Development of basic technology for intractable and rare diseases

• Project/Area Number: 17K19665
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: General internal medicine and related fields
• Research Institution: Yokohama City University
• Principal Investigator: Akio Yamashita 横浜市立大学, 医学部, 准教授 (20405020)
• Research Collaborator: Usuki Fusako
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: nonsense mutation / mRNA decay / translation / small compound / screening / NMD / 薬剤スクリーニングシステム / 嚢胞性線維症 / モデルマウス / ナンセンス変異 / 希少疾患 / 遺伝性疾患 / リードスルー / 疾患モデルマウス / 転写後制御

Outline of Final Research Achievements

NMD is an mRNA surveillance mechanism that eliminates aberrant mRNAs carrying premature termination codons (PTCs). NMD degrades not only proteins that show dominant-negative function but also aberrant proteins that retain at least some aspect of their normal cell function. If mutant proteins are still functional, the selective inhibition of NMD provides a strategy to ameliorate disease phenotypes in patients with PTC-related conditions. Another strategy to inhibit NMD for the rescue of intractable diseases involves drugs causing translational readthrough. In this project we developed accurate reporter system of NMD and translational readthroug. Using this system, we demonstrated that translational readthrough has little effect on PTC-mRNA decay, NMD inhibition resulted in a synergistic effect on read-through efficiency. We also developed the immortalized cells from Ullrich disease patient and CF model mice with PTC mutation.

Academic Significance and Societal Importance of the Research Achievements

本研究により、異常終止コドンを生じる遺伝子変異による遺伝性疾患について、NMDを抑制することにより治療するための薬剤開発基盤技術創出に成功した。具体的には、薬剤スクリーニングのための改良型高精度NMD検出技術、in vivo POC取得のための、遺伝性疾患患者由来不死化細胞、嚢胞性線維症PTCマウスである。これらの技術を基に得られる薬剤により、これまで治療法のなかった遺伝性疾患について、異常終止コドンまでのタンパク質が機能を有するものについては治療が可能となることが期待される。また、NMD阻害剤とリードスルー薬を組み合わせることにより、多くの遺伝性疾患についても治療可能となることも期待される。

Research Products

• [Journal Article] Effects of rikkunshito on renal fibrosis and inflammation in angiotensin II-infused mice (2019)
  • Author(s): Azushima Kengo、Uneda Kazushi、Wakui Hiromichi、Ohki Kohji、Haruhara Kotaro、Kobayashi Ryu、Haku Sona、Kinguchi Sho、Yamaji Takahiro、Minegishi Shintaro、Ishigami Tomoaki、Yamashita Akio、Tamura Kouichi
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 6201-6201
  • DOI: 10.1038/s41598-019-42657-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effects of ATRAP in Renal Proximal Tubules on Angiotensin‐Dependent Hypertension (2019)
  • Author(s): Kinguchi Sho、Wakui Hiromichi、Azushima Kengo、Haruhara Kotaro、Koguchi Tomoyuki、Ohki Kohji、Uneda Kazushi、Matsuda Miyuki、Haku Sona、Yamaji Takahiro、Yamada Takayuki、Kobayashi Ryu、Minegishi Shintaro、Ishigami Tomoaki、Yamashita Akio、Fujikawa Tetsuya、Tamura Kouichi
  • Journal Title: Journal of the American Heart Association Volume: 8 Issue: 8
  • DOI: 10.1161/jaha.119.012395
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Environmental stresses suppress nonsense-mediated mRNA decay (NMD) and affect cells by stabilizing NMD-targeted gene expression (2019)
  • Author(s): Usuki Fusako、Yamashita Akio、Fujimura Masatake
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 1279-1279
  • DOI: 10.1038/s41598-018-38015-2
  • Peer Reviewed / Open Access
• [Journal Article] Early Enhanced Leucine-Rich α-2-Glycoprotein-1 Expression in Glomerular Endothelial Cells of Type 2 Diabetic Nephropathy Model Mice (2018)
  • Author(s): haku Sona、Wakui Hiromichi、Azushima Kengo、Haruhara Kotaro、Kinguchi Sho、Ohki Kohji、Uneda Kazushi、Kobayashi Ryu、Matsuda Miyuki、Yamaji Takahiro、Yamada Takayuki、Minegishi Shintaro、Ishigami Tomoaki、Yamashita Akio、Ohashi Kenichi、Tamura Kouichi
  • Journal Title: BioMed Research International Volume: 2018 Pages: 1-9
  • DOI: 10.1155/2018/2817045
  • Peer Reviewed / Open Access
• [Journal Article] Angiotensin II Type 1 Receptor-associated Protein Inhibits Angiotensin II-induced Insulin Resistance with Suppression of Oxidative Stress in Skeletal Muscle Tissue (2018)
  • Author(s): Ohki Kohji、Wakui Hiromichi、Kishio Nozomu、Azushima Kengo、Uneda Kazushi、Haku Sona、Kobayashi Ryu、Haruhara Kotaro、Kinguchi Sho、Yamaji Takahiro、Yamada Takayuki、Minegishi Shintaro、Ishigami Tomoaki、Toya Yoshiyuki、Yamashita Akio、Imajo Kento、Nakajima Atsushi、Kato Ikuma、Ohashi Kenichi、Tamura Kouichi
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 2846-2846
  • DOI: 10.1038/s41598-018-21270-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Angiotensin receptor-binding molecule in leukocytes in association with the systemic and leukocyte inflammatory profile (2018)
  • Author(s): Haruhara Kotaro、Wakui Hiromichi、Azushima Kengo、Kurotaki Daisuke、Kawase Wataru、Uneda Kazushi、Haku Sona、Kobayashi Ryu、Ohki Kohji、Kinguchi Sho、Ohsawa Masato、Minegishi Shintaro、Ishigami Tomoaki、Matsuda Miyuki、Yamashita Akio、Nakajima Hideaki、Tamura Tomohiko、Tsuboi Nobuo、Yokoo Takashi、Tamura Kouichi
  • Journal Title: Atherosclerosis Volume: 269 Pages: 236-244
  • DOI: 10.1016/j.atherosclerosis.2018.01.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Endoplasmic reticulum stress preconditioning modifies intracellular mercury content by upregulating membrane transporters (2017)
  • Author(s): Usuki Fusako、Fujimura Masatake、Yamashita Akio
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 12390-12390
  • DOI: 10.1038/s41598-017-09435-3
  • Peer Reviewed / Open Access
• [Journal Article] Angiotensin II Type 1 Receptor‐Associated Protein Regulates Kidney Aging and Lifespan Independent of Angiotensin (2017)
  • Author(s): Uneda Kazushi、Wakui Hiromichi、Maeda Akinobu、Azushima Kengo、Kobayashi Ryu、Haku Sona、Ohki Kohji、Haruhara Kotaro、Kinguchi Sho、Matsuda Miyuki、Ohsawa Masato、Minegishi Shintaro、Ishigami Tomoaki、Toya Yoshiyuki、Atobe Yoshitoshi、Yamashita Akio、Umemura Satoshi、Tamura Kouichi
  • Journal Title: Journal of the American Heart Association Volume: 6 Issue: 8
  • DOI: 10.1161/jaha.117.006120
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ATRAP Expression in Brown Adipose Tissue Does Not Influence the Development of Diet-Induced Metabolic Disorders in Mice (2017)
  • Author(s): Wakui H, Ohki K, Azushima K, Uneda K, Haku S, Kobayashi R, Haruhara K, Kinguchi S, Matsuda M, Ohsawa M, Maeda A, Minegishi S, Ishigami T, Toya Y, Yamashita A, Umemura S, Tamura K
  • Journal Title: International Journal of Molecular Sciences Volume: 18 Issue: 3 Pages: 676-676
  • DOI: 10.3390/ijms18030676
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Translational control of mRNAs by 3'-Untranslated region binding proteins (2017)
  • Author(s): Yamashita Akio、Takeuchi Osamu
  • Journal Title: BMB Reports Volume: 50 Issue: 4 Pages: 194-200
  • DOI: 10.5483/bmbrep.2017.50.4.040
  • Peer Reviewed / Open Access
• [Journal Article] Adipocyte-Specific Enhancement of Angiotensin II Type 1 Receptor-Associated Protein Ameliorates Diet-Induced Visceral Obesity and Insulin Resistance. (2017)
  • Author(s): Azushima K, Ohki K, Wakui H, Uneda K, Haku S, Kobayashi R, Haruhara K, Kinguchi S, Matsuda M, Maeda A, Toya Y, Yamashita A, Umemura S, Tamura K.
  • Journal Title: J Am Heart Assoc. Volume: 6 Issue: 3
  • DOI: 10.1161/jaha.116.004488
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An angiotensin II type 1 receptor binding molecule has a critical role in hypertension in a chronic kidney disease model. (2017)
  • Author(s): Kobayashi R, Wakui H, Azushima K, Uneda K, Haku S, Ohki K, Haruhara K, Kinguchi S, Matsuda M, Ohsawa M, Toya Y, Nishiyama A, Yamashita A, Tanabe K, Maeshima Y, Umemura S, Tamura K.
  • Journal Title: Kidney Int. Volume: 91 Issue: 5 Pages: 1115-1125
  • DOI: 10.1016/j.kint.2016.10.035
  • Peer Reviewed
• [Presentation] PI3 kinase様タンパク質リン酸化酵素SMG1によるアミノ酸飢餓応答依存的遺伝子発現制御 (2018)
  • Author(s): 山下 暁朗
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] PI3 kinase様タンパク質リン酸化酵素SMG1による酸化ストレス応答制御：SMG1によるNRF2活性制御機構の解析 (2018)
  • Author(s): 藤川 由美子, 山下 暁朗, 大貫 哲男, 鈴木 香絵, 青柳 杏子, 黒澤 瞳, 廣瀬 博子, 永井 陽子, 上村 博司, 吉田 稔, 髙橋 秀尚, 大野 茂男
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] SMG1によるNMDとストレス応答統御機構の解析 (2018)
  • Author(s): 山下 暁朗
  • Organizer: 第7回さきがけ「RNAと生体機能」研究会
• [Presentation] PI3-kinase related kinase SMG1 and DNA-PKcs directly phosphorylate and regulate NRF2 in response to oxidative stress. (2018)
  • Author(s): 山下 暁朗
  • Organizer: 京都大タンパク質動態研究所・新学術領域「新生鎖の生物学」合同国際シンポジウム Proteins : from the Cradle to the Grave
• [Presentation] PI3 kinase様タンパク質リン酸化酵素SMG1による新たな転写後制御機構の発見 (2017)
  • Author(s): 山下 暁朗, 佐藤 由典, 黒澤 瞳, 廣瀬 博子, 青柳 杏子, 永井 陽子, 大野 茂男
  • Organizer: 生命科学系学会合同年次大会
• [Presentation] PI3 kinase様タンパク質リン酸化酵素SMG1阻害による抗腫瘍効果の解析 (2017)
  • Author(s): 藤川 由美子, 山下 暁朗, 大貫 哲男, 鈴木 香絵, 青柳 杏子, 黒澤 瞳, 廣瀬 博子, 永井 陽子, 上村 博司, 吉田 稔, 大野 茂男
  • Organizer: 生命科学系学会合同年次大会
• [Presentation] 新規aPKC結合タンパクp200による上皮細胞極性の調節機構 (2017)
  • Author(s): 佐々木 和教, 麹谷 典子, 廣瀬 博子, 吉濱 陽平, 高柳 亜由美, 山下 暁朗, 平野 久, 大野 茂男
  • Organizer: 生命科学系学会合同年次大会