Comprehensive genetic screening and establishment of novel therapeutic method in retinitis pigmentosa

• Project/Area Number: 19K09929
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Nagasaki University (2020-2021); Kyoto University (2019)
• Principal Investigator: Oishi Akio 長崎大学, 医歯薬学総合研究科(医学系), 准教授 (50572955)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 網膜色素変性 / 遺伝子検査 / 遺伝型表現型解析 / 環境因子 / 分化誘導 / stop codon readthrough / 次世代シーケンサー / 遺伝子治療

Outline of Research at the Start

網膜色素変性は遺伝性疾患であり、進行性の網膜障害を来す。本研究では研究代表者がこれまで行ってきた網羅的遺伝子診断の成果を軸とし、premature termination codonの変異を持つ患者から樹立した疾患特異iPS細胞を用いてstop codon readthroughによる治療法を開発することを第一の目的とする。さらに既報の遺伝子を網羅的に調べても原因遺伝子が特定できなかった症例から、家系解析および人工知能を用いて原因となる遺伝子の候補を絞り込み、動物実験で表現型を確認することで新規遺伝子の同定を目指す。

Outline of Final Research Achievements

We performed comprehensive genetic screening in Japanese patients with retinitis pigmentosa. We established iPS cells from patients harboring specific variants. We identified causative variants in 98 patients and confirmed that EYS is the major cause of retinitis pigmentosa in Japan. We also reported genotype and phenotype of patients with some specific genes including CYP4V2, PROM1, and PRPH2. We found that environmental factors such as tobacco and light exposure may affect retinal degeneration. We established a new method to generate photoreceptor like cells from iPS cells, which consists of induction of CRX and NEUROD1.

Academic Significance and Societal Importance of the Research Achievements

網膜色素変性には多くの原因遺伝子があり、その頻度には人種間での差も大きい。日本で特に多い遺伝子、およびバリアントを特定することは、今後治療法を考えるうえでより多くの方を対象と出来るようにするための基礎的な情報となる。また現時点で確立した治療方法のないこの疾患について、喫煙が網膜変性を進行させる可能性を示したことは、禁煙というすぐにでも実行可能な介入を勧める根拠となる。またiPS細胞から効率的に視細胞様の細胞を分化させる方法は今後iPS細胞を使った研究を行う上で広く応用可能な技術となる。

Research Products

• [Journal Article] One-step induction of photoreceptor-like cells from human iPSCs by delivering transcription factors (2022)
  • Author(s): Otsuka Yuki、Imamura Keiko、Oishi Akio、Kondo Takayuki、Suga Mika、Yada Yuichiro、Shibukawa Ran、Okanishi Yasue、Sagara Yukako、Tsukita Kayoko、Tsujikawa Akitaka、Inoue Haruhisa
  • Journal Title: iScience Volume: 25 Issue: 4 Pages: 103987-103987
  • DOI: 10.1016/j.isci.2022.103987
  • Peer Reviewed / Open Access
• [Journal Article] Association between the number of visual fields and the accuracy of future prediction in eyes with retinitis pigmentosa (2021)
  • Author(s): Asaoka Ryo、Oishi Akio、Fujino Yuri、Murata Hiroshi、Azuma Keiko、Miyata Manabu、Obata Ryo、Inoue Tatsuya
  • Journal Title: BMJ Open Ophthalmology Volume: 6 Issue: 1 Pages: e000900-e000900
  • DOI: 10.1136/bmjophth-2021-000900
  • Peer Reviewed
• [Journal Article] Genetic and Phenotypic Landscape of PRPH2-Associated Retinal Dystrophy in Japan (2021)
  • Author(s): Oishi Akio、Fujinami Kaoru、Mawatari Go、Naoi Nobuhisa、Ikeda Yasuhiro、Ueno Shinji、Kuniyoshi Kazuki、Hayashi Takaaki、Kondo Hiroyuki、Mizota Atsushi、Shinoda Kei、Kusuhara Sentaro、Nakamura Makoto、Iwata Takeshi、Tsujikawa Akitaka、Tsunoda Kazushige
  • Journal Title: Genes Volume: 12 Issue: 11 Pages: 1817-1817
  • DOI: 10.3390/genes12111817
  • Peer Reviewed / Open Access
• [Journal Article] Detecting Progression of Retinitis Pigmentosa Using the Binomial Pointwise Linear Regression Method (2021)
  • Author(s): Asano Shotaro、Oishi Akio、Asaoka Ryo、Fujino Yuri、Murata Hiroshi、Azuma Keiko、Miyata Manabu、Obata Ryo、Inoue Tatsuya
  • Journal Title: Translational Vision Science & Technology Volume: 10 Issue: 13 Pages: 15-15
  • DOI: 10.1167/tvst.10.13.15
  • Peer Reviewed
• [Journal Article] Detection Sensitivity of Retinitis Pigmentosa Progression Using Static Perimetry and Optical Coherence Tomography (2021)
  • Author(s): Hasegawa Tomoko、Oishi Akio、Ikeda Hanako Ohashi、Numa Shogo、Miyata Manabu、Otsuka Yuki、Oishi Maho、Tsujikawa Akitaka
  • Journal Title: Translational Vision Science & Technology Volume: 10 Issue: 8 Pages: 31-31
  • DOI: 10.1167/tvst.10.8.31
  • Peer Reviewed / Open Access
• [Journal Article] Evaluation of outer nuclear layer overshadowed by retinal vessels in retinitis pigmentosa (2021)
  • Author(s): Otsuka Yuki、Oishi Akio、Miyata Manabu、Uji Akihito、Oishi Maho、Hasegawa Tomoko、Numa Shogo、Ikeda Hanako Ohashi、Tsujikawa Akitaka
  • Journal Title: Eye Volume: - Issue: 5 Pages: 1042-1049
  • DOI: 10.1038/s41433-021-01578-y
  • Peer Reviewed
• [Journal Article] Development and validation of a visual field cluster in retinitis pigmentosa (2021)
  • Author(s): Omoto Takashi、Oishi Akio、Asaoka Ryo、Fujino Yuri、Murata Hiroshi、Azuma Keiko、Miyata Manabu、Obata Ryo、Inoue Tatsuya
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 9671-9671
  • DOI: 10.1038/s41598-021-89233-0
  • Peer Reviewed / Open Access
• [Journal Article] Clinical Characteristics, Differential Diagnosis and Genetic Analysis of Concentric Retinitis Pigmentosa (2021)
  • Author(s): Nakahara Mei、Oishi Akio、Miyata Manabu、Ikeda Hanako Ohashi、Hasegawa Tomoko、Numa Shogo、Otsuka Yuki、Oishi Maho、Matsuda Fumihiko、Tsujikawa Akitaka
  • Journal Title: Life Volume: 11 Issue: 3 Pages: 260-260
  • DOI: 10.3390/life11030260
  • Peer Reviewed / Open Access
• [Journal Article] Genotype and Long-term Clinical Course of Bietti Crystalline Dystrophy in Korean and Japanese Patients. (2021)
  • Author(s): Murakami Y, Koyanagi Y, Fukushima M, Yoshimura M, Fujiwara K, Akiyama M, Momozawa Y, Ueno S, Terasaki H, Oishi A, Miyata M, Ikeda H, Tsujikawa A, Mizobuchi K, Hayashi T, Fujinami K, Tsunoda K, Park JY, Han J, Kim M, Lee CS, Kim SJ, Park TK, Joo K, Woo SJ, Ikeda Y, Sonoda KH.
  • Journal Title: Ophthalmol Retina Volume: - Issue: 12 Pages: 1269-1279
  • DOI: 10.1016/j.oret.2021.02.009
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Generation of a human induced pluripotent stem cell line, BRCi005-A, derived from a Best disease patient with BEST1 mutations (2020)
  • Author(s): Kamata Kazuma、Otsuka Yuki、Imamura Keiko、Oishi Akio、Kondo Takayuki、Suga Mika、Shibukawa Ran、Okanishi Yasue、Sagara Yukako、Tsukita Kayko、Yasukawa Tsutomu、Usui Hideaki、Muguruma Keiko、Tsujikawa Akitaka、Inoue Haruhisa
  • Journal Title: Stem Cell Research Volume: 45 Pages: 101782-101782
  • DOI: 10.1016/j.scr.2020.101782
  • Peer Reviewed / Open Access
• [Journal Article] Generation of a human induced pluripotent stem cell line, BRCi004-A, derived from a patient with age-related macular degeneration (2020)
  • Author(s): Kamata Kazuma、Otsuka Yuki、Imamura Keiko、Oishi Akio、Kondo Takayuki、Suga Mika、Shibukawa Ran、Okanishi Yasue、Sagara Yukako、Tsukita Kayko、Yasukawa Tsutomu、Usui Hideaki、Muguruma Keiko、Tsujikawa Akitaka、Inoue Haruhisa
  • Journal Title: Stem Cell Research Volume: 45 Pages: 101787-101787
  • DOI: 10.1016/j.scr.2020.101787
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and genetic characteristics of 10 Japanese patients with PROM1-associated retinal disorder: A report of the phenotype spectrum and a literature review in the Japanese population. (2020)
  • Author(s): Fujinami K, Oishi A, Yang L, Arno G, Pontikos N, Yoshitake K, Fujinami-Yokokawa Y, Liu X, Hayashi T, Katagiri S, Mizobuchi K, Mizota A, Shinoda K, Nakamura N, Kurihara T, Tsubota K, Miyake Y, Iwata T, Tsujikawa A, Tsunoda K; Japan Eye Genetics Consortium study group.
  • Journal Title: Am J Med Genet C Semin Med Genet. Volume: 184 Issue: 3 Pages: 656-674
  • DOI: 10.1002/ajmg.c.31826
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Wavelength of light and photophobia in inherited retinal dystrophy (2020)
  • Author(s): Otsuka Yuki、Oishi Akio、Miyata Manabu、Oishi Maho、Hasegawa Tomoko、Numa Shogo、Ikeda Hanako Ohashi、Tsujikawa Akitaka
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 14798-14798
  • DOI: 10.1038/s41598-020-71707-2
• [Journal Article] Effect of smoking on macular function and retinal structure in retinitis pigmentosa (2020)
  • Author(s): Oishi Akio、Noda Kazunori、Birtel Johannes、Miyake Masahiro、Sato Atsuyasu、Hasegawa Tomoko、Miyata Manabu、Numa Shogo、Charbel Issa Peter、Tsujikawa Akitaka
  • Journal Title: Brain Communications Volume: 2 Issue: 2
  • DOI: 10.1093/braincomms/fcaa117
• [Presentation] 網膜ジストロフィの原因遺伝子と環境因子 (2020)
  • Author(s): 大石明生
  • Organizer: 長崎県眼科医会学術講演会
  • Invited
• [Presentation] 網膜ジストロフィの遺伝子と環境因子 (2019)
  • Author(s): 大石明生
  • Organizer: 京大眼科同窓会学会
  • Invited
• [Presentation] 網膜血管陰影を利用した網膜色素変性患者に対する光障害の影響の検討 (2019)
  • Author(s): 大塚悠生, 大石明生, 宮田学, 宇治彰人, 大石真秀, 長谷川智子, 沼尚吾, 辻川明孝
  • Organizer: 日本網膜硝子体学会
• [Presentation] Investigating the origin of photophobia in inherited retinal dystrophy (2019)
  • Author(s): Otsuka Y, Oishi A, Miyata M, Oishi M, Hasegawa T, Numa S, Tsujikawa A
  • Organizer: EURETINA
  • Int'l Joint Research