ヒトｉＰＳ技術を用いた網膜色素変性症に対する新規神経保護治療の開発に向けた研究

2014 Fiscal Year Annual Research Report

• Project/Area Number: 24592647
• Research Institution: Keio University
• Principal Investigator: 小澤 洋子 慶應義塾大学, 医学部, 講師 (90265885)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 網膜 / 幹細胞 / iPS / 再生 / 神経保護

Outline of Annual Research Achievements

網膜色素変性症は、遺伝子異常を原因とし、徐々に進行する疾患で、失明原因の上位を占める。早期に診断が付いても進行を抑制する安定した治療法はないのが現状である。異常遺伝子が明らかになっていても、網膜神経細胞障害（死）のメカニズムは今のところ、わかっていないものが多いためである。その理由の一つが、患者網膜を採取して、培養したり解析したりすることが困難であることである。そこで、患者の異常遺伝子を持つ人工多能性細胞(induced-pluripotent stem cell; iPS細胞)を用いて病態メカニズムを解析し、新規の神経保護治療法の開発に向けた研究を開始した。
具体的には、網膜色素変性症患者の皮膚細胞由来のiPS細胞を樹立し、これを網膜細胞に分化誘導した。また、コントロールiPS細胞からも網膜細胞を分化誘導した。これらの網膜細胞の中で視細胞だけに注目して解析を行うために、視細胞特異的に発現するNRL遺伝子のプロモーター支配下にGFPを発現するベクターを、分化誘導中の細胞にウイルスを用いて感染させ標識し、フローサイトメトリーを用いて選択的に回収した。生存細胞を測定するとともに、回収した細胞のアポトーシスマーカー、小胞体ストレスマーカーを比較解析したところ、患者由来細胞では、生存細胞が少なく、アポトーシスマーカー、小胞体ストレスマーカーが亢進していた。また、回収した視細胞の培養にラパマイシンを投与すると、生存細胞は増加し、アポトーシスマーカー、小胞体ストレスマーカーの発現が抑制されるという結果を得た。

Research Products

• [Journal Article] Clinical and Molecular Characteristics of Childhood-onset Stargardt Disease. (2015)
  • Author(s): Fujinami K, Jana Zernant, Ravinder K Chana, Genevieve A Wright, Tsunoda K, Ozawa Y, Tsubota K, Anthony G Robson, Graham E Holder, Rando Allikmets, Michel Michaelides, FACS; Anthony T Moore.
  • Journal Title: Ophthalmology. Volume: 122 Pages: 326-334
  • DOI: 10.1016/j.ophtha.2014.08.012.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Wickerhamomyces anomalus Fungal Keratitis Responds to Topical Treatment with Antifungal Micafungin. (2015)
  • Author(s): Kamoshita M, Matsumoto Y, Nishimura K, Katono Y, Murata M, Ozawa Y, Shimmura S, Tsubota K.
  • Journal Title: Journal of Infection and Chemotherapy. Volume: 21 Pages: 141-143
  • DOI: 10.1016/j.jiac.2014.08.019.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Phase II enzyme induction by a carotenoid, lutein, in a PC12D neuronal cell line. (2014)
  • Author(s): Miyake S, Kobayashi S, Tsubota K, Ozawa Y
  • Journal Title: Biochem Biophys Res Commun Volume: 446 Pages: 535-540
  • DOI: 10.1016/j.bbrc.2014.02.135
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Angiotensin II type 1 receptor blockade suppresses light-induced neural damage in the mouse retina. (2014)
  • Author(s): Narimatsu T, Ozawa Y, Miyake S, Nagai N, Tsubota K.
  • Journal Title: Free Radical Biol Med Volume: 71 Pages: 176-185
  • DOI: 10.1016/j.freeradbiomed.2014.03.020.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Predictive factors for non-responders to intravitreal ranibizumab treatment in age-related macular degeneration. (2014)
  • Author(s): Suzuki M, Nagai N, Izumi-Nagai K, Shinoda H, Koto T, Uchida A, Mochimaru H, Yuki K, Sasaki M, Tsubota K, Ozawa Y.
  • Journal Title: Br J Ophthalmoｌ Volume: 98 Pages: 1186-1191
  • DOI: 10.1136/bjophthalmol-2013-304670.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Association of Maternal Age to Development and Progression of Retinopathy of Prematurity in Infants of Gestational Age under 33 Weeks. (2014)
  • Author(s): Uchida A, Miwa M, Shinoda H, Koto T, Nagai N, Mochimaru H, Tomita Y, Sasaki M, Ikeda K, Tsubota K, Ozawa Y
  • Journal Title: J Ophthalmol. Volume: 2014 Pages: 187929-187929
  • DOI: 10.1155/2014/187929.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Early Signs of Exudative Age-related Macular Degeneration in Asians (2014)
  • Author(s): Sasaki M, Kawasaki R, Uchida A, Koto T, Shinoda H, Tsubota K, Tien Yin Wong, Ozawa Y
  • Journal Title: Optometry and Vision Science. Volume: 91 Pages: 849-853
  • DOI: 10.1097/OPX.0000000000000317.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Resveratrol Prevents the Development of Choroidal Neovascularization by Modulating AMP-activated Protein Kinase in Macrophages and Other Cell Types. (2014)
  • Author(s): Nagai N, Kubota S, Tsubota K, Ozawa Y.
  • Journal Title: J Nutr Biochem. Volume: 25 Pages: 1218-1225
  • DOI: 10.1016/j.jnutbio.2014.05.015.
  • Peer Reviewed / Open Access
• [Journal Article] The use of induced pluripotent stem cells to reveal pathogenic gene mutations and explore treatments for retinitis pigmentosa. (2014)
  • Author(s): Yoshida T, Ozawa Y, Suzuki K, Yuki K, Ohyama M, Akamatsu W, Matsuzaki Y, Shimmura S, Mitani K, Tsubota K, Okano H.
  • Journal Title: Mol Brain. Volume: 7 Pages: 45-45
  • DOI: 10.1186/1756-6606-7-45.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Decreased proteasomal activity causes photoreceptor degeneration in mice. (2014)
  • Author(s): Ando R, Noda K, Tomaru U, Kamoshita M, Ozawa Y, Notomi S, Hisatomi T, Noda M, Kanda A, Ishibashi T, Kasahara M, Ishida S.
  • Journal Title: Invest Ophthalmol Vis Sci. Volume: 55 Pages: 4682-4690
  • DOI: 10.1167/iovs.13-13272.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] AMPK-NF-κB Axis in the Photoreceptor Disorder during Retinal Inflammation. (2014)
  • Author(s): Kamoshita M, Ozawa Y, Kubota S, Miyake S, Tsuda C, Nagai N, Yuki K, Shimmura S, Umezawa K, Tsubota K.
  • Journal Title: PLOS ONE Volume: 9 Pages: 103013-103013
  • DOI: 10.1371/journal.pone.0103013.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Intraoperative and fluorescein angiographic findings of a secondary macular hole associated with age-related macular degeneration treated by pars plana vitrectomy. (2014)
  • Author(s): Okamoto T, Shinoda H, Kurihara T, Nagai N Tsubota K, Ozawa Y.
  • Journal Title: BMC Ophthalmology. Volume: 14 Pages: 114-114
  • DOI: 10.1186/1471-2415-14-114.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Management of AMD; anti-VEGF and anti-oxidant approaches. (2014)
  • Author(s): Ozawa Y
  • Organizer: Autumn meeting of Taiwan Ophthalmology Academy
  • Place of Presentation: Chiayi, Taiwan
  • Year and Date: 2014-10-05 – 2014-10-05
  • Invited
• [Presentation] Anti-VEGF therapy in DME (2014)
  • Author(s): Ozawa Y
  • Organizer: Autumn meeting of Taiwan Ophthalmology Academy
  • Place of Presentation: Chiayi, Taiwan
  • Year and Date: 2014-10-05 – 2014-10-05
  • Invited
• [Presentation] Oxidative stress in the retina and the pathogenesis (2014)
  • Author(s): Ozawa Y
  • Organizer: Inflammatory mechanisms/oxidative stress
  • Place of Presentation: ISOHK-APGC, Hong Kong,
  • Year and Date: 2014-09-26 – 2014-09-26
  • Invited
• [Presentation] Circadian Regulation of Sirtuins in the Eye. Education Course. (2014)
  • Author(s): Ozawa Y
  • Organizer: The Association for Research in Vision and Ophthalmology
  • Place of Presentation: Orland, FL, USA
  • Year and Date: 2014-05-04 – 2014-05-08
  • Invited
• [Presentation] Management of ME secondary to CRVO Macular edema secondary to Central Retinal Vein Occlusion. CRVO. (2014)
  • Author(s): Ozawa Y
  • Organizer: The Association for Research in Vision and Ophthalmology
  • Place of Presentation: Orland, FL, USA
  • Year and Date: 2014-05-04 – 2014-05-08
  • Invited