Detection of pancreatic beta cell DNA in the circulation utilizing epigenetic modification

• Project/Area Number: 15K06910
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Medical genome science
• Research Institution: The University of Tokushima
• Principal Investigator: KURODA Akio 徳島大学, 先端酵素学研究所(糖尿病), 准教授 (70571412)
• Co-Investigator(Kenkyū-buntansha): 松久 宗英 徳島大学, 先端酵素学研究所(糖尿病), 教授 (60362737) ; 田蒔 基行 徳島大学, 先端酵素学研究所(糖尿病), 徳島大学専門研究員 (60624400)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: インスリン / 遊離DNA / PCR / bisulfite / 膵β細胞 / インスリン遺伝子 / 1型糖尿病 / DNAメチレーション / ARMS PCR / エピジェネティック制御 / bisulfite処理 / ARMS法 / 制限酵素

Outline of Final Research Achievements

Genomic DNA sequences of any cells are all the same within a body. Pancreatic beta cells only produce insulin and CpG cytosine in the human insulin gene is uniquely unmethylated in pancreatic beta cells. We have developed a new PCR method, which amplifies only if 4 CpG sites in insulin genes were all unmethylated to detect pancreatic beta cell death in the circulation. There were positive results in 18/60 patients with type 1 diabetes and 2/10 patients with type 2 diabetes and 8/21 normal healthy control subjects. This method is able to be applied for the early diagnosis of type 1 diabetes and the detection of transplanted islet or pancreas tissue injury.

Research Products

• [Int'l Joint Research] City of Hope National Medical Center(米国)
• [Presentation] 膵β細胞特異的PCRによる膵β細胞死の検出 (2018)
  • Author(s): 黒田暁生、山田美鈴、冨永ゆかり、鈴木麗子、田蒔基行、明比祐子、高士祐一、石津将、古賀大輔、井本逸勢、倉橋清衛、吉田守美子、遠藤逸朗、粟飯原賢一、安倍正博、Kevin Ferreri、松久宗英
  • Organizer: 第61回日本糖尿病学会年次学術集会
• [Presentation] Detection of pancreatic beta cell DNA in the circulation using the dual Amplification Refractory Mutation System PCR (2018)
  • Author(s): Akio Kuroda, Misuzu Yamashita-Yamada, Yukari Tominaga, Reiko Suzuki, Motoyuki Tamaki, Yuko Akehi, Yuichi Takashi, Daisuke Koga, Eisuke Shimokita, Fuminori Tanihara, Kiyoe Kurahashi, Sumiko Yoshida, Itsuro Endo, Ken-ichi Aihara, Masahiro Abe, Kevin Ferreri, Munehide Matsuhisa
  • Organizer: 78th American Diabetes Association SCIENTIFIC SESSIONS
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] 膵β細胞の傷害検査方法 (2018)
  • Inventor(s): 黒田暁生、山田美鈴、松久宗英
  • Industrial Property Rights Holder: 徳島大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018