Development of a novel vascular-targeting therapy through stem cell research

• Project/Area Number: 24390197
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: Kyoto University
• Principal Investigator: YAMASHITA Jun 京都大学, iPS細胞研究所, 教授 (50335288)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2014: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2013: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2012: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
• Keywords: 血管新生 / 多能性幹細胞 / 内皮細胞 / 分化 / 血管内皮増殖因子 / アンジオポイエチン / 幹細胞 / 血管形成 / マイクロRNA / シグナル / キナーゼ

Outline of Final Research Achievements

Purpose of this study is that development of novel therapies for ischemic heart disease or cancer through manipulating vascular formation using advanced pluripotent stem cell research.
We demonstrated that kappa-opioid receptor suppressed tumor angiogenesis through inhibiting expression of type II vascular endothelial growth factor receptor. In addition, we reported a novel function of angiopoietin-1 in coronary vein formation as a collaboration with Department of cardiovascular medicine, University of Tokyo.

Research Products

• [Journal Article] Single-step generation of gene knockout-rescue system in pluripotent stem cells by promoter insertion with CRISPR/Cas9 (2014)
  • Author(s): Matsunaga T, Yamashita JK.*
  • Journal Title: Biochem Biophys Res Commun Volume: 444 Pages: 158-163
  • Peer Reviewed / Open Access
• [Journal Article] Dual modulation of the mitochondrial permeability transition pore and redox signaling synergistically promotes cardiomyocyte differentiation from pluripotent stem cells. (2014)
  • Author(s): Cho SW, Park JS, Heo HJ, Park SW, Song S, Kim I, Han YM, Yamashita JK, Youm JB, Han J, Koh GY.
  • Journal Title: J Am Heart Assoc Volume: 3(2) Issue: 2
  • DOI: 10.1161/jaha.113.000693
  • Peer Reviewed / Open Access
• [Journal Article] Myocardium-derived angiopoietin-1 is essential for coronary vein formation in the developing heart (2014)
  • Author(s): Arita Y, Nakaoka Y, Matsunaga T, Kidoya H, Yamamizu K, Arima Y, Kataoka-Hashimoto T, Ikeoka K, Yasui T, Masaki T, Yamamoto K, Higuchi K, Park JS, Shirai M, Nishiyama K, Yamagishi H, Otsu K, Kurihara H, Minami T, Yamauchi-Takihara K, Koh GY, Mochizuki N, Takakura N, Sakata Y, Yamashita JK, Komuro I.
  • Journal Title: Nat Commun Volume: 5 Pages: 4552-4552
  • Peer Reviewed / Open Access
• [Journal Article] Human iPS cell-engineered cardiac tissue sheets with cardiomyocytes and vascular cells for cardiac regeneration. (2014)
  • Author(s): Masumoto H, Ikuno T, Takeda M, Fukushima H, Marui A, Katayama S, Shimizu T, Ikeda T, Okano T, Sakata R, Yamashita JK.
  • Journal Title: Sci Rep Volume: 4 Issue: 1 Pages: 6716-6716
  • DOI: 10.1038/srep06716
  • Peer Reviewed / Open Access
• [Journal Article] Single-step generation of gene knockout-rescue system in pluripotent stem cells by promoter insertion with CRISPR/Cas9. (2014)
  • Author(s): Matsunaga T, Yamashita JK*
  • Journal Title: Biochem Biophys Res Commun Volume: 444 Issue: 2 Pages: 158-163
  • DOI: 10.1016/j.bbrc.2014.01.037
  • Peer Reviewed
• [Journal Article] Kappa-Opioids inhibit tumor angiogenesis by suppressing VEGF signaling. (2013)
  • Author(s): Yamamizu K, Furuta S, Hamada Y, Yamashita A, Kuzumaki N, Narita M, Doi K, Katayama S, Nagase H, Yamashita JK, Narita M.
  • Journal Title: Scientific Reports Volume: 3 Issue: 1 Pages: 3213-3213
  • DOI: 10.1038/srep03213
  • Peer Reviewed / Open Access
• [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] Multiple analyses of G-protein coupled receptor (GPCR) expression in the development of gefitinib-resistance in transforming non-small-cell lung cancer. (2012)
  • Author(s): Kuzumaki N
  • Journal Title: PLoS One Volume: 7 Issue: 10 Pages: e44368-e44368
  • DOI: 10.1371/journal.pone.0044368
  • 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, 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] 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
• [Presentation] 2型VEGF受容体陽性細胞純化再培養はヒトiPS細胞からの血管内皮細胞分化誘導を効率化する. (2015)
  • Author(s): 幾野 毅
  • Organizer: 第18回日本再生医療学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2015-03-19
• [Presentation] 2型VEGF受容体陽性細胞純化再培養によるヒトiPS細胞からの効率的血管内皮細胞分化誘導. (2015)
  • Author(s): 幾野 毅
  • Organizer: 第45回日本心臓血管外科学会学術総会.
  • Place of Presentation: 京都
  • Year and Date: 2015-02-18
• [Presentation] Scalable and highly efficient endothelial cell differentiation methods from human induced pluripotent stem cells based on monolayer and serum-free culture for realization of regenerative medicine. (2015)
  • Author(s): 幾野 毅
  • Organizer: The 18th Takeda Science Foundation Symposium on Bioscience,
  • Place of Presentation: 大阪
  • Year and Date: 2015-01-15 – 2015-01-17
• [Presentation] ２型血管内皮細胞増殖因子受容体陽性細胞の純化再培養によるヒトiPS 細胞からの血管内皮細胞分化誘導の効率化 (2014)
  • Author(s): 幾野 毅
  • Organizer: 第67回日本胸部外科学会総会
  • Place of Presentation: 福岡
  • Year and Date: 2014-10-03
• [Presentation] Scalable and highly efficient endothelial cell differentiation methods from human induced pluripotent stem cells for realization of regenerative medicine． (2014)
  • Author(s): 幾野 毅
  • Organizer: 第5回Molecular Cardiovascular ConferenceⅡ.
  • Place of Presentation: Kobe
  • Year and Date: 2014-09-06
• [Presentation] ヒトiPS細胞由来心筋特異的前駆細胞の同定 (2014)
  • Author(s): 武田 匡史、山下 潤
  • Organizer: 第5回 Molecular Cardiovascular Conference Ⅱ
  • Place of Presentation: 神戸
  • Year and Date: 2014-09-05 – 2014-09-06
• [Presentation] Scalable and highly efficient endothelial cell differentiation methods from human induced pluripotent stem cells based on monolayer and serum-free culture. (2014)
  • Author(s): Ikuno T.
  • Organizer: ESC Congress2014.
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2014-08-31
• [Presentation] Scalable and highly efficient endothelial cell differentiation methods from human induced pluripotent stem cells based on monolayer and serum-free culture for realization of regenerative medicine. (2014)
  • Author(s): Ikuno T.
  • Organizer: International Society for Stem Cell Research (ISSCR) 12th Annual Meeting.
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2014-06-20
• [Presentation] Identification of novel chemicals potently inducing cardiomyocyte differentiation from mouse and human pluripotent stem cells (2014)
  • Author(s): Fukushima H.
  • Organizer: ISSCR (International Society for Stem Cell Research) 12th Annual Meeting
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2014-06-18 – 2014-06-21
• [Presentation] Identification and application of human iPS derived-cardiac progenitor enriched population for cardiac regeneration. (2014)
  • Author(s): Takeda T, Yamashita JK.
  • Organizer: The 3ird Retreat of Center for iPS Cell Research and Application (CiRA)
  • Place of Presentation: Shiga
  • Year and Date: 2014-04-19
• [Presentation] Yamashita JK. Fate determination processes of endothelial lineages regulated by protein kinase A signaling in Pluripotent stem cell differentiation. (2012)
  • Author(s): K. Yamashita
  • Organizer: Japan-Korea Joint Symposium of Vascular Biology.
  • Place of Presentation: Tokushima, Japan.(招待講演)
  • Year and Date: 2012-12-06
• [Presentation] RSK4 is a novel regulator of endothelial cells differentiation through direct inhibition of Protein kinase A activity. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第20回日本血管生物医学会学術大会.
  • Place of Presentation: 徳島
  • Year and Date: 2012-12-06
• [Presentation] Differentiation Stage-specific Roles of Protein Kinase A Signaling in Endothelial Cell Lineages. (2012)
  • Author(s): K. Yamashita
  • Organizer: 2012 KAIST Mini-symposium. Stem cells and Vascular Biology.
  • Place of Presentation: Daejeon. Korea.(招待講演)
  • Year and Date: 2012-10-22
• [Presentation] 血管内皮分化におけるRSK4の役割. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第3回Molecular Cardiovascular Conference II.
  • Place of Presentation: 小樽
  • Year and Date: 2012-09-08
• [Presentation] RSK4 is a Novel Negative Regulator of Endothelial Differentiation via Direct Suppression of Protein Kinase A Activity (poster). BCVS (Basic Cardiovascular Sciences) (2012)
  • Author(s): Matsunaga T, Jun K. Yamashita
  • Organizer: 2012 Scientific Sessions, AHA (American Heart Association).
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2012-07-23
• [Presentation] RSK4 is a novel negative regulator of endothelial differentiation via direct suppression of cAMP/PKA signaling. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第10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: 2012-07-14
• [Presentation] RSK4 regulates endothelial differentiation via inhibition of cAMP/PKA-mediated Flk1 induction. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第33回日本炎症・再生医学会
  • Place of Presentation: 福岡
  • Year and Date: 2012-07-05
• [Presentation] RSK4 regulates endothelial differentiation through suppression of cAMP/PKA-mediated Flk1 induction (poster) (2012)
  • Author(s): Matsunaga T, Jun K. Yamashita
  • Organizer: 17th International Vascular Biology Meeting.
  • Place of Presentation: Wiesbaden, Germany.
  • Year and Date: 2012-06-04
• [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.
  • Place of Presentation: Wiesbaden, Germany.(招待講演)
  • Year and Date: 2012-06-04
• [Presentation] RSK4によるcAMP/PKAシグナルを介した新しい内皮細胞分化制御機構. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第85回日本内分泌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2012-04-20
• [Presentation] cAMP/PKA signalingを介したRSK4による内皮細胞分化制御機構. (2012)
  • Author(s): 松永太一, 山下潤
  • Organizer: 第49回日本臨床分子医学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 2012-04-13
• [Presentation] RSK4 suppresses endothelial cell fate through direct inhibition of PKA activity. (poster)
  • Author(s): Matsunaga T.
  • Organizer: ISSCR (International Society for Stem Cell Research) 11th Annual Meeting
  • Place of Presentation: Boston, USA.
• [Presentation] 内皮細胞の分化制御機構におけるRSK4の役割
  • Author(s): 松永 太一
  • Organizer: 第33回日本炎症・再生医学会
  • Place of Presentation: 京都
• [Presentation] RSK4による内皮細胞分化の新規抑制機構
  • Author(s): 松永 太一
  • Organizer: 第4回Molecular Cardiovascular Conference II
  • Place of Presentation: 小樽
• [Presentation] 内皮細胞分化におけるRSK4の役割
  • Author(s): 松永 太一
  • Organizer: 第21回日本血管生物医学会学術大会
  • Place of Presentation: 豊中
• [Presentation] Crispr/Cas9を用いたプロモーター配列挿入による簡便な誘導性遺伝子過剰発現系の構築とその幹細胞研究応用
  • Author(s): 松永 太一
  • Organizer: 第13回日本再生医療学会
  • Place of Presentation: 京都
• [Presentation] Efficient and Scalable Endothelial Cell Differentiation Method from Human Induced Pluripotent Stem Cells Based on 2-D Monolayer and Serum-Free Culture.
  • Author(s): Ikuno T, Masumoto H, Marui A, Ikeda T, Sakata R, Yamashita JK.
  • Organizer: ISSCR(International Society for Stem Cell Research) 11th Annual Meeting
  • Place of Presentation: Boston, USA.
• [Presentation] ヒト人工多能性幹(iPS)細胞からの効率的な内皮細胞分化誘導法
  • Author(s): 幾野 毅
  • Organizer: 第34回日本炎症再生医学会
  • Place of Presentation: 京都
• [Presentation] Efficient endothelial cell differentiation protocol from human induced pluripotent stem cells based on monolayer and serum-free culture for realization of vascular regenerative medicine．Rapid Fire - Remodelling the vessel wall.
  • Author(s): Ikuno T, Masumoto H, Marui A, Ikeda T, Sakata R, Yamashita JK.
  • Organizer: ESC Congress2013
  • Place of Presentation: Amsterdam, Netherlands.
• [Presentation] ヒト人工多能性幹（iPS）細胞からの効率的かつ高収量な内皮細胞分化誘導法
  • Author(s): 幾野 毅
  • Organizer: 第4回Molecular Cardiovascular ConferenceII
  • Place of Presentation: 北海道
• [Presentation] 無血清二次元単層培養によるヒトiPS細胞からの高効率高収量な内皮細胞分化誘導法
  • Author(s): 幾野 毅
  • Organizer: 第21回日本血管生物医学会学術集会
  • Place of Presentation: 大阪
• [Presentation] ヒトiPS細胞を用いた二次元単層培養による効率的内皮細胞分化誘導法
  • Author(s): 幾野 毅
  • Organizer: 第66回日本胸部外科学会定期学術集会
  • Place of Presentation: 仙台
• [Presentation] ES細胞/iPS細胞を用いた心血管分化研究とその応用
  • Author(s): 山下 潤
  • Organizer: 安全性評価研究会（特別講演）
  • Place of Presentation: 大阪
• [Presentation] ES細胞・iPS細胞を用いた心血管分化再生研究
  • Author(s): 山下 潤
  • Organizer: 愛知Rising Force Meeting（特別講演）
  • Place of Presentation: 名古屋
  • Invited
• [Presentation] ES細胞/iPS細胞を用いた多面的心臓再生治療戦略の開発
  • Author(s): 山下 潤
  • Organizer: 岐阜心不全研究会（特別講演）
  • Place of Presentation: 岐阜
  • Invited
• [Presentation] Multiple and integrative approaches to cardiovascular diseases with stem cell technology.
  • Author(s): Yamashita JK.
  • Organizer: International Conference on Endothelin 2013 (Special Guest Session)
  • Place of Presentation: 東京
  • Invited
• [Presentation] Vascular cell differentiation and tissue regeneration.
  • Author(s): Yamashita JK.
  • Organizer: 第８６回日本生化学会国際シンポジウム「血管・リンパ管の発生・形成機構とシグナル伝達」
  • Place of Presentation: 横浜
  • Invited
• [Presentation] 多能性幹細胞からの内皮細胞分化とその応用
  • Author(s): 山下 潤
  • Organizer: 日本血管生物医学会シンポジウム
  • Place of Presentation: 大阪
  • Invited
• [Presentation] ES細胞・iPS細胞を用いた多面的心臓再生治療戦略の開発
  • Author(s): 山下 潤
  • Organizer: 奈良県立医科大学祭シンポジウム
  • Place of Presentation: 橿原
  • Invited
• [Presentation] 多能性幹細胞を用いた多面的心血管再生治療戦略の開発
  • Author(s): 山下 潤
  • Organizer: 日本心血管内分泌代謝学会シンポジウム
  • Place of Presentation: 大阪
  • Invited
• [Presentation] 多能性幹細胞を用いた心血管分化再生に関する多面的研究
  • Author(s): 山下 潤
  • Organizer: 徳島脈管研究会（特別講演）
  • Place of Presentation: 徳島
  • Invited
• [Remarks] 山下研究室のホームページ
  • URL: http://www.cira.kyoto-u.ac.jp/yamashita/achievement.html
• [Remarks] 山下研究室｜京都大学iPS細胞研究所／再生医科学研究所
  • URL: http://www.frontier.kyoto-u.ac.jp/es02/index.htm
• [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)] 効率的な内皮細胞の誘導方法 (2013)
  • Inventor(s): 山下潤、幾野毅、松永太一
  • Industrial Property Rights Holder: 国立大学法人京都大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-116428
  • Filing Date: 2013-05-31