Research of signal transduction system in antisense therapy for muscular dystrophy

• Project/Area Number: 20591223
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kobe University
• Principal Investigator: TAKESHIMA Yasuhiro Kobe University, 大学院・医学研究科, 特命教授 (40281141)
• Co-Investigator(Kenkyū-buntansha): MATSUO Masafumi 神戸大学, 医学研究科, 教授 (10157266) ; YAGI Mariko 神戸大学, 医学研究科, 助教 (60362787)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2009: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2008: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: アンチセンスオリゴヌクレオチド / 筋ジストロフィー / シグナル伝達因子 / エクソンスキッピング / 筋ジストロフー / NF-κB

Research Abstract

We have shown the clinical effectiveness of antisense therapy for Duchenne muscular dystrophy (DMD) which converts out-of-frame to in-frame mutation by inducing the exon skipping by antisense oligonucleotide. To enhance the effect of this therapy, we developed the in vitro model system of antisense therapy based on the mutation spectrum of Japanese DMD cases. Furthermore, the effect of antisense oligonucleotide on dystrophin production and signal transduction system was investigated. The effect of this therapy was different among DMD cases, and it is important to examine the effect of antisense therapy using individual in vitro system of each case.

Research Products

• [Journal Article] Molecular characterization of the 5'-UTR of retinal dystrophin reveals a cryptic intron that regulates translational activity. (2010)
  • Author(s): Kubokawa I, Takeshima Y, Ota M, Enomoto M, Okizuka Y, Mori T, Nishimura N, Awano H, Yagi M, Matsuo M.
  • Journal Title: Mol Vis. 16 Pages: 2590-2597
• [Journal Article] Contemporary retrotransposition of a novel non-coding gene induces exon-skipping in dystrophin mRNA. (2010)
  • Author(s): Awano H, Malueka RG, Yagi M, Okizuka Y, Takeshima Y, Matsuo M.
  • Journal Title: J Hum Genet. 55 Pages: 785-790
  • NAID: 10030737817
• [Journal Article] Mutation spectrum of the dystrophin gene in 442 Duchenne/Becker muscular dystrophy cases from one Japanese referral center. (2010)
  • Author(s): Takeshima Y, Yagi M, Okizuka Y, Awano H, Zhang Z, Yamauchi Y, Nishio H, Matsuo M.
  • Journal Title: J Hum Genet. 55 Pages: 379-388
  • NAID: 10030735518
• [Journal Article] Low incidence of limb-girdle muscular dystrophy type 2C revealed by a mutation study in Japanese patients clinically diagnosed with DMD. (2010)
  • Author(s): Okizuka Y, Takeshima Y, Itoh K, Zhang Z, Awano H, MarMarrar, Mar, Marma K, Kumagai T, Yagi M, Matsuo M.
  • Journal Title: BMC Med Genet. 30
• [Journal Article] Mutation spectrum of the dystrophin gene in 442 Duchenne/Becker muscular dystrophy cases from one Japanese referral center. (2010)
  • Author(s): Takeshima Y
  • Journal Title: Journal of Human Genetics. Volume: 55 Pages: 379-388
  • NAID: 10030735518
  • Peer Reviewed
• [Journal Article] Low incidence of limb-girdle muscular dystrophy type 2C revealed by a mutation study in Japanese patients clinically diagnosed with DMD. (2010)
  • Author(s): Okizuka Y
  • Journal Title: BMC Medical Genetics. Volume: 11 Pages: 49-49
  • Peer Reviewed
• [Journal Article] A Japanese child with asymptomatic elevation of serum creatine kinase shows PTRF-CAVIN mutation matching with congenital generalized lipodystrophy type 4. (2010)
  • Author(s): Dwianingsih EK
  • Journal Title: Molecular Genetics and Metabolism. Volume: 101 Pages: 233-237
  • Peer Reviewed
• [Journal Article] Insertion of the IL1RAPL1 gene into the duplication junction of the dystrophin gene. (2009)
  • Author(s): Zhang Z, Yagi M, Okizuka Y, Awano H, Takeshima Y, Matsuo M.
  • Journal Title: J Hum Genet. 54 Pages: 466-473
  • NAID: 10030731391
• [Journal Article] Small mutations detected by multiplex ligation-dependent probe amplification of the dystrophin gene. (2009)
  • Author(s): Okizuka Y, Takeshima Y, Awano H, Zhang Z, Yagi M, Matsuo M.
  • Journal Title: Genet Test Mol Biomarkers. 13 Pages: 427-431
• [Journal Article] In vitro splicing analysis showed that availability of a cryptic splice site is not a determinant for alternative splicing patterns caused by +1G-->A mutations in introns of the dystrophin gene. (2009)
  • Author(s): Habara Y, Takeshima Y, Awano H, Okizuka Y, Zhang Z, Saiki K, Yagi M, Matsuo M.
  • Journal Title: J Med Genet. 46 Pages: 542-547
• [Journal Article] In vitro splicing analysis showed that availability of a cryptic splice site is not a determinant for alternative splicing patterns caused by +1G→A mutations in introns of the dystronhin gene (2009)
  • Author(s): Habara Y
  • Journal Title: Journal of Medical Genetics 46 Pages: 542-547
  • Peer Reviewed
• [Journal Article] Small mutations detected by multiplex ligation-dependent probe amplification of the dystrophin gene (2009)
  • Author(s): Okizuka Y
  • Journal Title: Genetic Testing and Molecular Biomarkers 13 Pages: 427-431
  • Peer Reviewed
• [Journal Article] Insertion of the IL1RAPL1 gene into the duplication junction of the dystrophin gene (2009)
  • Author(s): Zhang Z
  • Journal Title: Journal of Human Genetics 54 Pages: 466-473
  • Peer Reviewed
• [Journal Article] High incidence of electrocardiogram abnormalities in young patients with duchenne muscular dystrophy. (2008)
  • Author(s): Takami Y, Takeshima Y, Awano H, Okizuka Y, Yagi M, Matsuo M.
  • Journal Title: Pediatr Neurol. 39 Pages: 399-403
• [Journal Article] Dystrophin nonsense mutations can generate alternative rescue transcripts in lymphocytes. (2008)
  • Author(s): Nishiyama A, Takeshima Y, Zhang Z, Habara Y, Tran TH, Yagi M, Matsuo M.
  • Journal Title: Ann Hum Genet. 72 Pages: 717-724
• [Journal Article] Tandem duplications of two separate fragments of the dystrophin gene in a patient with Duchenne muscular dystrophy. (2008)
  • Author(s): Zhang Z, Takeshima Y, Awano H, Nishiyama A, Okizuka Y, Yagi M, Matsuo M.
  • Journal Title: J Hum Genet. 53 Pages: 215-219
  • NAID: 10021248166
• [Journal Article] A strong exonic splicing enhancer in dystrophin exon 19 achieve proper splicing without an upstream polypyrimidine tract. (2008)
  • Author(s): Habara Y, Doshita M, Hirozawa S, Yokono Y, Yagi M, Takeshima Y, Matsuo M.
  • Journal Title: J Biochem. 143 Pages: 303-310
  • NAID: 10024908594
• [Journal Article] High incidence of electrocardiogram abnormalities in young patients with duchenne muscular dystrophy. (2008)
  • Author(s): Takami Y
  • Journal Title: Pediatr Neurol. 39 Pages: 399-403
  • Peer Reviewed
• [Journal Article] Dystrophin nonsense mutations can generate alternative rescue transcripts in lymphocytes. (2008)
  • Author(s): Nishiyama A
  • Journal Title: Ann Hum Genet 72 Pages: 717-724
  • Peer Reviewed
• [Journal Article] Tandem duplications of two separate fragments of the dystrophingene in a patient with Duchenne muscular dystrophy. (2008)
  • Author(s): Zhang Z
  • Journal Title: J Hum Genet. 53 Pages: 215-219
  • Peer Reviewed
• [Presentation] Chemical treatment of muscular dystrophy that enhances skipping of the mutated exon in the dystrophin gene. (2010)
  • Author(s): Nishida A, Kataoka N, Takeshima Y, Yagi M, Awano H, Ota M, Itoh K, Hagiwara M, Matsuo M.
  • Organizer: The American Society of Human Genetics 60th Annual Meeting.
  • Place of Presentation: Washigton, DC.
• [Presentation] Mutation spectrum of the dystrophin gene in 456 Duchienne/Becker muscular dystrophy cases from one Japanese referral center. (2010)
  • Author(s): Takeshima Y, Yagi M, Ota M, Awano H, Yamauchi Y, Malueka RG, Dwianingsih EK, Nishio H, Matsuo M.
  • Organizer: 15th International Congress of The World Muscle Society.
  • Place of Presentation: Kumamoto.
• [Presentation] Antisense RNA/ethylene-bridged nucleic acids chimera induces exon 45 skipping and restores dystrophin expression in myocytes of Duchenne muscular dystrophy. (2010)
  • Author(s): Yagi M, Ota M, Awano H, Takeshima Y, Matsuo M.
  • Organizer: 15th International Congress of The World Muscle Society.
  • Place of Presentation: Kumamoto
• [Presentation] Antisense RNA/ENA chimera against dystrophin exon 45 leads exon 45 skipping followed by dystrophin expression in cells from duchenne muscular dystrophy. (2010)
  • Author(s): Yagi M, Takeshima Y, Awano H, Ota M, Malueka RG, Dwianingsih EK, Nishida A, Lee T, Matsuo M.
  • Organizer: 6th Annual Meeting of the Oligonucleotide Therapeutics Society.
  • Place of Presentation: Southern California
• [Presentation] Long-term administration of antisense oligonucleotide against dystrophin exon 19 for the treatment of Duchenne muscular dystrophy with exon 20 deletion. (2010)
  • Author(s): Matsuo M, Takeshima Y, Yagi M, Awano H.
  • Organizer: XII International Congress on Neuromusular Diseases.
  • Place of Presentation: Naples.
• [Presentation] Antisense RNA/ethylene- bridged nucleic acid chimera induces exon 45 skipping and restores dystrophin expression in DMD muscle cells. (2010)
  • Author(s): Yagi M, Ota M, Awano H, Takeshima Y, Matsuo M.
  • Organizer: XII International Congress on Neuromusular Diseases.
  • Place of Presentation: Naples.
• [Presentation] Genotype-phenotype correlation of the dystrophinopathy cases with small mutations in the dystrophin gene. (2010)
  • Author(s): Awano H, Takeshima Y, Yagi M, Yamauchi Y, Malueka RG, Dwianingsih EK, Matsuo M.
  • Organizer: Pediatric Academic Societies' 2010 Annual Meeting.
  • Place of Presentation: Vancouver.
• [Presentation] 培養筋細胞におけるアンチセンスオリゴヌクレオチドによるエクソン45のスキッピング誘導率は変異によって異なる (2010)
  • Author(s): 八木麻理子
  • Organizer: 第52回日本小児神経学会総会
  • Place of Presentation: 福岡国際会議場(福岡県)
• [Presentation] Duchenne 型筋ジストロフィー分子治療の適応に関する検討 (2009)
  • Author(s): 竹島泰弘
  • Organizer: 第51回日本小児神経学会総会
  • Place of Presentation: 米子コンベンションセンター(鳥取県)
  • Year and Date: 2009-05-29
• [Presentation] Antisense RNA/ethylene-bridged nucleic acids chimera induces exon skipping and restores dystrophin expression in myocytes of Duchenne muscular dystrophy. (2009)
  • Author(s): Yagi M, Yamauchi Y, Awano H, Takeshima Y, Matsuo M.
  • Organizer: The American Society of Human Genetics 59th Annual Meeting.
  • Place of Presentation: Hawaii.
• [Presentation] In vitro splicing analysis reveals that availability of a cryptic splice site is not a determinant for alternative splicing patterns caused by +1G>A mutations in introns of the dystrophin gene. (2009)
  • Author(s): Matsuo M, Habara Y, Takeshima Y, Awano H, Okizuka Y, Zhang Z, Yagi M.
  • Organizer: 14th International Congress of The World Muscle Society.
  • Place of Presentation: Geneva.
• [Presentation] High incidence of outlier from the reading-frame rule in dystrophinopathy patients with duplication mutations in the dystrophin gene. (2009)
  • Author(s): Yagi M, Awano H, Okizuka Y, Takeshima Y, Matsuo M.
  • Organizer: Pediatric Academic Societies' 2009 Annual Meeting.
  • Place of Presentation: Baltimore.
• [Presentation] Mutation spectrum of the dystrophin gene in 433 Japanese dystrophinopathy cases. (2008)
  • Author(s): Takeshima Y, Yagi M, Okizuka Y, Awano H, Zang Z, Saiki K, Matsuo M.
  • Organizer: The American Society of Human Genetics 58th Annual Meeting.
  • Place of Presentation: Philadelphia.
• [Presentation] Splicing errors casued by small mutations in the dystrophin gene are different between lymphocytes and muscle tissues. (2008)
  • Author(s): Okizuka Y, Awano H, Yagi M, Takeshima Y, Matsuo M.
  • Organizer: Pediatric Academic Societies' 2008 Annual Meeting.
  • Place of Presentation: Hawaii.
• [Presentation] A novel retrotransposon that was recently inserted into exon 67 of the dystrophin gene. (2008)
  • Author(s): Awano H, Yagi M, Okizuka Y, Zhang Z, Takeshima Y, Matsuo M.
  • Organizer: Joint 7th Human Genome Organization (HUGO)-Pacific Meeting and the 8th Asia-Pacific Conference on Human Genetics.
  • Place of Presentation: Cebu.
• [Presentation] Mutation spectrum of the dystrophin gene in 433 Japanese dystrophinopathy cases. (2008)
  • Author(s): Takeshima Y
  • Organizer: The American Society of Human Genetics 58th Annual Meeting
  • Place of Presentation: Philadelphia, USA