Functional role of truncated dystrophin with deletion exon 45-55

• Project/Area Number: 25860306
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Experimental pathology
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: TANIHATA Jun 独立行政法人国立精神・神経医療研究センター, 神経研究所 遺伝子疾患治療研究部, 流動研究員 (00508426)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 筋ジストロフィー / 筋小胞体 / SERCA / ｎＮＯＳ / カルシウム動態 / 細胞内カルシウム動態 / 短縮型ジストロフィン / nNOS / 細胞内カルシウム濃度 / リアノジン受容体

Outline of Final Research Achievements

Duchenne muscular dystrophy (DMD) is caused by lack of dystrophin. Exon skipping is a promising treatment for DMD and “exon 45-55 skipping” strategy is though to be one of the goals of this therapy, partly because Becker muscular dystrophy patients with exon 45-55 in-frame deletion show very mild skeletal muscle symptoms. However the function of exon 45-55 deleted dystrophin is not well investigated. We generated Tg/mdx mice and extensively analyzed their phenotypes. The improvement of muscle pathology and muscle function of Tg/mdx were confirmed. On the other hand, nNOS of Tg/mdx exists as an activated form and its localization was changed from sarcolemma to cytosol. We found that activated nNOS in Tg/mdx led to hyper-nitrosylation of the RyR1 and subsequent constant Ca2+ release from SR to cytosol. These phenomena were also confirmed in mdx. However, the function of RyR1 stimulated by caffeine and SERCA were maintained in Tg/mdx and wild type but not in mdx.

Research Products

• [Journal Article] Low intensity training of mdx mice reduces carbonylation and increases expression levels of proteins involved in energy metabolism and muscle contraction. (2015)
  • Author(s): Hyzewicz J, Tanihata J, Kuraoka M, Ito N, Miyagoe-Suzuki Y, Takeda S
  • Journal Title: Free Radic Biol Med Volume: 82 Pages: 122-136
  • Peer Reviewed / Open Access
• [Journal Article] Long-term efficacy of systemic multiexon skipping targeting dystrophin exons 45-55 with a cocktail of vivo-morpholinos in mdx52 mice. (2015)
  • Author(s): Echigoya Y, Aoki Y, Miskew B, Panesar D, Touznik A, Nagata T, Tanihata J, Nakamura A, Nagaraju K, Yokota T
  • Journal Title: Mol Ther Nucleic Acids Volume: 4 Pages: e225-e225
  • DOI: 10.1038/mtna.2014.76
  • Peer Reviewed / Open Access
• [Journal Article] Enhancement of exon skipping in mdx52 mice by 2′-O-methyl-2-thioribothymidine incorporation into phosphorothioate oligonucleotides. (2015)
  • Author(s): Masaki Y, Inde T, Nagata T, Tanihata J, Kanamori T, Seio K, Takeda S, Sekine M
  • Journal Title: Med Chem Commun Volume: 6 Issue: 6 Pages: 630-633
  • DOI: 10.1039/c5md00079c
  • Peer Reviewed / Open Access
• [Journal Article] Nakamura K, Fujii W, Tsuboi M, Tanihata J, Teramoto N, Takeuchi S, Naito K, Yamanouchi K, Nishihara M (2014)
  • Author(s): Nakamura K, Fujii W, Tsuboi M, Tanihata J, Teramoto N, Takeuchi S, Naito K, Yamanouchi K, Nishihara M
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 5635-5635
  • DOI: 10.1038/srep05635
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Mutation types and aging differently affect revertant fiber expansion in dystrophic mdx and mdx52 mice. (2013)
  • Author(s): Echigoya Y, Lee J, Rodrigues M, Nagata T, Tanihata J, Nozohourmehrabad A, Panesar D, Miskew B, Aoki Y, Yokota T.
  • Journal Title: PLoS One Volume: 8(7) Issue: 7 Pages: e69194-e69194
  • DOI: 10.1371/journal.pone.0069194
  • Peer Reviewed
• [Presentation] Truncated dystrophin with exon 45-55 deletion induced muscle atrophy and fiber type change through the hyper-nitrosylation of the ryanodine receptor type-1 and constant release of Ca2+ to the cytosol. (2014)
  • Author(s): Tanihata J, Nagata T, Saito T, Ito N, Aoki Y, Nakamura A, Takeda S
  • Organizer: 19th International Congress of the World Muscle Society
  • Place of Presentation: Academic center of Berlin, Berlin, Germany
  • Year and Date: 2014-10-07 – 2014-10-11
• [Presentation] Exon45-55を欠失した短縮型dystrophinはRyR1をニトロシル化し、細胞内Ca2＋濃度を上昇させる． (2014)
  • Author(s): 谷端 淳，永田哲也，齊藤 崇，伊藤尚基，中村昭則，武田伸一
  • Organizer: 第69回日本体力医学会大会
  • Place of Presentation: 長崎・長崎大学
  • Year and Date: 2014-09-19 – 2014-09-21
• [Presentation] Proteomic analysis shows that low intensity training reduces the carbonylation level and increases the expression of energy metabolism and muscle contraction proteins in mdx skeletal muscle. (2014)
  • Author(s): Hyzewicz J, Tanihata J, Kuraoka M, Ito N, Miyagoe-Suzuki Y, Takeda S
  • Organizer: 43th European Muscle Conference
  • Place of Presentation: University of Salzburg, Salzburg, Austria
  • Year and Date: 2014-09-10 – 2014-09-14
• [Presentation] CRISPR/Cas法を用いた筋ジストロフィーモデルラットの作出 (2014)
  • Author(s): 中村克行，藤井 渉，坪井誠也，谷端 淳，寺本奈保美，竹内志帆，内藤邦彦，山内啓太郎，西原真杉
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 札幌・北海道大学
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] Generation of Dystrophin mutated rats with a CRISPR/Cas system as a new animal model of muscular dystrophy. (2014)
  • Author(s): Katsuyuki Nakamura, Wataru Fujii, Masaya Tsuboi, Jun Tanihata, Naomi Teramoto, Shiho Takeuchi, Kunihiko Naito, Keitaro Yamanouchi, Masugi Nishihara
  • Organizer: 2014 FASEB Science Research Conferences（Skeletal Muscle Satellite and Stem Cells）
  • Place of Presentation: Steamboat Springs, Colorado, USA
  • Year and Date: 2014-07-20 – 2014-07-25
• [Presentation] Low intensity endurance training restores the expression of troponin T and myosin binding protein C Fast type isoforms in gastrocnemius of mdx mice. (2014)
  • Author(s): Janek Hyzewicz，谷端 淳，倉岡睦季，伊藤尚基，鈴木友子，武田伸一
  • Organizer: 第35回日本炎症・再生医学会
  • Place of Presentation: 沖縄・万国津梁館
  • Year and Date: 2014-07-01 – 2014-07-04
• [Presentation] Truncated dystrophin with exon 45-55 deletion induced muscle atrophy and fiber type change through the hyper-nitrosylation of the ryanodine receptor type-1 and constant release of Ca2+ to the cytosol (2014)
  • Author(s): Jun Tanihata, Tetsuya Nagata, Takashi Saito, Naoki Ito, Yoshitsugu Aoki, Akinori Nakamura, Shin’ichi Takeda
  • Organizer: 19th International WMS Congress
  • Place of Presentation: Langenbeck-Virchow-Haus, Ｂｅｒｌｉｎ， Ｇｅｒｍａｎｙ
• [Presentation] Exon45-55を欠失した短縮型dystrophinの機能的役割の解明
  • Author(s): 谷端 淳、永田哲也、齋藤 崇、青木吉嗣、中村昭則、武田伸一
  • Organizer: 第68回日本体力医学会
  • Place of Presentation: 日本教育会館
• [Patent(Industrial Property Rights)] 筋細胞損傷抑制剤 (2014)
  • Inventor(s): 橋戸和夫, 松坂恭成, 谷端 淳
  • Industrial Property Rights Holder: 橋戸和夫, 松坂恭成, 谷端 淳
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-226518
  • Filing Date: 2014-11-06