Elucidation of the molecular mechanism and development of therapeutic method of exon skip caused by mutation

• Project/Area Number: 18K07873
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kanagawa Children's Medical Center (Clinical Research Institute) (2021); Tokyo Medical and Dental University (2018-2020)
• Principal Investigator: NARUTO TAKUYA 地方独立行政法人神奈川県立病院機構神奈川県立こども医療センター(臨床研究所), 臨床研究所, 主任研究員 (60438124)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: スプライシング変異 / スプライス変異 / イントロン変異

Outline of Final Research Achievements

Abnormal RNA splicing is the cause of the disease due to mutations in the cis-regulatory sequence of RNA splicing and changes in trans-regulatory factors. RNA-Seq was performed from the patient's blood verified splicing patterns and expression levels related to splicing mutations in actual patients using StringTie. A minigene assay was used to verify whether mutations predicted by splicing prediction software had occurred. These results showed that the exon-terminal deletion was transcribed into RNA using contiguous intron sequences, strongly preserving the original splicing sites and not using the predicted splicing sites in introns.

Academic Significance and Societal Importance of the Research Achievements

エクソン欠失の報告が数少ない遺伝子におけるスプライシング変異の詳細な解析を行い、ｍRNAのパターンと発現の解析をした。数多くのスプライシング予測ソフトが公開されているが、本研究の結果を組み入れることによりDNA変異から生じるRNAの予測向上の一助となることが期待される。また、迅速な結果解釈による病気の早期診断や治療の向上が期待される。

Research Products

• [Journal Article] Update of the genotype and phenotype of KMT2D and KDM6A by genetic screening of 100 patients with clinically suspected Kabuki syndrome (2020)
  • Author(s): Murakami H, Tsurusaki Y, Enomoto K, Kuroda Y, Yokoi T, Furuya N, Yoshihashi H, Minatogawa M, Abe-Hatano C, Ohashi I, Nishimura N, Kumaki T, Enomoto Y, Naruto T, Iwasaki F, Harada N, Ishikawa A, Kawame H, Sameshima K, Yamaguchi Y, Kobayashi M, Tominaga M, Kuroki Y, Kurosawa K. et al.
  • Journal Title: American Journal of Medical Genetics Part A Volume: 182 Issue: 10 Pages: 2333-2344
  • DOI: 10.1002/ajmg.a.61793
  • Peer Reviewed
• [Journal Article] A deep intronic mutation of c.1166-285?T?>?G in SLC46A1 is shared by four unrelated Japanese patients with hereditary folate malabsorption (HFM) (2019)
  • Author(s): Tozawa Yusuke、Abdrabou Shimaa Said Mohamed Ali、Nogawa-Chida Natsuko、Nishiuchi Ritsuo、Ishida Toshiaki、Suzuki Yuichi、Sano Hideki、Kobayashi Ryoji、Kishimoto Kenji、Ohara Osamu、Imai Kohsuke、Naruto Takuya、Kobayashi Kunihiko、Ariga Tadashi、Yamada Masafumi
  • Journal Title: Clinical Immunology Volume: 208 Pages: 108256-108256
  • DOI: 10.1016/j.clim.2019.108256
  • Peer Reviewed / Open Access / Int'l Joint Research