Epithelial-mesenchymal interaction in the tras-differentiation from mouse epidermal cells to corneal epithelial cells

• Project/Area Number: 17K11484
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: Ehime University
• Principal Investigator: Shiraishi Atsushi 愛媛大学, 医学系研究科, 教授 (90314963)
• Co-Investigator(Kenkyū-buntansha): 小林 剛 愛媛大学, 医学系研究科, 寄附講座講師 (70380285) ; 三谷 亜里沙 愛媛大学, 医学部附属病院, 医員 (60648096)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 角膜上皮 / 表皮細胞 / 形質転換 / 角膜輪部 / 角膜 / 再生医療 / 上皮細胞培養

Outline of Final Research Achievements

In the present study, we employed three-dimensional culture model including mouse epidermal sp cells, human corneal stromal fibloblasts, and human corneal limbal stromal fibloblasts and demonstrated the quantitive analysis of corneal epithelial differentiation markers expressed in the mouse epidermal sp cells. Furthermore, to elucidate the effect of corneal limbal stromal factors including Wnt related factors for trans-differentiation from mouse epidermal cells to corneal epithelial cells, we performed tras-criptome analysis of corneal limbal stromal fibloblasts. We analyzed candidate factors that can be related to trans-differentiation to the corneal epithelial cells, and found 21 secreted proteins and 15 extracellular matrices genes.

Academic Significance and Societal Importance of the Research Achievements

本研究で得られた成果は、形質転換および角膜上皮の分化誘導における分子機構の解明を促進し、表皮幹細胞を用いた角膜上皮再生医療の実現に貢献できると考えられる。さらに、人工多能性幹細胞（iPS細胞）等を用いた次世代の再生医療の実現に向けても、本研究で得られる知見が利用可能であり、当分野において多大な貢献ができると考えられる。

Research Products

• [Journal Article] Characterization of doxycycline-dependent inducible Simian Virus 40 large T antigen immortalized human conjunctival nepithelial cell line. (2019)
  • Author(s): Mitani A, Kobayashi T, Hayashi Y, Matsushita N, Matsushita S, Nakao S, Takahira N, Shiraishi A, Ohashi Y.
  • Journal Title: PLoS One Volume: 14 Issue: 9 Pages: e0222454-e0222454
  • DOI: 10.1371/journal.pone.0222454
  • Peer Reviewed / Open Access
• [Journal Article] Retrospective Observational Study on Rebamipide Ophthalmic Suspension on Quality of Life of Dry Eye Disease Patients. (2019)
  • Author(s): Sakane Y, Yamaguchi M, Shiraishi A.
  • Journal Title: J Ophthalmol. Volume: May 2 Pages: 8145731-8145731
  • DOI: 10.1155/2019/8145731
  • Peer Reviewed / Open Access
• [Journal Article] Histone H3K27 Demethylase Negatively Controls the Memory Formation of Antigen-Stimulated CD8+ T Cells (2019)
  • Author(s): Yamada Takeshi、Nabe Shogo、Toriyama Koji、Suzuki Junpei、Inoue Kazuki、Imai Yuuki、Shiraishi Atsushi、Takenaka Katsuto、Yasukawa Masaki、Yamashita Masakatsu
  • Journal Title: The Journal of Immunology Volume: 202 Issue: 4 Pages: 1088-1098
  • DOI: 10.4049/jimmunol.1801083
  • Peer Reviewed / Open Access
• [Journal Article] In Vivo Confocal Microscopic Observations of Vortex Keratopathy in Patients with Amiodarone-Induced Keratopathy and Fabry Disease. (2018)
  • Author(s): Ikegawa Y, Shiraishi A, Hayashi Y, Ogimoto A, Ohashi Y
  • Journal Title: J Ophthalmol. Volume: 20 Pages: 5315137-5315137
  • DOI: 10.1155/2018/5315137
  • Peer Reviewed / Open Access
• [Journal Article] Effect of Botulinum Toxin A Treatment on Eyelid Pressure in Eyes with Blepharospasm. (2018)
  • Author(s): Namiguchi K, Mizoue S, Ohta K, Shiraishi A.
  • Journal Title: Curr Eye Res. Volume: Epub ahead of print Issue: 7 Pages: 896-901
  • DOI: 10.1080/02713683.2018.1464191
  • Peer Reviewed
• [Journal Article] Generation and Characterization of a Novel Mouse Line, Keratocan-rtTA (KeraRT), for Corneal Stroma and Tendon Research. (2017)
  • Author(s): Zhang Y, Kao WW, Hayashi Y, Zhang L, Call M, Dong F, Yuan Y, Zhang J, Wang YC, Yuka O, Shiraishi A, Liu CY.
  • Journal Title: Invest Ophthalmol Vis Sci. Volume: 58 Issue: 11 Pages: 4800-4808
  • DOI: 10.1167/iovs.17-22661
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mechanism of Proliferation of Cultured Human Corneal Endothelial Cells. (2017)
  • Author(s): Joko T, Shiraishi A, Kobayashi T, Ohashi Y, Higashiyama S.
  • Journal Title: Cornea. Volume: Suppl Issue: 1 Pages: S41-S45
  • DOI: 10.1097/ico.0000000000001337
  • Peer Reviewed
• [Journal Article] Effect of Mandarin Orange Yogurt on Allergic Conjunctivitis Induced by Conjunctival Allergen Challenge. (2017)
  • Author(s): Hara Y, Shiraishi A, Sakane Y, Takezawa Y, Kamao T, Ohashi Y, Yasunaga S, Sugahara T.
  • Journal Title: Invest Ophthalmol Vis Sci. Volume: 58 Issue: 7 Pages: 2922-2929
  • DOI: 10.1167/iovs.16-21206
  • Peer Reviewed
• [Journal Article] Observation of Retrocorneal Plaques in Patients With Infectious Keratitis Using Anterior Segment Optical Coherence Tomography. (2017)
  • Author(s): Takezawa Y, Suzuki T, Shiraishi A.
  • Journal Title: Cornea. Volume: 36 Issue: 10 Pages: 1237-1242
  • DOI: 10.1097/ico.0000000000001286
  • Peer Reviewed