2007 Fiscal Year Final Research Report Summary
Molecular cloning and characterization of differentiation-inducing factors into the cardiac myocyte lineage using an expression cloning approach
| Project/Area Number |
18590773
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Kyoto University |
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Principal Investigator |
UEYAMA Tomomi Kyoto University, Dep. Experimental Therapeutics, Assistant Professor (80379388)
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| Co-Investigator(Kenkyū-buntansha) |
MATUBARA Hiroaki Kyoto Prefectural University of Medicine, Dep. Second Department of Medicine, Professor (10239072)
OH Hidemasa Kyoto University, Dep. Experiniental Therapeutics, Associate Professor (50372579)
HARADA Koichiro Kyoto University, Dep. Experiniental Therapeutics, Assistant Professor (30402902)
OGATA Takehiro Kyoto University, Dep. Experiniental Therapeutics, Research Associate (10402877)
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| Project Period (FY) |
2006 – 2007
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| Keywords | regenerative medicine / stem cells / cardiac myocyte / differentiation-inducing factor / cloning |
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| Research Abstract |
Since embryonic stem (ES) cells are capable of self-renew and differentiation into specialized cells in response to appropriate signals, ES cells are proposed as a promising source of functional cardiac myocytes for cardiac diseases. However, to date the in vitro differentiation of ES cells into cardiac myocytes remains inefficient and its molecular mechanisms are not fully elucidated. Therefore, the identification of differentiation-inducing factors into the cardiac myocyte lineage has become critical for understanding the molecular mechanisms of differentiation into cardiac myocyte and heart development, and facilitates therapeutic applications of ES cells in cardiac diseases. In the present study, we sought to identify differentiation-inducing factors into the cardiac myocyte lineage using an expression cloning approach. We induced differentiation of the mouse embryonic carcinoma cell line, P19CL cells, into cardac myocytes using DMSO. We made cDNA libraries using mRNA isolated from differentiating cells. We then generated recombinant retroviruses expressing the cDNA libraries. ES cell clones that express EGFP under the transcriptional control of a cardiac-specific α-MHC promoter were infected with recombinant retroviruses expressing the cDNA libraries. Gemonic DNA was extracted from differentiated cells into cardiac myocytes and performed PCR to isolate the integrated cDNAs into the genomic DNA. We obtained G protein beta polypeptide 2 like 1 (Gnb2I1, Rack1), oxidase assembly 1-like (Oxa1l), glutaredoxin 5 homolog (Glrx5), and aurora kinase A interacting protein 1(Aurkaip1) as candidate genes for differentiation-inducing factors into the cardiac myocyte lineage. We are studying their functions in differentiation of ES cells into cardiac myocyte. Our study will provide novel insights into the molecular mechanisms of cardiac myocyte differentiation and the development of novel approaches for the directed differentiation of ES cells into cardiac myocytes.
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[Journal Article] MURC, a muscle-restricted coiled-coil protein that modulates the Rho/ROCK pathway, induces cardiac dysfunction and conduction disturbance2008
Author(s)
Ogata, T., Ueyama, T., Isodono, K., Tagawa, M., Takehara, N., Kawashima, T., Harada, K., Takahashi, T., Shioi, T., Matsubara, H., Oh, H
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Journal Title
Mol Cell Biol 28
Pages: 3424-3436
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Skeletal muscle-derived progenitors capable of differentiating into cardiomyocytes proliferate through myostatin-independent TGF-beta family signaling2008
Author(s)
Nomura, T., Ueyama, T., Ashihara, E., Tateishi, K., Asada, S., Nakajima, N., Isodono, K., Takahashi, T., Matsubara, H., Oh, H
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Journal Title
Biochem Biophys Res Commun 365
Pages: 863-869
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Therapeutic potential of stem/progenitor cells in human skeletal muscle for cardiovascular regeneration2007
Author(s)
Nomura, T., Ashihara, E., Tateishi, K., Ueyama, T., Takahashi, T., Yamagishi, M., Kubo, T., Yaku, H., Matsubara, H., Oh, H
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Journal Title
Current Stem Cell Research and Therapy 2
Pages: 293-300
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Osteopontin is a myosphere-derived secretory molecule that promotes angiogenic progenitor cell proliferation through the phosphoinositide 3-kinase/Akt pathway2007
Author(s)
Ogata, T., Ueyama, T., Nomura, T., Asada, S., Tagawa, M., Nakamura, T., Takahashi, T., Matsubara, H., Oh, H
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Journal Title
Biochem Biophys Res Commun 359
Pages: 341-347
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Single cardiac stem cells require stem cell antigen-1 to proliferate and survive for efficient cardiovascular regeneration2007
Author(s)
Tateishi, K., Ashihara, E., Takehara, N., Nomura, T., Honsho, S., Nagagami, T., Morikawa, S., Takahashi, T., Ueyama, T., Matsubara, H., Oh, H
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Journal Title
J Cell Sci 120
Pages: 1791-800
Description
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[Journal Article] Overexpression of Toll-Like Receptors at the Vessel Wall Induces Atherosclerotic Lesion Formation. Synergism of TLR2 and TLR42007
Author(s)
Shinohara, M., Hirata, K., Yamashita, T., Takaya, T., Sasaki, N., Ueyama, T., Emoto, N., Inoue, N., Yokoyama, M., Kawashima, S
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Journal Title
Arterioscler Thromh Vasc Biol 11
Pages: 2384-2391
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Endogenous Smad2 Activation Modulates Cardiac Differentiation in Embryonic Stem Cells2007
Author(s)
Kitamura, R., Takahashi, T., Nakajima, N., Isodono, K., Asada, S., Ueno, H., Ueyama, T., Matsubara, H., Oh, H
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Journal Title
Circ Res 101
Pages: 78-87
Description
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