ミトコンドリア遺伝子異常糖尿病に対する遺伝子治療の開発

• Project/Area Number: 18659263
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Metabolomics
• Research Institution: Tohoku University
• Principal Investigator: 檜尾 好徳 (桧尾 好徳) Tohoku University, 大学院・医学系研究科, 准教授 (10282071)
• Co-Investigator(Kenkyū-buntansha): 石原 寿光 東北大学, 病院, 講師 (60361086)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,300,000 (Direct Cost: ¥3,300,000); Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,300,000); Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: 糖尿病 / ミトコンドリア / ミトコンドリアDNA / 一塩基置換修復法 / Chimeric RNA / DNA / 膵ラ島β細胞 / インスリン / ガングリオシド抗体

Research Abstract

ミトコンドリアはDNA修復酵素系に富み,ダイナミックにミトコンドリアDNA(mtDNA)変異の修復を受けている.mtDNA修復系の特徴から,Chimeric RNA/DNA oligonucleotidesの有効性が期待されている.海外共同研究者のd'Alarcao教授らはoligonucleotidesをアルキルトリフェニルホスホネート(APP)誘導体化すると,ミトコンドリア内へ効率よく集積することを発見した.本研究では,mtDNA変異を標的としたChimeric RNA/DNA oligonucleotidesを作成し,APPで誘導体化して,ミトコンドリア集積性を高めて,mtDNA変異を標的とする一塩基置換修復法の開発を試みた.
1.ミトコンドリアDNAを分子標的とする特異的一塩基置換修復法の開発:ミトコンドリア
DNA3243A/G変異を標的とするChimeric RNA/DNA oligonucleotidesを化学合成した.正常ミトコンドリアDNAと3243G変異DNAのmixtureをテンプレートとしたIn VitroミトコンドリアDNA複製系に,Chimeric RNA/DNA oligonucleotidesを添加すると,容量依存性に3243G変異が修復された.
2.化学合成Chimeric RNA/DNA oligonucleotidesのAPP誘導体化:Chimeric RNA/DNA
oligonucleotidesを化学合成し,アルキルトリフェニルホスホネート誘導体(APP)による修飾が可能か,検討した.対照としたDNA oligonucleotidesに比べて,Chimeric RNA/DNA oligonucleotidesはAPP化修飾効率が対照の約20%と低い.

Research Products

• [Journal Article] A novel method for evaluating human carotid artery elasticity: possible detection of early stage atherosclerosis in subjects with type 2 diabetes. (2008)
  • Author(s): Okimoto H, Ishigaki Y, Koiwa Y, Hinokio Y, Ogihara T, Suzuki S, Katagiri H, Ohkubo T, Hasegawa H,
  • Journal Title: Atherosclerosis. 196 Pages: 391-397
  • Peer Reviewed
• [Journal Article] Possible relevance of HLA-DRB1*0403 haplotype in insulin autoimmune syndrome induced by α-lipoic acid, used as a dietary supplement. (2007)
  • Author(s): Yamada T, Imai J, Ishigaki Y, Hinokio Y, Oka Y, Katagiri H.
  • Journal Title: Diabetes Care 30 Pages: 131-131
  • Peer Reviewed
• [Journal Article] Increased hepatic expression of ganglioside-specific sialidase, NEU3, improves insulin sensitivity and glucose tolerance in mice. (2007)
  • Author(s): Yoshizumi S, Suzuki S, Hirai M, Hinokio Y, Yamada T, Yamada T,
  • Journal Title: Metabolism 56 Pages: 420-429
  • Peer Reviewed
• [Journal Article] Reliability and validity of the Japanese version of the World Health Organization-Five Well-Being Index in the context of detecting depression in diabetic patients. (2007)
  • Author(s): Awata S, Bech P, et al.
  • Journal Title: Psychiatry Clin Neurosci. 61・1 Pages: 112-117
• [Journal Article] Increased hepatic expression of ganglioside-specific sialidase, NEU3, improves insulin sensitivity and glucose tolerance in mice. (2007)
  • Author(s): Yoshizumi S, Suzuki S, et al.
  • Journal Title: Metabolism 56・3 Pages: 420-439
• [Journal Article] A novel method for evaluating human carotid artery elasticity : Possible detection of early stage atherosclerosis in subjects with type 2 diabetes. (2007)
  • Author(s): Okimoto H, Ishigaki Y, et al.
  • Journal Title: Atherosclerosis (印刷中)
• [Journal Article] Cell type-specific activation of metabolism reveals that beta-cell secretion suppresses glucagon release from alpha-cells in rat pancreatic islets. (2006)
  • Author(s): Takahashi R, Ishihara H, et al.
  • Journal Title: Am J Physiol Endocrinol Metab 290・2
• [Journal Article] WFS1 protein modulates the free Ca(2+)concentration in the endoplasmic reticulum. (2006)
  • Author(s): Takei D, Ishihara H, et al.
  • Journal Title: FEBS Lett. 580・24 Pages: 5635-5640
• [Journal Article] WFS1-deficiency increases endoplasmic reticulum stress, impairs cell cycle progression and triggers the apoptotic pathway specifically in pancreatic beta-cells. (2006)
  • Author(s): Yamada T, Ishihara H, et al.
  • Journal Title: Hum Mol Genet. 15・10 Pages: 1600-1609