Development of Newborn Screening System for Spinal Muscular Atrophy

• Project/Area Number: 20K08197
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kobe Gakuin University
• Principal Investigator: Nishio Hisahide 神戸学院大学, 総合リハビリテーション学部, 教授 (80189258)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 脊髄性筋萎縮症 / 新生児スクリーニング / 乾燥血液ろ紙検体 / SMN1遺伝子欠失 / SMN1遺伝子内変異 / スクリーニング / 乾燥濾紙血 / 乾燥濾紙唾液 / 偽陽性 / ヘパリン / 乾燥濾紙血液 / 乾燥血液濾紙 / 保因者

Outline of Research at the Start

現在開発中のシステムは、「乾燥濾紙血検体をパンチして得た濾紙をそのままPCRチューブに入れて、通常のPCR法で事前増幅を行う」段階と、「そのPCR産物中にSMN1遺伝子が存在しているかどうかを、SMN1遺伝子を特異的に増幅するリアルタイムmCOP-PCR法を用いて確認する」の2つのプロセスからなる。このSMAスクリーニング・システムを完成させ、実際の臨床に応用し、家系内に患者を持つハイリスク家族を対象とするSMAスクリーニングと、一般人口集団の新生児を対象とするSMAスクリーニングを実施する。

Outline of Final Research Achievements

Spinal muscular atrophy (SMA) is a hereditary motor neuron disease that occurs very frequently in Japan. The purpose of this study is to establish a diagnostic test system for SMA using dried filter paper blood and apply it to newborn screening for SMA. The following results were obtained in this research. (1) We developed a screening method for SMN1 gene deletion using dried blood on filter paper. (2) We developed a method for detecting SMN1 intragenic mutations. (3) We have elucidated the cause of false positives in the SMA newborn screening test and found that they are related to heparin, which is used to treat neonatal disease.

Academic Significance and Societal Importance of the Research Achievements

脊髄性筋萎縮症（spinal muscular atrophy: SMA）は、全身の筋萎縮と筋力低下を主症状とする遺伝性運動ニューロン病である。発生頻度は1～2万出生に1人と高く、保因者頻度は50～7人あたり1人と予測されている。1995年に染色体5q13領域のSMN1遺伝子が疾患遺伝子であることが報告され、その後の研究でSMN1遺伝子の近傍に存在するSMN2遺伝子が重症度を規定する遺伝子であることが明らかになった。最近、有効な治療薬が開発されて、しかも早期治療が有効であることが明らかになった。SMAは、臨床症状だけで診断するのが困難である。早期診断のシステム構築が喫緊の課題となった。

Research Products

• [Journal Article] Spinal muscular atrophy type 2 patient who survived 61 years: an autopsy case report. (2022)
  • Author(s): Yamadera M, Saito T, Shinohara M, Nishio H, Murayama S, Fujimura H.
  • Journal Title: Neuropathology Volume: 42 Issue: 2 Pages: 141-146
  • DOI: 10.1111/neup.12784
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High Concentration or Combined Treatment of Antisense Oligonucleotides for Spinal Muscular Atrophy Perturbed SMN2 Splicing in Patient Fibroblasts (2022)
  • Author(s): Wijaya Yogik Onky Silvana、Niba Emma Tabe Eko、Nishio Hisahide、Okamoto Kentaro、Awano Hiroyuki、Saito Toshio、Takeshima Yasuhiro、Shinohara Masakazu
  • Journal Title: Genes Volume: 13 Issue: 4 Pages: 685-685
  • DOI: 10.3390/genes13040685
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Changes in the Incidence of Infantile Spinal Muscular Atrophy in Shikoku, Japan between 2011 and 2020. (2022)
  • Author(s): Okamoto K, Nishio H, Motoki T, Jogamoto T, Aibara K, Kondo Y, Kawamura K, Konishi Y, Tokorodani C, Nishiuchi R, Eguchi M.
  • Journal Title: Int J Neonatal Screen Volume: 8 Issue: 4 Pages: 52-52
  • DOI: 10.3390/ijns8040052
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PCR-Based Screening of Spinal Muscular Atrophy for Newborn Infants in Hyogo Prefecture, Japan (2022)
  • Author(s): Noguchi Yoriko、Bo Ryosuke、Nishio Hisahide、Matsumoto Hisayuki、Matsui Keiji、Yano Yoshihiko、Sugawara Masami、Ueda Go、Wijaya Yogik Onky Silvana、Niba Emma Tabe Eko、Shinohara Masakazu、Bouike Yoshihiro、Takeuchi Atsuko、Okamoto Kentaro、Saito Toshio、Shimomura Hideki、Lee Tomoko、Takeshima Yasuhiro....Awano Hiroyuki
  • Journal Title: Genes Volume: 13 Issue: 11 Pages: 2110-2110
  • DOI: 10.3390/genes13112110
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Stability and Oligomerization of Mutated SMN Protein Determine Clinical Severity of Spinal Muscular Atrophy (2021)
  • Author(s): Niba ETE, Nishio H, Wijaya YOS, Ar Rochmah M, Takarada T, Takeuchi A, Kimizu T, Okamoto K, Saito T, Awano H, Takeshima Y, Shinohara M.
  • Journal Title: Genes (Basel) Volume: 13 Issue: 2 Pages: 205-205
  • DOI: 10.3390/genes13020205
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Dried Blood Spot Screening System for Spinal Muscular Atrophy with Allele-Specific Polymerase Chain Reaction and Melting Peak Analysis (2021)
  • Author(s): Wijaya Yogik Onky Silvana、Nishio Hisahide、Niba Emma Tabe Eko、Shiroshita Tomoyoshi、Kato Masako、Bouike Yoshihiro、Tode Chisato、Ar Rochmah Mawaddah、Harahap Nur Imma Fatimah、Nurputra Dian Kesumapramudya、Okamoto Kentaro、Saito Toshio、Takeuchi Atsuko、Lai Poh San、Yamaguchi Seiji、Shinohara Masakazu
  • Journal Title: Genetic Testing and Molecular Biomarkers Volume: 25 Issue: 4 Pages: 293-301
  • DOI: 10.1089/gtmb.2020.0312
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Detection of Spinal Muscular Atrophy Patients Using Dried Saliva Spots. (2021)
  • Author(s): Wijaya YOS, Nishio H, Niba ETE, Okamoto K, Shintaku H, Takeshima Y, Saito T, Shinohara M, Awano H.
  • Journal Title: Genes (Basel) Volume: 12 Issue: 10 Pages: 1621-1621
  • DOI: 10.3390/genes12101621
  • NAID: 120007169863
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Spinal Muscular Atrophy: Diagnosis, Incidence, and Newborn Screening in Japan. (2021)
  • Author(s): Kimizu T, Ida S, Okamoto K, Awano H, Niba ETE, Wijaya YOS, Okazaki S, Shimomura H, Lee T, Tominaga K, Nabatame S, Saito T, Hamazaki T, Sakai N, Saito K, Shintaku H, Nozu K, Takeshima Y, Iijima K, Nishio H, Shinohara M.
  • Journal Title: Int J Neonatal Screen. Volume: 7 Issue: 3 Pages: 45-45
  • DOI: 10.3390/ijns7030045
  • NAID: 120007159914
• [Journal Article] Phenotypes of SMA patients retaining SMN1 with intragenic mutation (2021)
  • Author(s): Wijaya Yogik Onky Silvana、Nishio Hisahide、Shinohara Masakazu et al.
  • Journal Title: Brain and Development Volume: － Issue: 7 Pages: 745-758
  • DOI: 10.1016/j.braindev.2021.03.006
  • Peer Reviewed
• [Journal Article] A case of spinal muscular atrophy type 0 treated with nusinersen from 10 weeks after birth (2021)
  • Author(s): 木水 友一、西尾 久英、鈴木 保宏、五嶋 嶺、水谷 聡志、中島 健、池田 妙、最上 友紀子、柳原 恵子、沖永 剛志
  • Journal Title: NO TO HATTATSU Volume: 53 Issue: 2 Pages: 142-145
  • DOI: 10.11251/ojjscn.53.142
  • NAID: 130008028463
  • ISSN: 0029-0831, 1884-7668
  • Peer Reviewed
• [Journal Article] Assessment of Spinal Muscular Atrophy Carrier Status by Determining SMN1 Copy Number Using Dried Blood Spots (2020)
  • Author(s): Wijaya YOS, Purevsuren J, Harahap NIF, Niba ETE, Bouike Y, Nurputra DK, Rochmah MA, Thursina C, Hapsara S, Yamaguchi S, Nishio H, Shinohara M.
  • Journal Title: Int J Neonatal Screen. Volume: 29 Issue: 2 Pages: 43-55
  • DOI: 10.3390/ijns6020043
  • Peer Reviewed / Open Access / Int'l Joint Research