Investigation of fibrotic factors during the exon-skipping therapy using antisense oligonucleotide for muscular dystrophy

• Project/Area Number: 23591495
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kobe University
• Principal Investigator: TAKESHIMA Yasuhiro 神戸大学, 医学(系)研究科(研究院), 教授 (40281141)
• Co-Investigator(Kenkyū-buntansha): YAGI Mariko 神戸大学, 大学院医学研究科, 研究員 (60362787)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: アンチセンスオリゴヌクレオチド / 筋ジストロフィー / エクソンスキッピング / 線維化因子 / アンチセンスオリゴ

Research Abstract

To enhance the effectiveness of antisense oligonucleotide (AO)-mediated exon skipping therapy for Duchenne muscular dystrophy (DMD), the relationship of fibrosis and inflammation to DMD was examined. At the beginning pharmacodynamics of the AO inducing the skipping of exon 45(AO85) was examined using cell-free splicing system, and the EC50 of AO85 was revealed to be 58.0 nM. Then, urinary tetranor PGDM which is major urinary metabolite of prostaglandin (PG) D2 were shown to be increased in DMD patients and became higher with advancing age. It was indicated that PGD2-mediated inflammation plays a role in the pathology of DMD. Furthermore, administration of AO85 to DMD cases resulted in the improvement of some inflammatory cytokines. These results indicated that fibrosis and inflammation were involved in the pathogenesis of DMD, and the modification of these factors was considered as one possible strategy for the enhancement of the effectiveness of AO-mediated exon skipping therapy.

Research Products

• [Journal Article] Differences in carrier frequency between mothers of Duchenne and Becker muscular dystrophy patients (2014)
  • Author(s): Lee T, Takeshima Y, Kusunoki N, Awano H, Yagi M, Matsuo M, and Iijima K
  • Journal Title: J Hum Genet Volume: 59 Pages: 46-50
  • NAID: 40019946811
  • Peer Reviewed
• [Journal Article] Differences in carrier frequency between mothers of Duchenne and Becker muscular dystrophy patients. (2014)
  • Author(s): Lee T, Takeshima Y, Kusunoki N, Awano H, Yagi M, Matsuo M, Iijima K.
  • Journal Title: J Hum Genet. Volume: 59 Pages: 46-50
  • NAID: 40019946811
  • Peer Reviewed
• [Journal Article] Molecular characterization of an X(p21.2;q28) chromosomal inversion in a Duchenne muscular dystrophy patient with mental retardation reveals a novel long non-coding gene on Xq28 (2013)
  • Author(s): Tran TH, Zhang Z, Yagi M, Lee T, Awano H, Nishida A, Okinaga T, Takeshima Y, and Matsuo M
  • Journal Title: J Hum Genet Volume: 58 Pages: 33-9
  • NAID: 10031145898
  • Peer Reviewed
• [Journal Article] Utility of transmural myocardial strain profile for prediction of early left ventricular dysfunction in patients with Duchenne muscular dystrophy (2013)
  • Author(s): Yamamoto T, Tanaka H, Matsumoto K, Lee T, Awano H, Yagi M, Imanishi T, Hayashi N, Takeshima Y, Kawai H, Kawano S, and Hirata K
  • Journal Title: Am J Cardiol Volume: 111 Pages: 902-7
  • Peer Reviewed
• [Journal Article] A prostaglandin D2 metabolite is elevated in the urine of Duchenne muscular dystrophy patients and increases further from 8 years old (2013)
  • Author(s): Nakagawa T, Takeuchi A, Kakiuchi R, Lee T, Yagi M, Awano H, Iijima K, Takeshima Y, Urade Y, and Matsuo M
  • Journal Title: Clin Chim Acta Volume: 423 Pages: 10-4
  • Peer Reviewed
• [Journal Article] A prostaglandin D2 metabolite is elevated in the urine of Duchenne muscular dystrophy patients and increases further from 8 years old. (2013)
  • Author(s): Nakagawa T, Takeuchi A, Kakiuchi R, Lee T, Yagi M, Awano H, Iijima K, Takeshima Y, Urade Y, Matsuo M.
  • Journal Title: Clin Chim Acta. Volume: 423 Pages: 10-4
  • Peer Reviewed
• [Journal Article] Utility of transmural myocardial strain profile for prediction of early left ventricular dysfunction in patients with Duchenne muscular dystrophy. (2013)
  • Author(s): Yamamoto T, Tanaka H, Matsumoto K, Lee T, Awano H, Yagi M, Imanishi T, Hayashi N, Takeshima Y, Kawai H, Kawano S, Hirata K.
  • Journal Title: Am J Cardiol. Volume: 111 Pages: 902-7
  • Peer Reviewed
• [Journal Article] Molecular characterization of an X(p21.2;q28) chromosomal inversion in a Duchenne muscular dystrophy patient with mental retardation reveals a novel long non-coding gene on Xq28. (2013)
  • Author(s): Tran TH, Zhang Z, Yagi M, Lee T, Awano H, Nishida A, Okinaga T, Takeshima Y, Matsuo M.
  • Journal Title: J Hum Genet. Volume: 58 Pages: 33-9
  • NAID: 10031145898
  • Peer Reviewed
• [Journal Article] A prostaglandin D2 metabolite is elevated in the urine of Duchenne muscular dystrophy patients and increases further from 8 years old (2013)
  • Author(s): Taku Nakagawa, Atsuko Takeuchi, Ryohei Kakiuchi, Tomoko Lee, Mariko Yagi, Hiroyuki Awano, Kazumoto Iijima, Yasuhiro Takeshima, Yoshihiro Urade , Masafumi Matsuo
  • Journal Title: Clinica Chimica Acta Volume: in press
  • Peer Reviewed
• [Journal Article] A G-to-T transversion at the splice acceptor site of dystrophin exon 14 shows multiple splicing outcomes that are not exemplified by transition mutations (2012)
  • Author(s): Ota M, Takeshima Y, Nishida A, Awano H, Lee T, Yagi M, and Matsuo M
  • Journal Title: Genet Test Mol Biomarkers Volume: 16 Pages: 3-8
  • Peer Reviewed
• [Journal Article] Optimizing RNA/ENA chimeric antisense oligonucleotides using in vitro splicing (2012)
  • Author(s): Takeshima Y, Yagi M, and Matsuo M
  • Journal Title: Methods Mol Biol Volume: 867 Pages: 131-41
  • Peer Reviewed
• [Journal Article] Categorization of 77 dystrophin exons into 5 groups by a decision tree using indexes of splicing regulatory factors as decision markers (2012)
  • Author(s): Malueka RG, Takaoka Y, Yagi M, Awano H, Lee T, Dwianingsih EK, Nishida A, Takeshima Y, and Matsuo M
  • Journal Title: BMC Genet Volume: 13 Pages: 23-23
  • Peer Reviewed
• [Journal Article] Pathogenic orphan transduction created by a nonreference LINE-1 retrotransposon. (2012)
  • Author(s): Solyom S, Ewing AD, Hancks DC, Takeshima Y, Awano H, Matsuo M, Kazazian H H, Jr.
  • Journal Title: Hum Mutat. Volume: 33(2) Issue: 2 Pages: 369-71
  • DOI: 10.1002/humu.21663
  • Peer Reviewed
• [Journal Article] Categorization of 77 dystrophin exons into 5 groups by a decision tree using indexes of splicing regulatory factors as decision markers (2012)
  • Author(s): Rusdy Ghazali Malueka, Yutaka Takaoka, Mariko Yagi, Hiroyuki Awano, Tomoko Lee, Ery Kus Dwianingsih, Atsushi Nishida, Yasuhiro Takeshima, Masafumi Matsuo
  • Journal Title: BMC Genetics Volume: 13(23) Issue: 1 Pages: 23-23
  • DOI: 10.1186/1471-2156-13-23
  • Peer Reviewed
• [Journal Article] A G-to-T transversion at the splice acceptor site of dystrophin exon 14 shows multiple splicing outcomes that are not exemplified by transition mutations. (2012)
  • Author(s): Ota M, Takeshima Y, Nishida A, Awano H, Lee T, Yagi M, Matsuo M.
  • Journal Title: Genet Test Mol Biomarkers. Volume: 16 Issue: 1 Pages: 3-8
  • DOI: 10.1089/gtmb.2010.0276
  • Peer Reviewed
• [Journal Article] Categorization of 77 dystrophin exons into 5 groups by a decision tree using indexes of splicing regulatory factors as decision markers. (2012)
  • Author(s): Malueka RG, Takaoka Y, Yagi M, Awano H, Lee T, Dwianingsih EK, Nishida A, Takeshima Y, Matsuo M.
  • Journal Title: BMC Genet. Volume: 13 Pages: 23-23
  • Peer Reviewed
• [Journal Article] Optimizing RNA/ENA Chimeric Antisense Oligonucleotides Using In Vitro Splicing. (2012)
  • Author(s): Takeshima Y, Yagi M, Matsuo M.
  • Journal Title: Methods Mol Biol. Volume: 867 Pages: 131-41
• [Journal Article] Antisense oligonucleotide induced dystrophin exon 45 skipping at a low half-maximal effective concentration in a cell-free splicing system (2011)
  • Author(s): Malueka RG, Yagi M, Awano H, Lee T, Dwianingsih EK, Nishida A, Takeshima Y, and Matsuo M
  • Journal Title: Nucleic Acid Ther Volume: 21 Pages: 347-53
  • Peer Reviewed
• [Presentation] The carrier frequency in the mothers of 158 Japanese cases with Duchenne/Becker muscular dystrophy (2013)
  • Author(s): Lee T, Kusunoki N, Yagi M, Takeshima Y, Matsuo M. and Iijima K
  • Organizer: European Human Genetics Conference 2013
  • Place of Presentation: Paris, France
• [Presentation] Three-dimensional gait analysis of Duchenne muscular dystrophy; a trial to evaluate the therapeutic effect of RNA/ENA chimera antisense oligonucleotide that induces dystrophin exon 45 skipping (2013)
  • Author(s): Takeshima Y, Yagi M, Lee T, Kusunoki N, Ojima I, Minami S, Asai T, Nakagawa A, Iijima K, and Matsuo M
  • Organizer: 18th International Congress of the World Muscle Society
  • Place of Presentation: California, USA
• [Presentation] A prostaglandin D2 metabolite is elevated in the urine samples of patients with Duchenne muscular dystrophy (2013)
  • Author(s): Nakagawa T, Takeuchi A, Kakiuchi R, Lee T, Yagi M, Awano H, Iijima K, Takeshima Y, Urade Y, and Matsuo M
  • Organizer: 18th International Congress of the World Muscle Society
  • Place of Presentation: California, USA
• [Presentation] A novel splicing silencer generated by dystrophin exon 45 deletion could explain exon 44 skipping that modifies dystrophinopathy (2013)
  • Author(s): Matsuo M, Dwianingsih EM, Malueka RG, Nishida A, Lee T, Yagi M, Iijima K, and Takeshima Y
  • Organizer: American Society of Human Genetics, the 63^<rd> Annual Meeting
  • Place of Presentation: Boston, USA
• [Presentation] The carrier frequency in the mothers of 158 Japanese cases with Duchenne/Becker muscular dystrophy. (2013)
  • Author(s): Lee T, Kusunoki N, Yagi M, Takeshima Y, Matsuo M, Iijima K.
  • Organizer: European Human Genetics Conference 2013
  • Place of Presentation: Paris
• [Presentation] Three-dimensional gait analysis of Duchenne muscular dystrophy; a trial to evaluate the therapeutic effect of RNA/ENA chimera antisense oligonucleotide that induces dystrophin exon 45 skipping. (2013)
  • Author(s): Takeshima Y, Yagi M, Lee T, Kusunoki N, Ojima I, Minami S, Asai T, Nakagawa A, Iijima K, Matsuo M.
  • Organizer: 18th International Congress of the World Muscle Society
  • Place of Presentation: California
• [Presentation] A prostaglandin D2 metabolite is elevated in the urine samples of patients with Duchenne muscular dystrophy. (2013)
  • Author(s): Nakagawa T, Takeuchi A, Kakiuchi R, Lee T, Yagi M, Awano H, Iijima K, Takeshima Y, Urade Y, Matsuo M.
  • Organizer: 18th International Congress of the World Muscle Society
  • Place of Presentation: California
• [Presentation] A novel splicing silencer generated by dystrophin exon 45 deletion could explain exon 44 skipping that modifies dystrophinopathy. (2013)
  • Author(s): Matsuo M, Dwianingsih EM, Maliueka RG, Nishida A, Lee T, Yagi M, Iijima K, Takeshima Y.
  • Organizer: American Society of Human Genetics
  • Place of Presentation: Boston
• [Presentation] A small chemical, TG003, enhances skipping of mutated dystrophin exons : the third example revealing a decrease of exonic splicing enhancer density in common (2012)
  • Author(s): Nishida A, Takeshima Y, Kataoka N, Yagi M, Awano H, Lee T, Iijima K, Hagiwara M, and Matsuo M
  • Organizer: The American Society of Human Genetics, the 62^<nd> Annual Meeting
  • Place of Presentation: San Francisco, USA
• [Presentation] Dystrophin Gene Mutations in Three Dystrophynopathy Patients with Severe Cardiomyopathy. (2012)
  • Author(s): Awano H, Lee T, Yagi M, Takeshima Y, Matsuo M, Iijima K.
  • Organizer: Pediatric Academic Societies Annual Meeting
  • Place of Presentation: Boston
• [Presentation] A small chemical, TG003, enhances skipping of mutated dystrophin exons: the third example revealing a decrease of exonic splicing enhancer density in common. (2012)
  • Author(s): Nishida A, Takeshima Y, Kataoka N, Yagi M, Awano H, Lee T, Iijima K, Hagiwara M, Matsuo M.
  • Organizer: The American Society of Human Genetics, the 62th Annual Meeting
  • Place of Presentation: San Francisco
• [Presentation] Antisense RNA/ ethylene-bridged nucleic acid chimera induces exon skipping in cultured myocytes from DMD patients with 6 different deletion mutations (2011)
  • Author(s): Yagi M, Lee T, Awano H, Takeshima Y, and Matsuo M
  • Organizer: The American Society of Human Genetics 61st Annual Meeting
  • Place of Presentation: Montral, Canada
• [Presentation] Antisense RNA/Ethylene-bridged nucleic acid chimera induces exon 45 skipping in cultured myocytes from DMD patients with 6 different deletion mutations (2011)
  • Author(s): Yagi M, Lee T, Awano H, Takeshima Y, Matsuo M
  • Organizer: The American Society of Human Genetics 61st Annual Meeting
  • Place of Presentation: Montreal