| Project/Area Number |
18591151
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Kobe University |
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Principal Investigator |
NISHIO Hisahide Kobe University, Graduate School of Medicine, Professor (80189258)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHIMA Yasuhiro Kobe University, Graduate School of Medicine, Assistant professor (40281141)
LEE Myeong, Jin Kobe University, Graduate School of Medicine, Assistant professor (20273766)
綾木 仁 神戸大学, 大学院医学系研究科, 助教授 (80222701)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | spinal muscular atrophy / the SMN1 gene / the SMN2 gene / thetic exon-specific splicing activator / valproic acid / ヒストン脱アセチル化酵素阻害剤 / 転写効率 / スプライシング効率 / エクソン7 / スプライシング / ペプチド核酸 / 遺伝子変異 / Tudorドメイン |
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| Research Abstract |
1. (Background) More than 90% of patients with spinal muscular atrophy (SMA) are homozygous for SMN1 deletion. However, SMN2 is retained in such patients. SMN2 is an almost identical gene to SMN1 and codes the same protein as SMN1 does. The main product of SMN2 is a transcript lacking exon7, producing truncated SMN protein. That is the reason why SMN2 cannot compensate the loss of SMNI in most SMA patients. A single nucleotide change in exon 7, 6C in SMN1 and 6T in SMN2, makes the splicing difference between them. Correction of the exon 7 splicing of SMN2 is now considered as a treatment strategy for SMA.
2. (Synthetic exon-specific splicing activator) Based on the report of Cartegni, et. al., we synthesized a peptide nucleic acid, SMN-PNA (ESSENCE), which may activate splicing of SMN2 exon 7. SMN-PNA had SMN2 exon 7 binding domain and serine-arginine repeat domain. Our study using fibroblast cell line from an SMA patient did not show at all that SMN-PNA corrected the splicing pattern, while in-vitro splicing assay with SMN2EX6-7 plasmid showed some effects on the splicing pattern.
3. (Splicing modulating drug) Valproic acid (VPA) is widely used as an antiepileptic drug. Recently, it has been reported that VPA may increase SMN2 expression and alter its splicing pattern. We tested whether VPA can increase SMN2 gene expression in the fibroblasts from our SMA patients. The effect of VPA (concentration of 0.5-1000 μM) on total transcription level and exon 7-splicing pattern of SMN2 in the fibroblasts was evaluated by quantitative PCR method. VPA did not alter significantly the total transcription level and splicing pattern of SMN2 in this study, suggesting that there are non-responders to VPA treatment.
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