Is an eye-specific transcript revealed by comprehensive expression analysis a novel key player in corneal regeneration?

• Project/Area Number: 19K09993
• 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
• Principal Investigator: KINOSHITA Akira 長崎大学, 原爆後障害医療研究所, 准教授 (60372778)
• Co-Investigator(Kenkyū-buntansha): 小路 武彦 長崎大学, 医歯薬学総合研究科(医学系), 客員研究員 (30170179)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: Gillespie症候群 / 無虹彩 / イノシトール1,4,5三リン酸受容体I型 / 神経堤細胞 / 角膜内皮 / アクチン繊維 / フォーカルアドヒージョン / YAP / Gillespie syndrome / ITPR1 / 次世代型シーケンサー / RNA-seq / 新規転写開始点 / 前眼部組織 / 無虹彩症 / 角膜 / CRISPR/Cas9法 / 角膜再生 / ゲノム編集

Outline of Research at the Start

Gillespie症候群は無虹彩症と運動失調を特徴とする稀な遺伝病であり、ITPR1の変異で発症する。眼特異的な新規Itpr1アイソフォームが存在すること、新たに作製したItpr1ノックアウトマウスでは神経堤細胞由来の前眼部組織（角膜内皮・実質など）が消失することから、このアイソフォームが「神経堤細胞から前眼部組織の分化に関わるキープレーヤー」であると仮説を立てた。
本研究では、新規アイソフォームの眼特異的な発現調節機構、前眼部組織の形成過程における経時的なRNA発現の変化、結合タンパク質から予測される機能解明から、角膜形成過程の分子メカニズムを明らかにし、角膜再生医療の基盤研究とする。

Outline of Final Research Achievements

Gillespie syndrome (GLSP) is a rare genetic disorder characterized by cerebellar ataxia and aniridia. The GLSP-responsible gene is ITPR1 encoding a calcium channel; however, the reason for the patient with GLSP developing aniridia was unknown.
RNA-seq was performed using next-generation sequencing, and the ocular-specific novel transcript of Itpr1, which encodes an Itpr1 isoform of 218 amino acid residues, was found. This protein regulates actin fiber tension and direction and promotes nuclear translocation of the transcription coactivator YAP. This study revealed that system failure caused by GLSP mutation disrupts the differentiation of the iris and cornea tissues derived from neural crest cells, resulting in aniridia.

Academic Significance and Societal Importance of the Research Achievements

ヒトの五感の中で最も多くの情報を得ているのが視覚であり、視力障害は日常生活に大きな影響を与えQOLを著しく低下させる。そのため角膜再生とその移植治療は大きな需要がある。新たに同定した眼特異的ITPR1アイソフォームは神経堤細胞から角膜内皮および角膜ストロマの分化に必須なタンパク質をコードしていることを本研究では明らかにした。このタンパク質を利用することにより、より効率的な角膜再生法が期待できる。

Research Products

• [Journal Article] Intestinal Mucosa-Associated Lymphoid Tissue Lymphoma Transforming into Diffuse Large B-Cell Lymphoma in a Young Adult Patient with Neurofibromatosis Type 1: A Case Report (2022)
  • Author(s): Kosako Hideki、Yamashita Yusuke、Tanaka Ken、Mishima Hiroyuki、Iwamoto Ryuta、Kinoshita Akira、Murata Shin-ichi、Ohshima Koichi、Yoshiura Koh-ichiro、Sonoki Takashi、Tamura Shinobu
  • Journal Title: Medicina Volume: 58 Issue: 12 Pages: 1830-1830
  • DOI: 10.3390/medicina58121830
  • Peer Reviewed / Open Access
• [Journal Article] Hydroxychloroquine suppresses anti-GBM nephritis via inhibition of JNK/p38 MAPK signaling (2022)
  • Author(s): Torigoe Miki、Obata Yoko、Inoue Hiro、Torigoe Kenta、Kinoshita Akira、Koji Takehiko、Mukae Hiroshi、Nishino Tomoya
  • Journal Title: Clinical and Experimental Nephrology Volume: 27 Issue: 2 Pages: 110-121
  • DOI: 10.1007/s10157-022-02285-y
  • Peer Reviewed / Open Access
• [Journal Article] Identification of unique DNA methylation sites in Kabuki syndrome using whole genome bisulfite sequencing and targeted hybridization capture followed by enzymatic methylation sequencing (2022)
  • Author(s): Hamaguchi Yo、Mishima Hiroyuki、Kawai Tomoko、Saitoh Shinji、Hata Kenichiro、Kinoshita Akira、Yoshiura Koh-ichiro
  • Journal Title: Journal of Human Genetics Volume: 67 Issue: 12 Pages: 711-720
  • DOI: 10.1038/s10038-022-01083-4
  • Peer Reviewed
• [Journal Article] Alvocidib inhibits IRF4 expression via super‐enhancer suppression and adult T‐cell leukemia/lymphoma cell growth (2022)
  • Author(s): Sakamoto Hikaru、Ando Koji、Imaizumi Yoshitaka、Mishima Hiroyuki、Kinoshita Akira、Kobayashi Yuji、Kitanosono Hideaki、Kato Takeharu、Sawayama Yasushi、Sato Shinya、Hata Tomoko、Nakashima Masahiro、Yoshiura Koh‐Ichiro、Miyazaki Yasushi
  • Journal Title: Cancer Science Volume: 113 Issue: 12 Pages: 4092-4103
  • DOI: 10.1111/cas.15550
  • Peer Reviewed / Open Access
• [Journal Article] Progressive Massive Splenomegaly in an Adult Patient with Kabuki Syndrome Complicated with Immune Thrombocytopenic Purpura (2021)
  • Author(s): Mushino Toshiki、Hiroi Takayuki、Yamashita Yusuke、Suzaki Norihiko、Mishima Hiroyuki、Ueno Masaki、Kinoshita Akira、Minami Koichi、Imai Kohsuke、Yoshiura Ko-ichiro、Sonoki Takashi、Tamura Shinobu
  • Journal Title: Internal Medicine Volume: 60 Issue: 12 Pages: 1927-1933
  • DOI: 10.2169/internalmedicine.6694-20
  • NAID: 130008052635
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2021-06-15
  • Peer Reviewed / Open Access
• [Journal Article] <i>Itpr1</i> regulates the formation of anterior eye segment tissues derived from neural crest cells (2021)
  • Author(s): Kinoshita Akira、Ohyama Kaname、Tanimura Susumu、Matsuda Katsuya、Kishino Tatsuya、Negishi Yutaka、Asahina Naoko、Shiraishi Hideaki、Hosoki Kana、Tomiwa Kiyotaka、Ishihara Naoko、Mishima Hiroyuki、Mori Ryoichi、Nakashima Masahiro、Saitoh Shinji、Yoshiura Koh-ichiro
  • Journal Title: Development Volume: 148 Issue: 16 Pages: 188755-188755
  • DOI: 10.1242/dev.188755
  • Peer Reviewed / Open Access
• [Journal Article] A Patient with Kabuki Syndrome Mutation Presenting with Very Severe Aplastic Anemia (2021)
  • Author(s): Tamura Shinobu、Kosako Hideki、Furuya Yoshiaki、Yamashita Yusuke、Mushino Toshiki、Mishima Hiroyuki、Kinoshita Akira、Nishikawa Akinori、Yoshiura Ko-Ichiro、Sonoki Takashi
  • Journal Title: Acta Haematologica Volume: 145 Issue: 1 Pages: 89-96
  • DOI: 10.1159/000518227
  • Peer Reviewed
• [Journal Article] Heterozygous missense variant of the proteasome subunit β-type 9 causes neonatal-onset autoinflammation and immunodeficiency (2021)
  • Author(s): Kanazawa N, Hemmi H, Kinjo N, Ohnishi H, Hamazaki J, M.H, K.A, M.T, H.S, K.N, K.S, H.Y, I.K, N.R, T.M, Y.Y, T.S, O.T, O.T, K.T, S.I, F.Y, W.N, I.Y, K.K, O.S, T.T, N.K, M.S, Y.K, Kaisho T
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 0-0
  • DOI: 10.1038/s41467-021-27085-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comprehensive analysis for detecting radiation-specific molecules expressed during radiation-induced rat thyroid carcinogenesis. (2021)
  • Author(s): Kurohama H, Matsuda K, Kishino M, Yoshino M, Yamaguchi Y, Matsuu-Matsuyama M, Kondo H, Mitsutake N, Kinoshita A, Yoshiura KI, Nakashima M.
  • Journal Title: J Radiat Res. Volume: 62 Issue: Supplement_1 Pages: i78-i87
  • DOI: 10.1093/jrr/rraa139
  • Peer Reviewed / Open Access
• [Journal Article] Brothers with novel compound heterozygous mutations in COL27A1 causing dental and genital abnormalities (2021)
  • Author(s): Satoh Chisei、Kondoh Tatsuro、Shimizu Hitomi、Kinoshita Akira、Mishima Hiroyuki、Nishimura Gen、Miyazaki Mutsuko、Okano Kunihiko、Kumai Yoshihiko、Yoshiura Koh-ichiro
  • Journal Title: European Journal of Medical Genetics Volume: 64 Issue: 2 Pages: 104125-104125
  • DOI: 10.1016/j.ejmg.2020.104125
  • Peer Reviewed
• [Journal Article] A unique missense variant in the E1A-binding protein P400 gene is implicated in schizophrenia by whole-exome sequencing and mutant mouse models (2021)
  • Author(s): Yoshiro Morimoto, Shinji Ono, Shintaro Yoshida, Hiroyuki Mishima, Akira Kinoshita, Takeshi Tanaka, Yoshihiro Komohara, Naohiro Kurotaki, Tatsuya Kishino, Yuji Okazaki, Hiroki Ozawa, Koh-ichiro Yoshiura, Akira Imamura
  • Journal Title: Translational Psychiatry Volume: 11 Issue: 1 Pages: 132-132
  • DOI: 10.1038/s41398-021-01258-1
  • NAID: 120007000270
  • Peer Reviewed / Open Access
• [Journal Article] Targeting Adaptive IRE1α Signaling and PLK2 in Multiple Myeloma: Possible Anti-Tumor Mechanisms of KIRA8 and Nilotinib. (2020)
  • Author(s): Yamashita Y, Morita S, Hosoi H, Kobata H, Kishimoto S, Ishibashi T, Mishima H, Kinoshita A, Backes BJ, Yoshiura KI, Papa FR, Sonoki T, Tamura S
  • Journal Title: Int J Mol Sci. Volume: Aug 31;21(17) Issue: 17 Pages: 6314-6314
  • DOI: 10.3390/ijms21176314
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Expression of unfolded protein response markers in the pheochromocytoma with Waardenburg syndrome: a case report (2020)
  • Author(s): Morita Shuhei、Takeshima Ken、Ariyasu Hiroyuki、Furukawa Yasushi、Kishimoto Shohei、Tsuji Tomoya、Uraki Shinsuke、Mishima Hiroyuki、Kinoshita Akira、Takahashi Yuichi、Inaba Hidefumi、Iwakura Hiroshi、Furuta Hiroto、Nishi Masahiro、Doi Asako、Murata Shin-ichi、Yoshiura Koh-ichiro、Akamizu Takashi
  • Journal Title: BMC Endocrine Disorders Volume: 20 Issue: 1 Pages: 90-90
  • DOI: 10.1186/s12902-020-00574-9
  • Peer Reviewed / Open Access
• [Journal Article] Target Capture/Next-Generation Sequencing for Nonsyndromic Cleft Lip and Palate in the Japanese Population (2019)
  • Author(s): Shibano Masayasu、Watanabe Akira、Takano Nobuo、Mishima Hiroyuki、Kinoshita Akira、Yoshiura Koh-ichiro、Shibahara Takahiko
  • Journal Title: The Cleft Palate-Craniofacial Journal Volume: 57 Issue: 1 Pages: 80-87
  • DOI: 10.1177/1055665619857650
  • Peer Reviewed
• [Journal Article] Genome analysis of myelodysplastic syndromes among atomic bomb survivors in Nagasaki (2019)
  • Author(s): Taguchi M, Mishima H, Shiozawa Y, Hayashida C, Kinoshita A, Nannya Y, Makishima H, Horai M, Matsuo M, Sato S, Itonaga H, Kato T, Taniguchi H, Imanishi D, Imaizumi Y, Hata T, Takenaka M, Moriuchi Y, Shiraishi Y, Miyano S, Ogawa S, Yoshiura K-i, Miyazaki Y
  • Journal Title: Haematologica Volume: 105 Issue: 2 Pages: 358-365
  • DOI: 10.3324/haematol.2019.219386
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immunofluorescence analysis of DNA damage response protein p53-binding protein 1 in a case of uterine dedifferentiated leiomyosarcoma arising from leiomyoma. (2019)
  • Author(s): Matsuda K, Akazawa Y, Yamaguchi Y, Mussazhanova Z, Kurohama H, Ueki N, Kohno M, Fukushima A, Kajimura I, Hiraki H, Matsuwaki T, Kawashita S, Kinoshita A, Nakashima M
  • Journal Title: Pathology, research and practice Volume: 215 Issue: 11 Pages: 152640-152640
  • DOI: 10.1016/j.prp.2019.152640
  • Peer Reviewed / Open Access
• [Journal Article] Nonsense mutation in CFAP43 causes normal-pressure hydrocephalus with ciliary abnormalities (2019)
  • Author(s): Morimoto Y, Yoshida S, Kinoshita A, Satoh C, Mishima H, Yamaguchi N, Matsuda K, Sakaguchi M, Tanaka T, Komohara Y, Imamura A, Ozawa H, Nakashima M, Kurotaki N, Kishino T, Yoshiura K, Ono S
  • Journal Title: Neurology Volume: 92 Issue: 20
  • DOI: 10.1212/wnl.0000000000007505
  • NAID: 120006987711
  • Peer Reviewed / Open Access