Functional analysis in the transcriptional network in human corneal epithelial cell

• Project/Area Number: 16K20324
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Ophthalmology
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: Kitazawa Koji 京都府立医科大学, 医学(系)研究科(研究院), 特任助教 (10760803)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: OVOL2 / PAX6 / 角膜上皮 / 転写因子 / リプログラミング / 分化 / 細胞運命 / KLF4 / コア転写因子 / ダイレクトリプログラミング

Outline of Final Research Achievements

Functional analysis of core transcription factor in human corneal epithelial cells (CECs) revealed that CECs with OVOL2 knockdown became fibroblast-like cells, and OVOL2 knockdown reduced the expression of CEC-specific genes including keratin (K) 12 and E-cadherin, and the barrier function of CECs as measured by transepithelial electrical resistance. Next, when PAX6, which is an important transcription factor for eye development, was depleted by CRISPR/Cas9 targeted for PAX6 in human CECs, CEC-related gene including K12 and K3, was down-regulated, while K1 and K10, which were skin-related genes were up-regulated. These results suggest that PAX6 and OVOL2 were required for the terminal differentiation of CECs and, maintained the CEC identity.

Research Products

• [Journal Article] Topical ganciclovir treatment post-Descemet’s stripping automated endothelial keratoplasty for patients with bullous keratopathy induced by cytomegalovirus (2018)
  • Author(s): Kitazawa Koji、Jongkhajornpong Passara、Inatomi Tsutomu、Koizumi Noriko、Kayukawa Kanae、Wakimasu Koichi、Sotozono Chie、Kinoshita Shigeru
  • Journal Title: British Journal of Ophthalmology Volume: 印刷中 Issue: 9 Pages: 1293-1297
  • DOI: 10.1136/bjophthalmol-2017-311145
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Long-Term Outcome Post Penetrating Keratoplasty in a Pedigree with the G177E Mutation in the UBIAD1 Gene for Schnyder Corneal Dystrophy. (2018)
  • Author(s): Kitazawa K, Wakimasu K, Kayukawa K, Sugimoto M, Nakai J, Weiss J, Ueno M, Sotozono C, Kinoshita S
  • Journal Title: Cornea Volume: 37 Issue: 5 Pages: 554-559
  • DOI: 10.1097/ico.0000000000001511
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The existence of dead cells in donor corneal endothelium preserved with storage media (2017)
  • Author(s): Kitazawa Koji、Inatomi Tsutomu、Tanioka Hidetoshi、Kawasaki Satoshi、Nakagawa Hiroko、Hieda Osamu、Fukuoka Hideki、Okumura Naoki、Koizumi Noriko、Iliakis Bernie、Sotozono Chie、Kinoshita Shigeru
  • Journal Title: British Journal of Ophthalmology Volume: 101 Issue: 12 Pages: 1725-1730
  • DOI: 10.1136/bjophthalmol-2017-310913
  • Peer Reviewed
• [Journal Article] Predictive clinical factors of cystoid macular edema in patients with Descemet’s stripping automated endothelial keratoplasty (2017)
  • Author(s): Kitazawa Koji、Kayukawa Kanae、Wakimasu Koichi、Yokota Isao、Inatomi Tsutomu、Hieda Osamu、Mori Kazuhiko、Sotozono Chie、Kinoshita Shigeru
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 7412-7412
  • DOI: 10.1038/s41598-017-07079-x
  • Peer Reviewed / Open Access
• [Journal Article] Elastic microfibril distribution in the cornea: Differences between normal and keratoconic stroma (2017)
  • Author(s): White Tomas L.、Lewis Philip N.、Young Robert D.、Kitazawa Koji、Inatomi Tsutomu、Kinoshita Shigeru、Meek Keith M.
  • Journal Title: Experimental Eye Research Volume: 159 Pages: 40-48
  • DOI: 10.1016/j.exer.2017.03.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Safety of anterior chamber paracentesis using a 30-gauge needle integrated with a specially designed disposable pipette (2017)
  • Author(s): Kitazawa Koji、Sotozono Chie、Koizumi Noriko、Nagata Kenji、Inatomi Tsutomu、Sasaki Hiroshi、Kinoshita Shigeru
  • Journal Title: British Journal of Ophthalmology Volume: 101 Issue: 5 Pages: 548-550
  • DOI: 10.1136/bjophthalmol-2016-309650
  • Peer Reviewed
• [Journal Article] A case of fungal keratitis and endophthalmitis post penetrating keratoplasty resulting from fungal contamination of the donor cornea (2017)
  • Author(s): Kitazawa Koji、Wakimasu Koichi、Yoneda Kazuhito、Iliakis Bernie、Sotozono Chie、Kinoshita Shigeru
  • Journal Title: American Journal of Ophthalmology Case Reports Volume: 5 Pages: 103-106
  • DOI: 10.1016/j.ajoc.2016.12.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PAX6 regulates human corneal epithelium cell identity (2017)
  • Author(s): Kitazawa Koji、Hikichi Takafusa、Nakamura Takahiro、Sotozono Chie、Kinoshita Shigeru、Masui Shinji
  • Journal Title: Experimental Eye Research Volume: 154 Pages: 30-38
  • DOI: 10.1016/j.exer.2016.11.005
  • Peer Reviewed / Open Access
• [Journal Article] OVOL2 Maintains the Transcriptional Program of Human Corneal Epithelium by Suppressing Epithelial-to-Mesenchymal Transition (2016)
  • Author(s): ④K. Kitazawa, T. Hikichi, T. Nakamura, K. Mitsunaga, A. Tanaka, M. Nakamura, T. Yamakawa, S. Furukawa, M. Takasaka, N. Goshima, A. Watanabe, K. Okita, S. Kawasaki, M. Ueno, S. Kinoshita, S. Masui
  • Journal Title: Cell Reports Volume: 15 Issue: 6 Pages: 1359-1368
  • DOI: 10.1016/j.celrep.2016.04.020
  • Peer Reviewed / Open Access
• [Presentation] KLF4-depleted-human corneal epithelial cells using CRISPR/Cas9 system lost the epithelial phenotype (2018)
  • Author(s): Kitazawa K, Masuda K, Murakami R, Nakamura T, Kinoshita S, Sotozono C
  • Organizer: ARVO2018
  • Int'l Joint Research
• [Presentation] OVOL2 suppressed mesenchymal genes in human corneal epithelial cells. (2018)
  • Author(s): Kitazawa K, Hikichi T, Nakamura T, Sotozono C, Masui S, Kinoshita S
  • Organizer: Gordon Research Seminar 2018
  • Int'l Joint Research
• [Presentation] Direct Reprogramming of Human Fibroblasts into Corneal Epithelial Cells (2017)
  • Author(s): Kitazawa K
  • Organizer: Stem cells & the Eye
  • Int'l Joint Research / Invited
• [Presentation] Long-term Efficacy and Safety of Repeat Penetrating Keratoplasty: Three-Years Follow-Up. (2017)
  • Author(s): Kitazawa K, Wakimasu K, Kayukawa K, Yokota I, Inatomi T, Hieda O, Sotozono C, Kinoshita S
  • Organizer: AAO2017
  • Int'l Joint Research
• [Presentation] OVOL2 maintains a transcriptional program in human corneal epithelial cells. (2017)
  • Author(s): Kitazawa K, Hikichi T, Nakamura T, Sotozono C, Kinoshita S, Masui S
  • Organizer: ARVO2017
  • Int'l Joint Research
• [Presentation] Loss of human corneal epithelial cell identity exhibited by deletion of PAX6 using the CRISPR/Cas9 system. (2016)
  • Author(s): Kitazawa K, Hikichi T, Nakamura T, Sotozono C, Kinoshita S, Masui S
  • Organizer: ARVO 2016
  • Place of Presentation: Seattle
  • Year and Date: 2016-05-01
  • Int'l Joint Research