Elucidation of a mechanism of tissue-specific alternative splicing by using analysis of splicing order.

• Project/Area Number: 26860793
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Pediatrics
• Research Institution: Gifu University
• Principal Investigator: HORI Tomohiro 岐阜大学, 医学部附属病院, 助教 (90456525)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: スプライシングオーダー / 選択的スプライシング / 遺伝学 / スプライシング

Outline of Final Research Achievements

Our strategy was to use RT-PCR of hnRNA to detect the presence or absence of spliced exon clusters in RNA intermediates (SECRIs) comprising sequential exons. The splice donor site mutation c.1248+5g>a (IVS13) of the OXCT1 gene resulted predominantly in skipping of exons 12 and 13 in skin fibroblasts from a patient with succinyl-CoA:3-ketoacid CoA transferase (SCOT) deficiency. We compared hnRNA intermediates between controls' and the patient's fibroblasts derived from skin, kidney and liver. In controls, intron 11 was the last intron to be spliced and the removal of intron 12 was also rather slow and occurred after the removal of intron 13 in a major pathway. The mutation in the patient’s cells resulted in retention of intron 13, thus causing the retention of introns 12 and 11. Now, we have been conducting studies about splicing order in fibroblasts derived from hepatocyte and the gene responsible for mitochondrial acetoacetyl-CoA thiolase deficiency.

Research Products

• [Journal Article] Inborn errors of ketone body utilization (2015)
  • Author(s): Hori T, Yamaguchi S, Shintaku H, Horikawa R, Shigematsu Y, Hakayanagi M, Fukao T
  • Journal Title: Pediatr Int Volume: 57 Issue: 1 Pages: 51-48
  • DOI: 10.1111/ped.12585
  • Peer Reviewed
• [Journal Article] The first case in Asia of 2-methyl-3-hydroxybutyryl-CoA dehydrogenase deficiency (HSD10 disease) with atypical presentation. (2014)
  • Author(s): Fukao T, Akiba K, Goto M, Kuwayama N, Morita M, Hori T, Aoyama Y, Venkatesan R, Wierenga R, Moriyama Y, Hashimoto T, Usuda N, Murayama K, Ohtake A, Hasegawa Y, ShigematsuY, Hasegawa Y.
  • Journal Title: J Hum Genet Volume: 59 Issue: 11 Pages: 609-614
  • DOI: 10.1038/jhg.2014.79
  • NAID: 40020269650
  • Peer Reviewed
• [Journal Article] Ketone body metabolism and its defects. (2014)
  • Author(s): Fukao T, Mitchell G, Saas JO, Hori T, Orii K, Aoyama Y
  • Journal Title: J Inherited Metab Dis Volume: 37 Issue: 4 Pages: 541-551
  • DOI: 10.1007/s10545-014-9704-9
  • Peer Reviewed / Open Access
• [Presentation] バセドウ病の治療経過中に発症した横紋筋融解症の女児例 (2014)
  • Author(s): 堀友博，松本英樹，大塚博樹，桑原秀次，川本典生，深尾敏幸
  • Organizer: 第48回日本小児内分泌学会学術集会
  • Place of Presentation: 浜松
  • Year and Date: 2014-09-25 – 2014-09-27