Drug treatment for spinal muscular atrophy activating signal transduction pathways

• Project/Area Number: 25461549
• 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: Nishio Hisahide 神戸大学, 医学(系)研究科(研究院), 教授 (80189258)
• Co-Investigator(Kenkyū-buntansha): NISHIMURA Noriyuki 神戸大学, 大学院医学研究科, 教授 (00322719) ; MORIKAWA Satoru 神戸大学, 大学院医学研究科, 講師 (50457074)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 脊髄性筋萎縮症 / ＳＭＮ１遺伝子 / ＳＭＮ２遺伝子 / ＳＭＮ蛋白 / サルブタモール / 交感神経作動薬 / ユビキチン化 / SMN1遺伝子 / SMN2遺伝子 / SMN蛋白 / 交感神経β2受容体 / 交感神経β2受容体作動薬

Outline of Final Research Achievements

Spinal muscular atrophy (SMA) is a genetic neuromuscular disorder. SMA is caused by the loss of the SMN1 gene. Although the SMN2 gene also produces SMN protein, it is not enough to compensate for the loss of the SMN1 gene. Salbutamol (adrenergic drug) was reported to improve SMA symptoms. To clarify the mechanism, we treated SMA fibroblasts lacking the SMN1 gene with salbutamol, and analyzed SMN2 mRNA and SMN protein levels. Salbutamol increased SMN protein levels in a dose-dependent manner, although SMN2 mRNA levels were not changed. The salbutamol-induced increase in SMN protein level was blocked by PKA inhibitor and deubiquitinase inhibitor. Immunoprecipitation assay using HeLa cells showed that salbutamol decreased ubiquitinated SMN protein levels, suggesting that salbutamol inhibited ubiquitination. These findings suggest that salbutamol increases SMN protein levels in SMA cells by inhibiting ubiquitin-mediated SMN degradation via activating β2-adrenergic receptor-PKA pathways.

Research Products

• [Journal Article] A new method for SMN1 and hybrid SMN gene analysis in spinal muscular atrophy using long-range PCR followed by sequencing. (2015)
  • Author(s): Kubo Y, Nishio H, Saito K
  • Journal Title: J Hum Genet Volume: 60 Issue: 5 Pages: 233-239
  • DOI: 10.1038/jhg.2015.16
  • NAID: 40020462572
  • Peer Reviewed / Open Access
• [Journal Article] Two Japanese patients with SMA type 1 suggest that axonal-SMN may not modify the disease severity. (2015)
  • Author(s): Yamada H, Nishida Y, Maihara T, Sa'adah N, Harahap NI, Nurputra DK, Ar Rochmah M, Nishimura N, Saito T, Kubo Y, Saito K, Nishio H.
  • Journal Title: Pediatr Neurol. Volume: 52 Issue: 6 Pages: 638-641
  • DOI: 10.1016/j.pediatrneurol.2015.02.023
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Rapid, Accurate and Simple Screening Method for Spinal Muscular Atrophy: High-Resolution Melting Analysis Using Dried Blood Spots on Filter Paper. (2015)
  • Author(s): Sa'adah N, Harahap NI, Nurputra DK, Rochmah MA, Morikawa S, Nishimura N, Sadewa AH, Astuti I, Haryana SM, Saito T, Saito K, Nishio H.
  • Journal Title: Clin Lab. Volume: 61 Issue: 05+06/2015 Pages: 575-580
  • DOI: 10.7754/clin.lab.2014.141008
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Trinucleotide insertion in the SMN2 promoter may not be related to the clinical phenotype of SMA. (2015)
  • Author(s): Harahap NI, Takeuchi A, Yusoff S, Tominaga K, Okinaga T, Kitai Y, Takarada T, Kubo Y, Saito K, Sa'adah N, Nurputra DK, Nishimura N, Saito T, Nishio H.
  • Journal Title: Brain Dev. Volume: 37 Issue: 7 Pages: 669-676
  • DOI: 10.1016/j.braindev.2014.10.006
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Salbutamol inhibits ubiquitin-mediated survival motor neuron protein degradation in spinal muscular atrophy cells. (2015)
  • Author(s): Harahap NI, Nurputra DK, Rochmah MA, Shima A, Morisada N, Takarada T, Takeuchi A, Tohyama Y Yanagisawa S, Nishio H
  • Journal Title: Biochemistry and Biophysics Reports Volume: 4 Pages: 351-356
  • DOI: 10.1016/j.bbrep.2015.10.012
  • Peer Reviewed / Open Access
• [Journal Article] SMA Screening System Using Dried Blood Spots on Filter Paper: Application of COP-PCR to the SMN1 Deletion Test. (2015)
  • Author(s): Kato N, Sa'Adah N, Ar Rochmah M, Harahap NI, Nurputra DK, Sato H, Sadewa AH, Astuti I, Haryana SM, Saito T, Saito K, Nishimura N, Nishio H, Takeuchi A.
  • Journal Title: Kobe J Med Sci. Volume: 60
  • NAID: 110009871896
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Study of valproic acid for patients with spinal muscular atrophy. (2015)
  • Author(s): Saito T, Nurputra DK, Harahap NI, Harahap IS, Yamamoto H, Muneshige E, Nisizono H, Matsumura T, Fujimura H, Sakoda S, Saito K, Nishio H.
  • Journal Title: Neurology and Clinical Neuroscience Volume: 3 Issue: 2 Pages: 49-57
  • DOI: 10.1111/ncn3.140
  • Peer Reviewed
• [Journal Article] Intragenic mutations in SMN1may contribute more significantly to clinical severity than SMN2 copy numbers in some spinal muscular atrophy (SMA) patients (2014)
  • Author(s): Yamamoto T, Sato H, Lai PS, Nurputra DK, Harahap NIF, Morikawa S, Nishimura N, Kurashige T, Tomohiko O, Nakajima H, Yamada H, Nishida Y, Toda S, Takanashi J, Takeuchi A, Tohyama Y, Kubo Y, Saito K, Takeshima Y, Matsuo M, Nishio H
  • Journal Title: Brain Dev Volume: 未 Issue: 10 Pages: 914-920
  • DOI: 10.1016/j.braindev.2013.11.009
  • Peer Reviewed
• [Journal Article] 脊髄性筋萎縮症 遺伝子診断から治療戦略まで (2014)
  • Author(s): 西尾 久英
  • Journal Title: 日本小児科学会雑誌 Volume: 118 Pages: 1315-1323
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] PLS3 expression and SMA phenotype: a commentary on correlation of PLS3 expression with disease severity in children with spinal muscular atrophy. (2014)
  • Author(s): Nishio H.
  • Journal Title: J Hum Genet. Volume: 59 Issue: 2 Pages: 64-65
  • DOI: 10.1038/jhg.2013.124
  • Peer Reviewed
• [Journal Article] Molecular pathology of Sandhoff disease with p.Arg505Gln in HEXB: application of simulation analysis. (2013)
  • Author(s): Yasui N, Takaoka Y, Nishio H, Nurputra DK, Sekiguchi K, Hamaguchi H, Kowa H, Maeda E, Sugano A, Miura K, Sakaeda T, Kanda F, Toda T.
  • Journal Title: J Hum Genet Volume: 58 Issue: 9 Pages: 611-617
  • DOI: 10.1038/jhg.2013.68
  • NAID: 10031195298
  • Peer Reviewed
• [Journal Article] Spinal Muscular Atrophy: From Gene Discovery to Clinical Trials (2013)
  • Author(s): Nurputra DK, Lai PS, Harahap NI, Morikawa S, Yamamoto T, Nishimura N, Kubo Y, Takeuchi A, Saito T, Takeshima Y, Tohyama Y, Tay SK, Low PS, Saito K, Nishio H
  • Journal Title: Ann Hum Genet. Volume: 77 Issue: 5 Pages: 435-463
  • DOI: 10.1111/ahg.12031
  • Peer Reviewed
• [Journal Article] 劇症肝不全を発症した脊髄性筋萎縮症の1例 (2013)
  • Author(s): 羽深 理恵, 鈴木 俊明, 長谷川 博也, 唐澤 環, 金子 詩子, 池住 洋平, 大橋 伯, 赤坂 紀幸, 遠山 潤, 西尾 久英, 齋藤 昭彦
  • Journal Title: 日本小児科学会雑誌 Volume: 117 Pages: 1031-1036
  • NAID: 10031184018
  • Peer Reviewed
• [Presentation] Gender effects on the severity of spinal muscular atrophy (SMA) in 286 Japanese patients diagnosed in 1996-2015. (2016)
  • Author(s): Shima A, Ar Rochmah M, Morisada N, Yanagisawa Y, Harahap NI, Saito T, Umeno Aiko, Kaneko K, Saito K, Iijima K, Nishio H
  • Organizer: 第13回国際人類遺伝学会
  • Place of Presentation: 京都, 国立京都国際会館
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] Sulbutamol inhibits ubiquitin-mediated survival motor neuron protein degradation in spinal muscular atrophy cells. (2016)
  • Author(s): Harahap NI, Nurputra DK, Ar Rochmah M, Shima A, Morisada N, Takarada T, Takeuchi A, Tohyama Y, Yanagisawa Y, Nishio H.
  • Organizer: 第13回国際人類遺伝学会
  • Place of Presentation: 京都, 国立京都国際会館
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] 脊髄性筋萎縮症の重症度を規定する因子についての検討 (2015)
  • Author(s): 森貞直哉, 島亜衣, Mawaddah Ar Rochmah, 叶明娟, 飯島一誠, 西尾久英
  • Organizer: 日本人類遺伝学会第60回大会
  • Place of Presentation: 東京, 京王プラザホテル
  • Year and Date: 2015-10-14
• [Presentation] 小児期発症脊髄性筋萎縮症に対するバルプロ酸ナトリウム多施設共同医師主導治験 (2015)
  • Author(s): 斎藤加代子, 荒川玲子, 齊藤利雄, 西尾久英
  • Organizer: 第57回日本小児神経学会学術集会
  • Place of Presentation: 大阪, 帝国ホテル
  • Year and Date: 2015-05-28
• [Presentation] HDAC6, a new candidate for therapeutic target of Spinal Muscular Atrophy (SMA) (2015)
  • Author(s): Dian K Nurputra, Morita H, Harahap NI, Rochmah MA, Tohyama Y, Nishio H.
  • Organizer: The 11th Asian Society for Pediatric Research
  • Place of Presentation: 大阪, グランキューブ大阪
  • Year and Date: 2015-04-15
  • Int'l Joint Research
• [Presentation] 脊髄性筋萎縮症におけるSMN1、SMN2、NAIP、GTF2H2、SERF1遺伝子解析 (2014)
  • Author(s): 野口依子, 中町祐司, 林伸英, 河野誠司, 中川卓, 西尾久英
  • Organizer: 第61回日本臨床検査医学会学術集会
  • Place of Presentation: 福岡市, 福岡国際会議場
  • Year and Date: 2014-11-22 – 2014-11-25
• [Presentation] HDAC6, a new therapeutic target of SMA, work in the downstream pathway of cytoskeleton dynamics. (2014)
  • Author(s): Nurputra DK， Morita H，Tohyama Y, Nishio H.
  • Organizer: 日本人類遺伝学会第59回大会
  • Place of Presentation: 東京都, タワーホール船堀
  • Year and Date: 2014-11-19 – 2014-11-22
• [Presentation] SMN1遺伝子exon6にframeshift変異を認めた脊髄性筋萎縮症1型の一例 (2014)
  • Author(s): 山田博之, 西田吉伸, 松本貴子, 毎原敏郎, 山本友人, 西尾久英
  • Organizer: 第56回小児神経学会総会
  • Place of Presentation: 浜松市, アクトシティ浜松
  • Year and Date: 2014-05-29 – 2014-05-31
• [Presentation] SMN is essential for the HDAC6 mediated tubulin-deacetylation in fibroblasts. (2014)
  • Author(s): Nurputra DK， Morita H， Nishio H， Tohyama Y.
  • Organizer: 第61回日本生化学会近畿支部例会
  • Place of Presentation: 京都市, 京都産業大学
  • Year and Date: 2014-05-17
• [Presentation] Abnormal acetylation status of α-tubulin in fibroblasts derived from SMA patients. (2013)
  • Author(s): Nurputra DK, Morita H, Nishio H, Tohyama Y.
  • Organizer: 第60回日本生化学会近畿支部例会
  • Place of Presentation: 吹田市、大阪大学 吹田キャンパス
• [Presentation] 軽症で経過しているSMN1遺伝子新規点変異(c.5C>T)を認めたspinal muscular atrophy type IIIの16歳男性例 (2013)
  • Author(s): 中森正博, 大下智彦, 倉重毅志, 上野弘貴, 越智一秀, 丸山博文, 西尾久英, 松本昌泰
  • Organizer: 第94回日本神経学会中国・四国地方会
  • Place of Presentation: 松山市、松山市総合コミュニティセンター
• [Presentation] 神経筋疾患とIGF-1: 脊髄性筋萎縮症治療法の確立に向けて。 (2013)
  • Author(s): 西尾久英
  • Organizer: 第47回兵庫内分泌研究会
  • Place of Presentation: 神戸市、神戸ポートピアホテル
  • Invited
• [Presentation] 分裂酵母モデル系を用いた脊髄性筋萎縮症の原因タンパク質Smn1の機能解明 (2013)
  • Author(s): 西島数人, 加藤俊明, 馬艶, 西尾久英
  • Organizer: 第124回日本薬理学会近畿部会
  • Place of Presentation: 京都、京都ガーデンパレス
• [Book] 別冊 日本臨床 新領域別症候群シリーズNo.27 (2014)
  • Author(s): 西尾久英
  • Total Pages: 903
  • Publisher: 日本臨床社