Dystrophin intron retention analysis to identify new targets for Antisense Oligonucleotide mediated RNA modulation in Rhabdomyosarcoma

2018 Fiscal Year Annual Research Report

• Project/Area Number: 16K07216
• Research Institution: Kobe University
• Principal Investigator: ニバ タベ・エマ・エコ 神戸大学, 医学研究科, 助教 (00727810)
• Co-Investigator(Kenkyū-buntansha): 松尾 雅文 神戸学院大学, 総合リハビリテーション学部, 特命教授 (10157266)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: splicing, / intron retention, / dystrophin, / antisense chemistry, / rhabdomyosarcoma, / tumor, / tumor suppressor gene,

Outline of Annual Research Achievements

Dystrophin is regarded as a potential tumor suppressor gene because it was shown to be mutated in a variety of rhabdomyosarcoma. Also DMD patients sometimes develop some kind of tumor. Therefore, this project was intended to understand the significance of dystrophin as a tumor suppressor in rhabdomyosarcoma.
In the previous year, the applicants designed an antisense oligonucleotide to alter the splicing of intron 40 retention in rhabdomyosarcoma. They could confirm the abolishment of the intron 40 retention in rhabdomyosarcoma. Also, they observed that the antisense could reduce the growth of rhabdomyosarcoma after 72 hours of incubation. In addition, one carboxyl terminal isoform of dystrophin also showed increase when rhabdomyosarcoma was treated with the antisense.
They also confirmed that this antisense was specific to the cancer-type. They tested in other cancer-types such as colon cancer that showed also retention of intron 40. However, this antisense could not eliminate the intron 40 retention and had no effect on the growth of the colon cancer.
Species specificity was also tested, however, there was no effect on mouse generated cancer cells.
The applicants are now preparing the data for submission.
Finally, the applicants showed that intron retention could be a factor of tumor formation because, growth was reduced after treatment with intron removal agent, antisense oligonucleotide.

Research Products

• [Journal Article] Detection of Dystrophin Dp71 in Human Skeletal Muscle Using an Automated Capillary Western Assay System (2018)
  • Author(s): Kawaguchi T*, Niba ETE*, Rani AQM, Onishi Y, Koizumi M, Awano
  • Journal Title: International Journal of Molecular Science Volume: 19 (6) Pages: E1546
  • DOI: doi:10.3390/ijms19061546
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Intron-retained transcripts of the spinal muscular atrophy genes, SMN1 and SMN2 (2018)
  • Author(s): Harahap NIF, Niba ETE, Ar Rochmah M, Wijaya YOS, Saito T, Saito K, Awano H, Morioka I, Iijima K, Lai PS, Motsuo M, Nishio H, Shinohara M
  • Journal Title: Brain & Development Volume: 40 (8) Pages: 670-677
  • DOI: doi: 10.1016/j.braindev.2018.03.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] DMD transcription profiling in spontaneously developed tumor from three mdx mice revealed extensive intron retentions and Dp71 isoform expression (2018)
  • Author(s): Emma E.T. Niba, Noriyuki Nishimura, Satoru Takafuji, Khin Kyae Mon Thwin, Nobuyuki Yamamoto, Hiroyuki Awano, Shoji Fukushima, Kyoko Itoh, Hisahide Nishio, Masafumi Matsuo
  • Organizer: American Society of Cell Biology
  • Int'l Joint Research
• [Presentation] Glioma from a DMD patient exhibits differential splicing pattern including exon 71 skipping and intron 40 retention (2018)
  • Author(s): : Emma E.T. Niba, Hiroyuki Awano, Masashi Nagai, Masaaki Taniguchi, Rani Adul Qawee, Masakazu Shinohara, Hisahide Nishio, Masafumi Matsuo
  • Organizer: Molecular Biology Society of Japan
  • Int'l Joint Research
• [Presentation] Spontaneous development of spindle cell sarcoma in mdx mice (2018)
  • Author(s): : Satoru Takafuji, Emma Niba, Khin Kyae Mon Thwin, Nobuyuki Yamamoto, Hiroyuki Awano, Suguru Uemura, Takeshi Mori, Shoji Fukushima, Kyoko Itoh, Hisahide Nishio, Masafumi Matsuo, Kazumoto Iijima, Noriyuki Nishimura
  • Organizer: International Society of Pediatric Oncology
  • Int'l Joint Research