Direct reprogramming of fibroblasts into endothelial cells capable of angiogenesis and reendothelialization in tissue-engineered vessels

The generation of induced pluripotent stem (iPS) cells is an important tool for regenerative medicine. However, the main restriction is the risk of tumor development. In this study we found that during the early stages of somatic cell reprogramming toward a pluripotent state, specific gene expressio...

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Published in:	Proceedings of the National Academy of Sciences - PNAS Vol. 109; no. 34; p. 13793
Main Authors:	Margariti, Andriana, Winkler, Bernhard, Karamariti, Eirini, Zampetaki, Anna, Tsai, Tsung-neng, Baban, Dilair, Ragoussis, Jiannis, Huang, Yi, Han, Jing-Dong J, Zeng, Lingfang, Hu, Yanhua, Xu, Qingbo
Format:	Journal Article
Language:	English
Published:	United States 21.08.2012
Subjects:	Animals Antigens, CD - genetics Aorta - pathology Cadherins - genetics Cell Differentiation Cells, Cultured Cellular Reprogramming Endothelial Cells - cytology Fibroblasts - cytology Fibroblasts - metabolism Humans Induced Pluripotent Stem Cells - cytology Kruppel-Like Factor 4 Mice Mice, SCID Models, Genetic Neovascularization, Pathologic Promoter Regions, Genetic Stem Cells - cytology Stress, Mechanical Tissue Engineering - methods
ISSN:	1091-6490, 1091-6490
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Abstract	The generation of induced pluripotent stem (iPS) cells is an important tool for regenerative medicine. However, the main restriction is the risk of tumor development. In this study we found that during the early stages of somatic cell reprogramming toward a pluripotent state, specific gene expression patterns are altered. Therefore, we developed a method to generate partial-iPS (PiPS) cells by transferring four reprogramming factors (OCT4, SOX2, KLF4, and c-MYC) to human fibroblasts for 4 d. PiPS cells did not form tumors in vivo and clearly displayed the potential to differentiate into endothelial cells (ECs) in response to defined media and culture conditions. To clarify the mechanism of PiPS cell differentiation into ECs, SET translocation (myeloid leukemia-associated) (SET) similar protein (SETSIP) was indentified to be induced during somatic cell reprogramming. Importantly, when PiPS cells were treated with VEGF, SETSIP was translocated to the cell nucleus, directly bound to the VE-cadherin promoter, increasing vascular endothelial-cadherin (VE-cadherin) expression levels and EC differentiation. Functionally, PiPS-ECs improved neovascularization and blood flow recovery in a hindlimb ischemic model. Furthermore, PiPS-ECs displayed good attachment, stabilization, patency, and typical vascular structure when seeded on decellularized vessel scaffolds. These findings indicate that reprogramming of fibroblasts into ECs via SETSIP and VEGF has a potential clinical application.
AbstractList	The generation of induced pluripotent stem (iPS) cells is an important tool for regenerative medicine. However, the main restriction is the risk of tumor development. In this study we found that during the early stages of somatic cell reprogramming toward a pluripotent state, specific gene expression patterns are altered. Therefore, we developed a method to generate partial-iPS (PiPS) cells by transferring four reprogramming factors (OCT4, SOX2, KLF4, and c-MYC) to human fibroblasts for 4 d. PiPS cells did not form tumors in vivo and clearly displayed the potential to differentiate into endothelial cells (ECs) in response to defined media and culture conditions. To clarify the mechanism of PiPS cell differentiation into ECs, SET translocation (myeloid leukemia-associated) (SET) similar protein (SETSIP) was indentified to be induced during somatic cell reprogramming. Importantly, when PiPS cells were treated with VEGF, SETSIP was translocated to the cell nucleus, directly bound to the VE-cadherin promoter, increasing vascular endothelial-cadherin (VE-cadherin) expression levels and EC differentiation. Functionally, PiPS-ECs improved neovascularization and blood flow recovery in a hindlimb ischemic model. Furthermore, PiPS-ECs displayed good attachment, stabilization, patency, and typical vascular structure when seeded on decellularized vessel scaffolds. These findings indicate that reprogramming of fibroblasts into ECs via SETSIP and VEGF has a potential clinical application.The generation of induced pluripotent stem (iPS) cells is an important tool for regenerative medicine. However, the main restriction is the risk of tumor development. In this study we found that during the early stages of somatic cell reprogramming toward a pluripotent state, specific gene expression patterns are altered. Therefore, we developed a method to generate partial-iPS (PiPS) cells by transferring four reprogramming factors (OCT4, SOX2, KLF4, and c-MYC) to human fibroblasts for 4 d. PiPS cells did not form tumors in vivo and clearly displayed the potential to differentiate into endothelial cells (ECs) in response to defined media and culture conditions. To clarify the mechanism of PiPS cell differentiation into ECs, SET translocation (myeloid leukemia-associated) (SET) similar protein (SETSIP) was indentified to be induced during somatic cell reprogramming. Importantly, when PiPS cells were treated with VEGF, SETSIP was translocated to the cell nucleus, directly bound to the VE-cadherin promoter, increasing vascular endothelial-cadherin (VE-cadherin) expression levels and EC differentiation. Functionally, PiPS-ECs improved neovascularization and blood flow recovery in a hindlimb ischemic model. Furthermore, PiPS-ECs displayed good attachment, stabilization, patency, and typical vascular structure when seeded on decellularized vessel scaffolds. These findings indicate that reprogramming of fibroblasts into ECs via SETSIP and VEGF has a potential clinical application. The generation of induced pluripotent stem (iPS) cells is an important tool for regenerative medicine. However, the main restriction is the risk of tumor development. In this study we found that during the early stages of somatic cell reprogramming toward a pluripotent state, specific gene expression patterns are altered. Therefore, we developed a method to generate partial-iPS (PiPS) cells by transferring four reprogramming factors (OCT4, SOX2, KLF4, and c-MYC) to human fibroblasts for 4 d. PiPS cells did not form tumors in vivo and clearly displayed the potential to differentiate into endothelial cells (ECs) in response to defined media and culture conditions. To clarify the mechanism of PiPS cell differentiation into ECs, SET translocation (myeloid leukemia-associated) (SET) similar protein (SETSIP) was indentified to be induced during somatic cell reprogramming. Importantly, when PiPS cells were treated with VEGF, SETSIP was translocated to the cell nucleus, directly bound to the VE-cadherin promoter, increasing vascular endothelial-cadherin (VE-cadherin) expression levels and EC differentiation. Functionally, PiPS-ECs improved neovascularization and blood flow recovery in a hindlimb ischemic model. Furthermore, PiPS-ECs displayed good attachment, stabilization, patency, and typical vascular structure when seeded on decellularized vessel scaffolds. These findings indicate that reprogramming of fibroblasts into ECs via SETSIP and VEGF has a potential clinical application.
Author	Karamariti, Eirini Margariti, Andriana Han, Jing-Dong J Zampetaki, Anna Baban, Dilair Ragoussis, Jiannis Zeng, Lingfang Tsai, Tsung-neng Huang, Yi Xu, Qingbo Hu, Yanhua Winkler, Bernhard
Author_xml	– sequence: 1 givenname: Andriana surname: Margariti fullname: Margariti, Andriana organization: Cardiovascular Division, King's College London British Heart Foundation Centre, London, United Kingdom – sequence: 2 givenname: Bernhard surname: Winkler fullname: Winkler, Bernhard – sequence: 3 givenname: Eirini surname: Karamariti fullname: Karamariti, Eirini – sequence: 4 givenname: Anna surname: Zampetaki fullname: Zampetaki, Anna – sequence: 5 givenname: Tsung-neng surname: Tsai fullname: Tsai, Tsung-neng – sequence: 6 givenname: Dilair surname: Baban fullname: Baban, Dilair – sequence: 7 givenname: Jiannis surname: Ragoussis fullname: Ragoussis, Jiannis – sequence: 8 givenname: Yi surname: Huang fullname: Huang, Yi – sequence: 9 givenname: Jing-Dong J surname: Han fullname: Han, Jing-Dong J – sequence: 10 givenname: Lingfang surname: Zeng fullname: Zeng, Lingfang – sequence: 11 givenname: Yanhua surname: Hu fullname: Hu, Yanhua – sequence: 12 givenname: Qingbo surname: Xu fullname: Xu, Qingbo
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SubjectTerms	Animals Antigens, CD - genetics Aorta - pathology Cadherins - genetics Cell Differentiation Cells, Cultured Cellular Reprogramming Endothelial Cells - cytology Fibroblasts - cytology Fibroblasts - metabolism Humans Induced Pluripotent Stem Cells - cytology Kruppel-Like Factor 4 Mice Mice, SCID Models, Genetic Neovascularization, Pathologic Promoter Regions, Genetic Stem Cells - cytology Stress, Mechanical Tissue Engineering - methods
Title	Direct reprogramming of fibroblasts into endothelial cells capable of angiogenesis and reendothelialization in tissue-engineered vessels
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