Chromosomal cloning of FXS-related chromosomal fragile site including pathongenic CGG triplet repeat

• Project/Area Number: 18K06183
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Tottori University
• Principal Investigator: NAKAYAMA Yuji 鳥取大学, 研究推進機構, 助教 (40432603)
• Project Period (FY): 2018-04-01 – 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 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: 脆弱X症候群 / 染色体工学 / トリプレットリピート病 / 染色体脆弱部位 / 神経変性疾患 / トリプレットリピート / 人工染色体 / ゲノム編集

Outline of Final Research Achievements

Fragile X syndrome (FXS) is most frequent familial form of intellectual disability caused by single gene, FMR1 and is categorized to trinucleotide repeat disorders (TRDs). In FXS, (CGG)n repeat witin 5'UTR of FMR1 gene becomes unstable and expand exclusively through maternal transmission. A son maternally received X chromosome with expanded (CGG)n repeats more than 200 repeats becomes FXS, as these large expansions shut down FMR1 expression. The mechanism of (CGG)n repeat expansion, a fundamental cause of FXS, has not been fully elucidated, due to lack of the appropriate experimental system(s) to recapitulate it so far. Our study demonstrated that chromosome with unstable repeat such as (CGG)n repeat can be manipulated by means of chromosome engineering and established set of rodent cells that harbors human X chromosome with various (CGG)n repeat length from FXS related individuals. Our chromosomal engineering strategy established in this study may be useful for studying other TRDs.

Academic Significance and Societal Importance of the Research Achievements

FXS罹患者は自閉症症状を併発することが多く、難治性疾患であるFXSの解明は、神経変性疾患の機能解明や治療法の開発のためにも重要である。FXSは、リピート伸長そのものが根本的病因であり、新しい治療法開拓のためにも、そのメカニズムの解明が急務である。リピートを本体とする染色体領域は、特別に染色体脆弱部位と呼ばれており、本研究はそれを機能的に独立した染色体領域としてまとめて取り扱うことが重要であるという立場に立ち、FXSのCGGリピートとその周辺領域だけをクローニングすることを目指している。本研究で活用している染色体工学技術は、将来、他のトリプレットリピート病の解析系の構築にも応用可能である。

Research Products

• [Journal Article] A transchromosomic rat model with human chromosome 21 shows robust Down syndrome features (2022)
  • Author(s): Kazuki Y, Gao FJ, Yamakawa M, Hirabayashi M, Kazuki K, Kajitani N, Miyagawa-Tomita S, Abe S, Sanbo M, Hara H, Kuniishi H, Ichisaka S, Hata Y, Koshima M, Takayama H, Takehara S, Nakayama Y, Hiratsuka M, Iida Y, Matsukura S, Noda N, Li Y, Moyer AJ, Cheng B, Singh N, Richtsmeier JT, Oshimura M, Reeves RH.
  • Journal Title: The American Journal of Human Genetics Volume: 109 Pages: 328-344
  • DOI: 10.1016/j.ajhg.2021.12.015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hsp70 promotes maturation of uromodulin mutants that cause familial juvenile hyperuricemic nephropathy and suppresses cellular damage (2022)
  • Author(s): Utami Sulistiyati Bayu、Endo Ryo、Hamada Toshihiro、Notsu Tomomi、Minato Hiroyuki、Komatsu Koji、Nakayama Yuji、Shirayoshi Yasuaki、Yamamoto Kazuhiro、Okada Shinichi、Ninomiya Haruaki、Otuki Akihiro、Hisatome Ichiro
  • Journal Title: Clinical and Experimental Nephrology Volume: Feb.25, 2022
  • DOI: 10.1007/s10157-022-02196-y
  • Peer Reviewed
• [Journal Article] Infiltration of CD204-overexpressing Macrophages Contributes to the Progression of Stage II and III Colorectal Cancer (2021)
  • Author(s): TADA YOICHIRO、MATSUMI YOSHIAKI、HARA KAZUSHI、MIYAUCHI WATARU、SUGESAWA KEN、UEJIMA CHIHIRO、TANIO AKIMITSU、KIHARA KYOICHI、YAMAMOTO MANABU、TAKANO SHUICHI、SAKAMOTO TERUHISA、NAKAYAMA YUJI、HASEGAWA TOSHIMICHI、FUJIWARA YOSHIYUKI
  • Journal Title: Anticancer Research Volume: 41 Pages: 4857-4865
  • DOI: 10.21873/anticanres.15299
  • Peer Reviewed
• [Journal Article] Construction of stable mouse artificial chromosome from native mouse chromosome 10 for generation of transchromosomic mice (2021)
  • Author(s): Abe Satoshi、Honma Kazuhisa、Okada Akane、Kazuki Kanako、Tanaka Hiroshi、Endo Takeshi、Morimoto Kayoko、Moriwaki Takashi、Hamamichi Shusei、Nakayama Yuji、Suzuki Teruhiko、Takehara Shoko、Oshimura Mitsuo、Kazuki Yasuhiro
  • Journal Title: Scientific Reports Volume: 11 Pages: 20050-20050
  • DOI: 10.1038/s41598-021-99535-y
  • Peer Reviewed
• [Journal Article] Establishment of FXS-A9 panel with a single human X chromosome from fragile X syndrome-associated individual (2021)
  • Author(s): Nakayama Yuji、Adachi Kaori、Shioda Nofirifumi、Maeta Shoya、Nanba Eiji、Kugoh Hiroyuki
  • Journal Title: Experimental Cell Research Volume: 398 Pages: 112419-112419
  • DOI: 10.1016/j.yexcr.2020.112419
  • Peer Reviewed
• [Journal Article] CGG repeat RNA G-quadruplexes interact with FMRpolyG to cause neuronal dysfunction in fragile X-related tremor/ataxia syndrome (2021)
  • Author(s): Asamitsu Sefan、Yabuki Yasushi、Ikenoshita Susumu、Kawakubo Kosuke、Kawasaki Moe、Usuki Shingo、Nakayama Yuji、Adachi Kaori、Kugoh Hiroyuki、Ishii Kazuhiro、Matsuura Tohru、Nanba Eiji、Sugiyama Hiroshi、Fukunaga Kohji、Shioda Norifumi
  • Journal Title: Science Advances Volume: 7
  • DOI: 10.1126/sciadv.abd9440
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of human immortalized mesenchymal stem cells lines for the monitoring and analysis of osteogenic differentiation in living cells (2020)
  • Author(s): Narai Takashi、Watase Ryohei、Nakayama Yuji、Kodani Isamu、Inoue Toshiaki、Kokura Kenji
  • Journal Title: Heliyon Volume: 6
  • DOI: 10.1016/j.heliyon.2020.e05398
  • Peer Reviewed / Open Access
• [Journal Article] Increased PD-1-positive macrophages in the tissue of gastric cancer are closely associated with poor prognosis in gastric cancer patients (2020)
  • Author(s): Kono, Y. Saito, H. Miyauchi, W. Shimizu, S. Murakami, Y. Shishido, Y. Miyatani, K. Matsunaga, T. Fukumoto, Y. Nakayama, Y. Sakurai, C. Hatsuzawa, K. Fujiwara, Y.
  • Journal Title: BMC Cancer Volume: 20 Pages: 175-183
  • DOI: 10.1186/s12885-020-6629-6
  • Peer Reviewed / Open Access
• [Journal Article] A novel Xist RNA-mediated chromosome inactivation model using a mouse artificial chromosome (2020)
  • Author(s): Inaoka, D. Sunamura, N. Ohira, T. Nakayama, Y. Kugoh, H.
  • Journal Title: Biotechnology Letters Volume: 42 Pages: 697-705
  • DOI: 10.1007/s10529-020-02826-z
  • Peer Reviewed / Open Access
• [Journal Article] Subclinical Epstein-Barr virus primary infection and lytic reactivation induce thyrotropin receptor autoantibodies (2019)
  • Author(s): Tamoto N, Nagata K, Hara S, Nakayama Y, Kuwamoto S, Matsushita M, Kato M, Hayashi K
  • Journal Title: Viral Immunology Volume: 32 Pages: 362-369
  • DOI: 10.1089/vim.2019.0086
  • Peer Reviewed / Open Access
• [Journal Article] Increased regulatory B cells are involved in immune evasion in patients with gastric cancer (2019)
  • Author(s): Murakami, Y. Saito, H. Shimizu, S. Kono, Y. Shishido, Y. Miyatani, K. Matsunaga, T. Fukumoto, Y. Ashida, K. Sakabe, T. Nakayama, Y. Fujiwara, Y.
  • Journal Title: Ｓｃｉｅｎｔｉｆｉｃ Ｒｅｐｏｒｔｓ Volume: 9 Pages: 13083-13083
  • DOI: 10.1038/s41598-019-49581-4
  • Peer Reviewed / Open Access
• [Journal Article] High Level Estradiol Induces EBV Reactivation and EBV gp350/220(+)CD138(+) Double-positive B Cell Population in Graves’ Disease Patients and Healthy Controls (2019)
  • Author(s): Hara S, Nagata K, Nakayama Y, Higaki K, Matsushita M, Kuwamoto S, Kato M, Hayashi K
  • Journal Title: Yonago Acta Med. Volume: 62 Issue: 2 Pages: 240-243
  • DOI: 10.33160/yam.2019.06.010
  • NAID: 130007665218
  • ISSN: 0513-5710, 1346-8049
  • Peer Reviewed / Open Access
• [Journal Article] Epstein-Barr Virus Lytic Reactivation Induces IgG4 Production by Host B Lymphocytes in Graves' Disease Patients and Controls: A Subset of Graves' Disease Is an IgG4-Related Disease-Like Condition. (2018)
  • Author(s): Nagata K, Hara S, Nakayama Y, Higaki K, Sugihara H, Kuwamoto S, Matsushita M, Kato M, Tanio S, Ishiguro K, Hayashi K.
  • Journal Title: Viral Immunol Volume: 31 (8) Pages: 540-547
  • DOI: 10.1089/vim.2018.0042
  • Peer Reviewed / Open Access
• [Presentation] Anticancer effects of hyper-coordinated germanium complex in human lung cancer cells (2019)
  • Author(s): Kashiwagi A, Nakayama Y, Ishiguro N, Nanjo M
  • Organizer: ICCOC-GTL-16
  • Int'l Joint Research
• [Presentation] Involvement of RAN translation product, FMRpolyG and neurodegeneration in Fragile X-associated Tremor/Ataxia Syndrome (FXTAS) (2019)
  • Author(s): Involvement of RAN translation product, FMRpolyG and neurodegeneration in Fragile X-associated Tremor/Ataxia Syndrome (FXTAS) : Susumu Ikenoshita1, Yuji Nakayama, Kaori Adachi, Hiroyuki Kugoh, Kazuhiro Ishii, Tohru Matsuura, Eiji Nanba, Norifumi Shioda
  • Organizer: 19th International Workshop on Fragile X and other Neurodevelopmental Disorders, Sorrento, Italy,
  • Int'l Joint Research
• [Presentation] G-quadruplex-binding metabolites rescue decreased cognitive deficits seen in FXTAS model mice (2019)
  • Author(s): Sefan Asamitsu1, Yasushi Yabuki, Yuji Nakayama, Kaori Adachi, Hiroyuki Kugoh, Hiroshi Sugiyama, Eiji Nanba, Kohji Fukuanga, Norifumi Shioda
  • Organizer: 19th International Workshop on Fragile X and other Neurodevelopmental Disorders, Sorrento, Italy,
  • Int'l Joint Research
• [Presentation] 30.Development of a novel screening system for the carcinogens in the tobacco smoke using mouse artificial chromosome (2019)
  • Author(s): Kashiwagi A, Nakayama Y, Kugoh H
  • Organizer: The 6 th Asian conference on safety and education in laboratory (Bali)
  • Int'l Joint Research
• [Presentation] Current Status of Fragile X study in Japan (2018)
  • Author(s): Yuji Nakayama, Kaori Adachi, Norifumi Shioda, Hiroyuki Kugoh and Eiji Nanba
  • Organizer: 16th International Fragile X conference
  • Int'l Joint Research
• [Presentation] 染色体操作により樹立した脆弱X症候群および関連疾患モデル細胞の解析 (2018)
  • Author(s): 中山祐二、足立香織、塩田倫史、久郷裕之、難波栄二
  • Organizer: 日本人類遺伝学会 第６３回大会
• [Presentation] 脆弱X関連振戦／失調症候群（FXTAS）におけるRAN蛋白質の病態との関与 (2018)
  • Author(s): 塩田倫史、中山祐二、足立香織、久郷裕之、難波栄二
  • Organizer: 日本人類遺伝学会 第６３回大会
• [Presentation] X染色体不活性化機構の解明に向けた人工染色体の活用 (2018)
  • Author(s): 稲岡大悟、大平崇人、押村光雄、中山祐二、久郷裕之
  • Organizer: 第41回日本分子生物学会年会