials have revealed several materialrelated failures, such as stenosis, thromboembolization, and calcium deposition. To solve these problems and to improve the treatment of children who require implantation of biocompatible materials possessing growth potential, we have been pursuing the development of optimal materials. We previously reported the advantages of using biodegradable scaffolds seeded with autologous cells as tissue-engineered vasculature (TEV) in canine models and in a human clinical study. Since May 1999, we have performed cardiac surgery using tissue-engineered materials, and the number of patients has increased up to 47. There was no graft-related mortality and no evidence of aneurysm formation, graft rupture, graft infection, or ectopic calcification in the late-term results. However, some patients had graft stenosis, which required percutaneous angioplasty. We then explored a new scaffold without cell seeding to overcome the graft stenosis of TEV and complicated protocol, and achieved acceptable long-term results in an animal model. This scaffold can be implanted by a simple cost-effective procedure, because no cell preparation and seeding are necessary. This novel strategy can easily be applied in near future to the treatment of patients who require surgical interventions with artificial grafts to provide a better quality of life.
Symposium 5 S5-1 Cardiac Regeneration by Intrinsic Cardiac Stem Cells TOSHIO NAGAI1, MASATO KANDA1, MEILAN LIU1, NAOMICHI KONDO1, TOSHINAO TAKAHASHI1, KATSUHISA MATSUURA2, YOSHIO KOBAYASHI1, ISSEI KOMURO3 1 Cardiovascular Science and medicine, Chiba University Graduate School of Medicine, 2Department of Cardiology, Tokyo Women’s Medical University, Tokyo Japan, 3Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Osaka Japan Cell therapy has been expected to be a new therapy for severe heart failure. We have reported that transplantation of cardiac progenitor cells with self-assembling nanopeptides prevents cardiac remodeling and dysfunction through angiogenesis, inhibition of apoptosis and myocardial regeneration. The secreted molecules specific to cardiac progenitors, such as VCAM-1, play a crucial role in the beneficial effects. Recently we identified another secreted molecules, which attenuated the inflammation through inhibiting transendothelial migration of leukocytes. Besides the transplanted cardiac progenitors, the regenerative capacity of intrinsic cardiac progenitors has not been elucidated. The lineage mapping analysis by using a double-transgenic alphaMHC-MerCreMer/CAG-LacZ mouse enabled us to identify newly formed cardiomyocytes after treatment with 4-OH-tamoxifen. We identified that leukemia inhibitory factor promoted cardiomyogenesis after myocardial infarction. Although it is still unclear whether proliferation, anti-apoptosis, or differentiation of cardiac stem cells is related to the underlying mechanism, however, this finding suggests that the intrinsic cardiac stem cell is a potent target for cardiac regeneration therapy.
S5-2 First-in-man Cell Therapy Clinical Trial for Heart Failure -AutoLogous human Cardic-Derived Stem Cell to Treat Ischemic cardiomyopathy (ALCADIA) HIROAKI MATSUBARA Department of Cradiovascular Medicine, Kyoto Frefectural University School of Medicine, Kyoto, Japan Although human cardiac stem cell transplantation had functional benefits in the recovery in experimental myocardial infarction, the major barrier limiting its clinical application is the death of the most of the transplanted cells and poor cardiac differentiation in the host environment. Using the identical technique as clonally cell isolation from experimental animals, we generated human cardiosphere-derived cell (hCDC) enriched Es-marker genes with mesenchymal features, which were obtained from cardiac endomyocardial biopsy samples. bFGF possesses properties to promote stem cell proliferation, and formation of sufficient microvascular network created by bFGF is critical for long-term survival of transplanted donor cells. To investigate the effect of hCDC transplantation with controlled-release bFGF using biodegradable gelatin, we performed preclinical trials of chronically instrumented pigs. When combined with bFGF, hCDC transplantation specifically improved cardiac function of experimental pigs accompanied with enhancing hCDC engraftment, contributing to effective cardiovascular regeneration. Our results demonstrate a promising approach for achieving a remarkably functional cardiac repair by a combination of hCDC transplantation with bFGF-incorporating hydrogel. We have already started Phase I Clinical trial termed by ARCADIA for patients with severe heart failure combined with bypass surgery. The result from 4 patients will be lectured in the symposium.
S5-3 ES & iPS Cell Research for Cardiovascular Regeneration JUN YAMASHITA Institute for Frontier Medical Sciences, Kyoto University We have been investigating cardiovascular cell differentiation and regeneration using embryonic stem (ES) and induced pluripotent stem (iPS) cells. Previously, we established an ES cell differentiation system for cardiovascular cells using Flk1+ cells as common progenitors (Nature, 2000). We recently found an efficient expansion method for cardimyocytes and cardiac progenitors from ES cells. That is, an immunosuppressant, cyclosporin-A (CSA), showed a novel effect specifically acting on mesoderm cells to drastically increase cardiac progenitors as well as cardiomyocytes
(Biochem Biophys Res Commun, 2009). Recently, novel pluripotent stem cell lines, iPS cells, were established from mouse and human somatic cells. We applied our ES cell system to mouse iPS cells, and succeeded in systematically inducing cardiovascular cells from iPS cells (Circulation, 2008). Combining our technologies in ES cells and iPS cells, we recently revealed that cardiac cells were efficiently induced from mouse and human iPS cells by CSA treatment (PLoS One, 2011). Now we are exploring cell transplantation methods using the cell sheets technology (Shimizu, Curr Pharm Des, 2009) and small molecules for cardiac regeneration. These studies would largely contribute to cardiovascular regeneration with iPS cells.
S5-4 Direct Reprogramming of Fibroblasts into Cardiomyocytes-A New Approach for Cardiac Regeneration MASAKI IEDA Department of Clinical and Molecular Cardiovascular Research, Keio University School of Medicine, Tokyo, Japan Generating iPS cells by a combination of defined factors demonstrated that somatic cells can be induced to change their cell fate. Many fibroblasts exist in the post-natal heart, but no single master regulator of cardiac reprogramming has been identified. We postulated that not a single, but a combination of cardiogenic factors may reprogram fibroblasts into cardiomyocytes. We selected 14 candidate factors which are specifically expressed in embryonic cardiomyocytes and exhibited severe developmental cardiac defects and embryonic lethality when mutated. Among them, a combination of three developmental transcription factors rapidly and efficiently reprogrammed post-natal cardiac fibroblasts directly into cardiomyocytes. Induced cardiomyocytes expressed cardiac-specific proteins, exhibited a global gene expression profile and an epigenetic state similar to cardiomyocytes, and contracted spontaneously. Induced cardiomyocytes were not converted into the cardiac progenitor state for reprogramming, but rather they were directly reprogrammed into differentiated cardiomyocytes by defined factors. Moreover, fibroblast cells transplanted into mouse hearts one day after transduction of the three factors differentiated into cardiomyocytes in vivo. These findings demonstrate that functional cardiomyocytes can be directly reprogrammed from differentiated somatic cells by defined factors. Reprogramming the vast pool of endogenous cardiac fibroblasts might provide a source of cardiomyocytes for regenerative therapies.
S5-5 Strategy for Tissue-Engineering Vasculature with Biodegradable Scaffold in Congenital Heart Diseases GOKI MATSUMURA1, KENJI YAMAZAKI1, NAOTAKA NITTA2, YUKI SAKAMOTO3, SHOJIRO MATSUDA3 1 Cardiovascular Surgery, Tokyo Women’s Medical University, Tokyo, Japan, 2 Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan, 3GunzeLtd, Research and Developing Center, Kyoto, Japan The use of foreign materials is necessary to repair complex heart defects. However, long-term studies of the efficacy of these materials have revealed several materialrelated failures, such as stenosis, thromboembolization, and calcium deposition. To solve these problems and to improve the treatment of children who require implantation of biocompatible materials possessing growth potential, we have been pursuing the development of optimal materials. We previously reported the advantages of using biodegradable scaffolds seeded with autologous cells as tissue-engineered vasculature (TEV) in canine models and in a human clinical study. Since May 1999, we have performed cardiac surgery using tissue-engineered materials, and the number of patients has increased up to 47. There was no graft-related mortality and no evidence of aneurysm formation, graft rupture, graft infection, or ectopic calcification in the late-term results. However, some patients had graft stenosis, which required percutaneous angioplasty. We then explored a new scaffold without cell seeding to overcome the graft stenosis of TEV and complicated protocol, and achieved acceptable long-term results in an animal model. This scaffold can be implanted by a simple cost-effective procedure, because no cell preparation and seeding are necessary. This novel strategy can easily be applied in near future to the treatment of patients who require surgical interventions with artificial grafts to provide a better quality of life.