細胞分化機構から見た血管－神経共通シグナルの意義の解明

• Project Area: Vasculo-neural wiring and their interdependent crosstalk
• Project/Area Number: 23122509
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: 山下 潤 京都大学, iPS細胞研究所, 教授 (50335288)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000); Fiscal Year 2012: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2011: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: cAMP / 内皮細胞 / 細胞分化 / オピオイド / マイクロRNA / ES細胞 / サイクリックAMP / 分化

Outline of Annual Research Achievements

研究代表者はES細胞及びiPS細胞を用いた心血管分化再生研究を行ってきた。本研究は、「細胞分化の観点から血管と神経に共通な新しい分子群を同定し、血管、特に内皮細胞と神経分化の関連性に関して新たな理解をもたらす」ことを目的とし、１）オピオイドの内皮分化・血管形成における意義、２）cAMPシグナルと内皮・神経分化、３）Notch-beta-cateninシグナルと内皮・神経分化、４）新規動脈化誘導シグナルと神経分化、５）神経作用性物質の内皮分化における意義、の５項目の検討を行った。特に
１）に関しては、オピオイドの成体における意義を検討するため、カッパオピオイド受容体（KOR)欠損マウスにおける腫瘍血管新生を検討し、KOR欠損マウスにおいては腫瘍血管形成が亢進していることを見いだしている。
２）に関して、PKAがVEGF受容体発現を亢進させる機構の解析を行い、Ets転写因子群のEtv2遺伝子にcAMP応答配列が存在し、PKA下流で活性化されたCREBが同配列に結合し、Etv2発現を誘導することにより、種々のVEGF受容体遺伝子発現を誘導し、内皮分化を惹起することを明らかにした(Yamamizu, Stem Cells, 2012)。また、RSK4がPKAに結合してその活性を抑制することにより、血管内皮細胞の分化に対する新しいnegative regulatorとして働いていることを明らかにした。生体における血管発生過程においてもRSKは内皮分化抑制的に作用しており、RSKの作用を阻害剤により抑制すると血管内皮分化と血管形成の異常な促進が観察された。RSKの血管形成における新しい意義を明らかにした。また４）に関して、内皮細胞と神経細胞に共通して発現するmicroRNAを同定し、その標的分子の同定と機能確認を進めている。このように本研究において着実な進捗を認めている。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] PKA/CREB signaling triggers initiation of endothelial and hematopoietic cell differentiation via Etv2 induction (2012)
  • Author(s): Yamamizu K, Matsunaga T, Katayama S, Kataoka H, Takayama N, Eto K, Nishikawa SI, Yamashita JK
  • Journal Title: Stem Cells Volume: 30 Issue: 4 Pages: 687-696
  • DOI: 10.1002/stem.1041
  • Peer Reviewed
• [Journal Article] Protein Kinase A Determines Timing of Early Differentiation through Epigenetic Regulation with G9a (2012)
  • Author(s): Yamamizu K, Fujihara M, Tachibana M, Katayama S, Takahashi A, Hara E, Imai H, Shinkai Y, and Yamashita JK
  • Journal Title: Cell Stem Cell Volume: 10 Issue: 6 Pages: 759-770
  • DOI: 10.1016/j.stem.2012.02.022
  • Peer Reviewed
• [Journal Article] Pluripotent stem cell-engineered cell sheets re-assembled with defined cardiovascular populations ameliorate reduction in infarct heart function with cardiomyocyte-mediated neovascularization (2012)
  • Author(s): Masumoto H, Matsuo T, Yamamizu K, Uosaki H, Narazaki G, Katayama S, Marui A, Shimizu T, Ikeda T, Okano T, Sakata R, Yamashita JK
  • Journal Title: Stem Cells Volume: 30 Issue: 6 Pages: 1196-1205
  • DOI: 10.1002/stem.1089
  • Peer Reviewed
• [Journal Article] Protein kinase A determines timing of early differentiation through epigenetic regulation with G9a (2012)
  • Author(s): Yamamizu K, Fujihara M, Tachibana M, Katayama S, Takahashi A, Hara E, Imai H, Shinkai Y, Yamashita JK
  • Journal Title: Cell Stem Cell Volume: (in press)
  • Peer Reviewed
• [Journal Article] Effect of k-opioid receptor agonist on the growth of non-small cell lung cancer (NSCLC) cells (2012)
  • Author(s): Kuzumaki N, Suzuki A, Narita M, Hosoya T, Nagasawa A, Imai S, Yamamizu K, Morita H, Nagase H, Okada Y, Okano HJ, Yamashita JK, Okano H, Suzuki T, Narita M
  • Journal Title: Br J Cancer Volume: 106(Epub 2012 Feb 16) Issue: 6 Pages: 1148-1152
  • DOI: 10.1038/bjc.2011.574
  • Peer Reviewed
• [Journal Article] Potential of ES Cell Differentiation Culture for Vascular Biology (2012)
  • Author(s): Yamamizu K, Yamashita JK, Shin-Ichi Nishikawa
  • Journal Title: Handbook of Stem Cells Volume: (in press)
  • Peer Reviewed
• [Journal Article] Efficient and scalable purification of cardiomyocytes from human embryonic and induced pluripotent stem cells by VCAM1 surface (2011)
  • Author(s): Uosaki H
  • Journal Title: PLoS One Volume: 2011:6(8) Issue: 8 Pages: e23657-e23657
  • DOI: 10.1371/journal.pone.0023657
  • Peer Reviewed
• [Journal Article] The kappa opioid system regulates endothelial cell differentiation and pathfinding in vascular development (2011)
  • Author(s): Yamamizu K, Furuta S, Katayama S, Narita M, Kuzumaki N, Imai S, Nagase H, Suzuki T, Narita M, Yamashita JK
  • Journal Title: Blood(表紙) Volume: 118 Issue: 3 Pages: 775-785
  • DOI: 10.1182/blood-2010-09-306001
  • Peer Reviewed
• [Presentation] Diverse roles of protein kinase A signaling in early stage endothelial cell differentiation (Oral presentation). (2012)
  • Author(s): 山下 潤.
  • Organizer: 17th International Vascular Biology Meeting. ‘The highest scored abstract’
  • Place of Presentation: Wiesbaden, Germany.
  • Year and Date: 2012-06-04
  • Invited
• [Presentation] .Differentiation Stage-specific Roles of Protein Kinase A Signaling in Endothelial Cell Lineages. (2012)
  • Author(s): Yamashita JK
  • Organizer: 2012 KAIST Mini-symposium. Stem cells and Vascular Biology.
  • Place of Presentation: Daejeon. Korea
  • Invited
• [Presentation] Fate determination processes of endothelial lineages regulated by protein kinase A signaling in Pluripotent stem cell differentiation. (2012)
  • Author(s): Yamashita JK.
  • Organizer: Japan-Korea Joint Symposium of Vascular Biology.
  • Place of Presentation: Tokushima, Japan.
  • Invited
• [Presentation] ES/iPS細胞とケミカルバイオロジー-新しい心臓再生治療における低分子化合物のポテンシャル- (2011)
  • Author(s): 山下潤
  • Organizer: 早稲田大学先進理工学部講演会
  • Place of Presentation: 東京(招待講演)
  • Year and Date: 2011-12-20
• [Presentation] RSK4 acts as a novel and negative regulator in endothelial ceU differentiation via cAMP/PKA dependent pathway (2011)
  • Author(s): Matsunaga T, Koshiba-Takeuchi K, Kojima M, Takeuchi JK, Yamashita JK
  • Organizer: The 19th Annual Meeting of the Japanese vascular and medicine organization
  • Place of Presentation: Tokyo, Japan
  • Year and Date: 2011-12-10
• [Presentation] RSK4によるcAMP/PKAシグナルを介した血管内皮細胞の分化制御 (2011)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第15回日本心血管内分泌代謝学会
  • Place of Presentation: 大阪
  • Year and Date: 2011-11-25
• [Presentation] ES・iPS細胞を用いた心臓再生の試み (2011)
  • Author(s): 山下潤
  • Organizer: 第15回日本心不全学会シンポジウム「心不全における再生治療up to date」
  • Place of Presentation: 鹿児島(招待講演)
  • Year and Date: 2011-10-14
• [Presentation] 多能性幹細胞由来心臓構成細胞の再構成による心臓組織シート-治療効果と機能回復メカニズム解析- (2011)
  • Author(s): 升本英利, 池田義, 坂田隆造
  • Organizer: 第64回日本胸部外科学会定期学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 2011-10-11
• [Presentation] G9aによるヒストンメチル化を介したPKAの細胞分化速度制御機構の解明 (2011)
  • Author(s): 山水康平, 藤原摩耶子, 立花誠, 片山志織, 今井裕, 眞貝洋一, 山下潤
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2011-09-22
• [Presentation] Central Role of Cardiomyocytes in Cell Therapy : Transplantation of Cardiac Tissue Sheets with Defined Cell Populations from Pluripotent Stem Cells (2011)
  • Author(s): Masumoto H, Shimizu T, Ikeda T, Okano T, Sakata R, Yamashita JK
  • Organizer: European Society of Cardiology (ESC) Congress
  • Place of Presentation: Paris, France
  • Year and Date: 2011-08-30
• [Presentation] Cardiovascular regeneration with ES & iPS cells (2011)
  • Author(s): Yamashita JK
  • Organizer: The 8th Korea-Japan Joint Symposium of Vascular Biology
  • Place of Presentation: Busan, Korea(invited)
  • Year and Date: 2011-08-25
• [Presentation] Embryonic Stem (ES) Cell-engineered Tissue Sheets with Defined Cardiac Cell Populations Ameliorate Function after Myocardial Infarction (2011)
  • Author(s): 升本英利, 清水達也, 池田義, 岡野光夫, 坂田隆造, 山下潤
  • Organizer: 第75回日本循環器学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2011-08-03
• [Presentation] VCAM1 as a Specific Marker for Cardiomyocytes from Human Embryonic and induced Pluripotent Stem Cells (2011)
  • Author(s): Uosaki H, Yamashita JK
  • Organizer: Bassetlaw Community and Voluntary Service
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2011-07-19
• [Presentation] Cardiomyocyte Proliferating Chemicals : Activation of Proliferation of ESC/iPSC-derived Cardiomyocytes (2011)
  • Author(s): Uosaki H, Yamashita JK
  • Organizer: Bassetlaw Community and Voluntary Service
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2011-07-18
• [Presentation] 多能性幹細胞由来心臓組織シートによる虚血性心疾患治療への挑戦 (2011)
  • Author(s): 升本英利, 池田義, 坂田隆造, 山下潤
  • Organizer: 第9回メタボリックシンドロームカンファレンス
  • Place of Presentation: 京都
  • Year and Date: 2011-07-16
• [Presentation] 心臓再生へ向けた多面的アプローチ～ES/iPS細胞を用いた創薬研究と細胞治療～ (2011)
  • Author(s): 山下潤
  • Organizer: 文部科学省私立大学学術高度化推進事業ハイテクリサーチセンター整備事業・第4回シンポジウム
  • Place of Presentation: 東大阪(招待講演)
  • Year and Date: 2011-06-25
• [Presentation] Mouse embryonic stem cell-engineered tissue sheets with defined cardiac cell populations ameliorate function after myocardial infarction (2011)
  • Author(s): Masumoto H, Ikeda T, Sakata R, Yamashita JK
  • Organizer: The 9th International Society for Stem Cell Research (ISSCR)
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2011-06-17
• [Presentation] マウス多能性幹細胞由来心臓組織シートの心筋梗塞後リモデリング抑制効果 (2011)
  • Author(s): 升本英利, 池田義, 清水達也, 岡野光夫, 坂田隆造, 山下潤
  • Organizer: 第32回日本炎症・再生医学会
  • Place of Presentation: 京都
  • Year and Date: 2011-06-11
• [Presentation] ES/iPS細胞を用いた再生医-心血管再生を中心に- (2011)
  • Author(s): 山下潤
  • Organizer: 神戸薬科大学第37回卒後研修講座
  • Place of Presentation: 神戸(招待講演)
  • Year and Date: 2011-06-05
• [Presentation] iPS細胞の基礎と応用 (2011)
  • Author(s): 山下潤
  • Organizer: 再生医療学会エデュケーショナルセミナー
  • Place of Presentation: 東京(招待講演)
  • Year and Date: 2011-05-31
• [Presentation] 心臓再生へ向けた多面的アプローチ～ES/iPSiB胞を用いた細胞治療と創薬研究～ (2011)
  • Author(s): 山下潤
  • Organizer: 第24回北摂循環器研究会
  • Place of Presentation: 吹田(招待講演)
  • Year and Date: 2011-05-24
• [Presentation] Development of cardiac regeneration strategies with pluripotent stem cells (2011)
  • Author(s): Yamashita JK
  • Organizer: JST/CIRM Workshop. "Early Translational Research on Stem Cells"
  • Place of Presentation: Kobe, Japan(invited)
  • Year and Date: 2011-05-16
• [Presentation] Endothelial and hematopoietic cell differentiation are initiated by PKA/CREB pathway via Etv2 induction (2011)
  • Author(s): Yamamizu K, Matsunaga T, Katayama S, Yamashita JK
  • Organizer: The 19th Annual Meeting of the Japanese vascular and medicine organization
  • Place of Presentation: Tokyo, Japan
• [Presentation] Identification of a novel microRNA regulating angiogenic responses (2011)
  • Author(s): Doi K, Matsunaga T, Yamamizu K, Fukushima H, Miyamoto S, Yamashita JK
  • Organizer: The 19th Annual Meeting of the Japanese vascular and medicine organization
  • Place of Presentation: Tokyo, Japan
• [Presentation] PKA-CREB pathway initiates endothelial and hematopoietic cell differentiation through Etv2 induction (2011)
  • Author(s): Yamamizu K, Matsunaga T, Katayama S, Yamashita JK
  • Organizer: American Heart Association Scientific Session
  • Place of Presentation: Orland, USA
• [Presentation] PKAによるヒストンメチル化酵素G9aを介した細胞分化速度制御機構の解明 (2011)
  • Author(s): 山水康平, 藤原摩耶子, 立花誠, 片山志織, 今井裕, 眞貝洋一, 山下潤
  • Organizer: 第2回Molecular Cardiovascular Conference II
  • Place of Presentation: 小樽
• [Presentation] PKA-CREB pathway initiates endothelial and hematopoietic cell differentiation through Etv2 induction (2011)
  • Author(s): Yamamizu K, Matsunaga T, Katayama S, Yamashita JK
  • Organizer: The 9th Japan-Korea Joint Symposium on Vascular Biology
  • Place of Presentation: Busan, Korea
• [Presentation] Protein kinase A accelerates early programming from mouse ES cells through epigenetic inhibition of Oct3/4 and Nanog expression (2011)
  • Author(s): Yamamizu K, Katayama S, Tachibana M, Shinkai Y, Yamashita JK
  • Organizer: The 9th International Society for Stem cell Research
  • Place of Presentation: Toronto, Canada
• [Presentation] Identification of novel microRNA regulating angiogenic response (2011)
  • Author(s): Doi K, Matsunaga T, Yamamizu K, Miyamoto S, Yamashita JK
  • Organizer: The 9th International Society for Stem cell Research
  • Place of Presentation: Toronto, Canada
• [Presentation] 血管形成機構および血管走行制御におけるオピオイドペプチドの役割 (2011)
  • Author(s): 山水康平, 古田貞由, 片山志織, 成田道子, 葛巻直子, 今井哲司, 長瀬博, 鈴木勉, 成田年, 山下潤
  • Organizer: 第84回日本内分泌学会学術総会
  • Place of Presentation: 神戸
• [Remarks] 山下研究室ホームページ
  • URL: http://www.frontier.kyoto-u.ac.jp/es02/index.htm
• [Patent(Industrial Property Rights)] The method of producing the group of cells containing cardiomyocytes and vascular tissue cells (2012)
  • Inventor(s): 山下潤/升本英利
  • Industrial Property Rights Holder: 京都大学
  • Filing Date: 2012-03-15
  • Overseas