DNA methylation-based forensic age prediction in Japanese populations using locked nucleic acid.

• Project/Area Number: 19K19477
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 58040:Forensics medicine-related
• Research Institution: Asahikawa Medical College
• Principal Investigator: Isozaki Shotaro 旭川医科大学, 大学病院, 医員 (60834504)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: Locked Nucleic Acid / メチル化 / 人工核酸 / LNA / 年齢推定 / 法医学 / 法医遺伝学 / DNA

Outline of Research at the Start

法医鑑定における年齢推定は、歯牙や骨の形態学的な特徴により行われるが、個体差や測定者間誤差が存在することから、異なる年齢推定法の確立が望まれている。近年、DNAのメチル化率に着目した新たな年齢推定法の研究が進められているものの、微量試料からの検討は行われていない。
本研究では、熱安定性に優れた人工核酸Locked Nucleic Acid（LNA）を導入したメチル化解析法を開発することで、微量試料からの高感度な年齢推定法を確立し、遺留資料から被疑者を絞り込む指標としての応用を目指す。

Outline of Final Research Achievements

In this study, we developed the forensic age estimation method of the Japanese population by measuring the methylation rate of DNA using a single-base extension reaction. In addition, we succeeded in increasing the sensitivity by introducing Locked Nucleic Acid （LNA) into each primers.

Academic Significance and Societal Importance of the Research Achievements

本研究成果は、微量な試料からのDNAメチル化率に基づく年齢推定法として法医実務へ利用可能である。また、次世代シーケンサーやパイロシーケンサーを必要とせず、コストパフォーマンスに優れた方法でありその有用性は高い。

Research Products

• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Journal Article] Soluble thrombomodulin ameliorates aberrant hemostasis after rewarming in a rat accidental hypothermia model (2022)
  • Author(s): Takauji Shuhei、Tanaka Hiroki、Hayakawa Mineji、Horioka Kie、Isozaki Shotaro、Konishi Hiroaki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 587 Pages: 1-8
  • DOI: 10.1016/j.bbrc.2021.11.086
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hypoxia-induced nuclear translocation of β-catenin in the healing process of frostbite (2022)
  • Author(s): Isozaki Shotaro、Tanaka Hiroki、Horioka Kie、Konishi Hiroaki、Kashima Shin、Takauji Shuhei、Fujiya Mikihiro、Druid Henrik
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease Volume: 1868 Issue: 6 Pages: 166385-166385
  • DOI: 10.1016/j.bbadis.2022.166385
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The Optimal Dose of Tacrolimus in Combination Therapy with an Anti-TNFα Antibody in a Mouse Colitis Model (2021)
  • Author(s): Murakami Yuki、Fujiya Mikihiro、Konishi Hiroaki、Isozaki Shotaro、Sugiyama Yuya、Kobayashi Yu、Sasaki Takahiro、Kunogi Takehito、Takahashi Keitaro、Ando Katsuyoshi、Ueno Nobuhiro、Kashima Shin、Moriichi Kentaro、Tanabe Hiroki、Okumura Toshikatsu
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 44 Issue: 4 Pages: 564-570
  • DOI: 10.1248/bpb.b20-00916
  • NAID: 130008014301
  • ISSN: 0918-6158, 1347-5215
  • Year and Date: 2021-04-01
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Probiotic Aspergillus oryzae produces anti-tumor mediator and exerts anti-tumor effects in pancreatic cancer through the p38 MAPK signaling pathway. (2021)
  • Author(s): Konishi H, Isozaki S, Kashima S, Moriichi K, Ichikawa S, Yamamoto K, Yamamura C, Ando K, Ueno N, Akutsu H, Ogawa N, Fujiya M.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 11070-11070
  • DOI: 10.1038/s41598-021-90707-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Treatment of hepatocellular carcinoma with autologous platelets encapsulating sorafenib or lenvatinib: A novel therapy exploiting tumor‐platelet interactions (2021)
  • Author(s): Tanaka Hiroki、Horioka Kie、Hasebe Takumu、Sawada Koji、Nakajima Shunsuke、Konishi Hiroaki、Isozaki Shotaro、Goto Masanori、Fujii Yumiko、Kamikokura Yuki、Ogawa Katsuhiro、Nishikawa Yuji
  • Journal Title: International Journal of Cancer Volume: 150 Issue: 10 Pages: 1640-1653
  • DOI: 10.1002/ijc.33915
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Polyphosphate, derived from the probiotic Lactobacillus brevis, modulates the intestinal microbiome and attenuates acute pancreatitis. (2021)
  • Author(s): Takauji S, Konishi H, Fujiya M, Ueno N, Tanaka H, Sato H, Isozaki S, Kashima S, Moriichi K, Mizukami Y, Okumura T.
  • Journal Title: Dig Dis Sci Volume: - Issue: 11 Pages: 1-1
  • DOI: 10.1007/s10620-020-06747-9
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Probiotic derived polyphosphate accelerates intestinal epithelia wound healing through inducing platelet-derived mediators. (2021)
  • Author(s): Isozaki S, Konishi H, Fujiya M, Tanaka H, Murakami Y, Kashima S, Ando K, Ueno N, Moriichi K, Okumura T.
  • Journal Title: Mediators of Inflammation Volume: 2021:5582943 Pages: 1-14
  • DOI: 10.1155/2021/5582943
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Low temperature induces von-willebrand factor expression via increased early growth response 1 transcriptional activity in splenic sinusoidal endothelial cells. (2020)
  • Author(s): Horioka K, Tanaka H, Isozaki S, Konishi H, Addo L, Takauji S, Druid H.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 526 Issue: 1 Pages: 239-245
  • DOI: 10.1016/j.bbrc.2020.03.073
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rewarming from accidental hypothermia enhances whole blood clotting properties in a murine model (2020)
  • Author(s): Horioka Kie、Tanaka Hiroki、Isozaki Shotaro、Konishi Hiroaki、Addo Lynda、Takauji Shuhei、Druid Henrik
  • Journal Title: Thrombosis Research Volume: 195 Pages: 114-119
  • DOI: 10.1016/j.thromres.2020.07.022
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 人工核酸導入Primerを用いた高感度メチル化解析と法医学的年齢推定 (2021)
  • Author(s): 磯崎翔太郎
  • Organizer: 日本DNA多型学会第30回学術集会
• [Presentation] The assessment of cutaneous epithelium hypoxia in the murine frostbite model (2021)
  • Author(s): Shotaro Isozaki, Hiroki Tanaka, Kie Horioka, Hiroaki Konishi, Shin Kashima, Henrik Druid
  • Organizer: 第105次日本法医学会学術全国集会
• [Presentation] 乳酸菌由来長鎖ポリリン酸は血小板活性化を介して腸管上皮の創傷治癒を促進する (2020)
  • Author(s): 磯崎翔太郎、藤谷幹浩、小西弘晃、田中宏樹、村上雄紀、杉山雄哉、 安藤勝祥、上野伸展、後藤拓磨、嘉島 伸、高氏修平、盛一健太郎、 田邊裕貴、水上裕輔、奥村利勝
  • Organizer: 第57回日本消化器免疫学会総会
• [Presentation] The activation of Wnt signaling in the murine frostbite model (2020)
  • Author(s): Shotaro Isozaki, Hiroki Tanaka, Kie Horioka, Hiroaki Konishi, Shin Kashima, Henrik Druid
  • Organizer: 第104次日本法医学会学術全国集会
• [Presentation] PROBIOTIC-DERIVED POLYPHOSPHATE IMPROVES DAMAGED INTESTINAL EPITHELIA THROUGH INDUCING PLATELET-DERIVED CELL GROWTH MOLECULES (2020)
  • Author(s): Shotaro Isozaki , Mikihiro Fujiya , Hiroaki Konishi et al.
  • Organizer: Digestive Disease Week 2020
  • Int'l Joint Research
• [Presentation] Primerへの人工核酸LNA導入によるCpG Islandにおける増幅効率改善効果の検討 (2019)
  • Author(s): 磯崎翔太郎, 小西弘晃
  • Organizer: 第88回日本法医学会学術関東地方集会
• [Remarks] 研究者詳細（磯崎翔太郎）
  • URL: http://hyouka04.asahikawa-med.ac.jp/html/100000707_ja.html
• [Patent(Industrial Property Rights)] 血小板凝集剤 (2020)
  • Inventor(s): 藤谷幹浩,小西弘晃,田中宏樹,磯崎翔太郎,大町佳宏,尾川直樹
  • Industrial Property Rights Holder: 藤谷幹浩,小西弘晃,田中宏樹,磯崎翔太郎,大町佳宏,尾川直樹
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2020-001319
  • Filing Date: 2020