Molecular identification of amino acid sensor and its function in mTORC1 signaling pathway

• Project/Area Number: 15K08273
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Osaka University
• Principal Investigator: Ohgaki Ryuichi 大阪大学, 医学系研究科, 助教 (20467525)
• Co-Investigator(Kenkyū-buntansha): 中込 咲綾 大阪大学, 医学(系)研究科(研究院), その他 (60423894)
• Co-Investigator(Renkei-kenkyūsha): OKUDA SUGURU 大阪大学, 大学院医学系研究科, 助教 (50511846) ; NAGAMORI SHUSHI 大阪大学, 大学院医学系研究科, 准教授 (90467572)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: アミノ酸 / mTORC1 / シグナル伝達 / 細胞内シグナル伝達 / 受容体

Outline of Final Research Achievements

Amino acids play an important role as signaling molecules to activate mTORC1, a serine/threonine kinase complex which regulates cellular metabolism. However, the detailed molecular mechanisms of amino acid sensing still remain elusive. In this study, we have tried to understand the molecular basis of amino acids sensing in the upstream of mTORC1, especially focusing on how the cells recognize the extracellular amino acids. The experimental protocol for the comparative proteomics analysis using the plasma membrane fractions has been established. Also, the conditions of cell treatments as well as the methods for detection of mTORC1 activity were optimized, to improve the reliability and the throughput of the analyses. The outcomes the research would significantly contribute to identifying the amino acid sensing molecules in the future studies.

Research Products

• [Journal Article] Essential roles of L-type amino acid transporter 1 in syncytiotrophoblast development by presenting fusogenic 4F2hc. (2017)
  • Author(s): Ohgaki R, Ohmori T, Hara S, Nakagomi S, Kanai-Azuma M, Kaneda-Nakashima K, Okuda S, Nagamori S, Kanai Y.
  • Journal Title: Mol Cell Biol. Volume: 印刷中 Issue: 11
  • DOI: 10.1128/mcb.00427-16
  • Peer Reviewed / Open Access
• [Journal Article] Ratiometric fluorescence imaging of cell surface pH by poly(ethylene glycol)-phospholipid conjugated with fluorescein isothiocyanate (2017)
  • Author(s): Ryuichi Ohgaki, Yuji Teramura, Daichi Hayashi, Suguru Okuda, Shushi Nagamori, Madoka Taka and Yoshikatsu Kanai
  • Journal Title: Sci Res Volume: 7 Issue: 1 Pages: 17484-17484
  • DOI: 10.1038/s41598-017-17459-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure-activity relationship of a novel series of inhibitors for cancer type transporter L-type amino acid transporter 1 (LAT1). (2017)
  • Author(s): Kongpracha P, Nagamori S, Wiriyasermkul P, Tanaka Y, Kaneda K, Okuda S, Ohgaki R, Kanai Y.
  • Journal Title: J Pharmacol Sci. Volume: 133 Issue: 2 Pages: 96-102
  • DOI: 10.1016/j.jphs.2017.01.006
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Interaction of the Sodium/Glucose Cotransporter (SGLT) 2 inhibitor Canagliflozin with SGLT1 and SGLT2: Inhibition kinetics, sidedness of action, and transporter-associated incorporation accounting for its pharmacodynamic and pharmacokinetic features. (2016)
  • Author(s): Ohgaki R, Wei L, Yamada K, Hara T, Kuriyama C, Okuda S, Ueta K, Shiotani M, Nagamori S, Kanai Y.
  • Journal Title: J Pharmacol Exp Ther. Volume: 358 Issue: 1 Pages: 94-102
  • DOI: 10.1124/jpet.116.232025
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Impaired Amino Acid Transport at the Blood Brain Barrier Is a Cause of Autism Spectrum Disorder. (2016)
  • Author(s): Tarlungeanu DC, Deliu E, Dotter CP, Kara M, Janiesch PC, Scalise M, Galluccio M, Tesulov M, Morelli E, Sonmez FM, Bilguvar K, Ohgaki R, Kanai Y, Johansen A, Esharif S, Ben-Omran T, Topcu M, Schlessinger A, Indiveri C, Duncan KE, Caglayan AO, Gunel M, Gleeson JG, Novarino G.
  • Journal Title: Cell. Volume: 167 Issue: 6 Pages: 1481-1494
  • DOI: 10.1016/j.cell.2016.11.013
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transport of 3-fluoro-l-α-methyl-tyrosine (FAMT) by organic ion transporters explains renal background in [(18)F]FAMT positron emission tomography. (2016)
  • Author(s): Wei L, Tominaga H, Ohgaki R, Wiriyasermkul P, Hagiwara K, Okuda S, Kaira K, Kato Y, Oriuchi N, Nagamori S, Kanai Y.
  • Journal Title: J Pharmacol Sci. Volume: 130 Issue: 2 Pages: 101-109
  • DOI: 10.1016/j.jphs.2016.01.001
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Specific transport of 3-fluoro-l-α-methyl-tyrosine by LAT1 explains its specificity to malignant tumors in imaging. (2016)
  • Author(s): Wei L, Tominaga H, Ohgaki R, Wiriyasermkul P, Hagiwara K, Okuda S, Kaira K, Oriuchi N, Nagamori S, Kanai Y.
  • Journal Title: Cancer Sci. Volume: 107 Issue: 3 Pages: 347-352
  • DOI: 10.1111/cas.12878
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Novel cystine transporter in renal proximal tubule identified as a missing partner of cystinuria-related plasma membrane protein rBAT/SLC3A1. (2016)
  • Author(s): Nagamori S, Wiriyasermkul P, Guarch ME, Okuyama H, Nakagomi S, Tadagaki K, Nishinaka Y, Bodoy S, Takafuji K, Okuda S, Kurokawa J, Ohgaki R, Nunes V, Palacín M, Kanai Y.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 113 Issue: 3 Pages: 775-780
  • DOI: 10.1073/pnas.1519959113
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Structure-activity relations of leucine derivatives reveal critical moieties for cellular uptake and activation of mTORC1-mediated signaling. (2016)
  • Author(s): Nagamori S, Wiriyasermkul P, Okuda S, Kojima N, Hari Y, Kiyonaka S, Mori Y, Tominaga H, Ohgaki R, Kanai Y.
  • Journal Title: Amino Acids. Volume: 48 Issue: 4 Pages: 279-286
  • DOI: 10.1007/s00726-015-2158-z
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 腫瘍血管新生に寄与する血管内皮細胞のアミノ酸トランスポーター (2018)
  • Author(s): 大垣隆一、Quan Lili、奥田傑、永森收志、金井好克
  • Organizer: 第1回 がんと代謝研究会・若手の会
• [Presentation] SGLT2阻害薬カナグリフロジンのヒトSGLT1およびSGLT2に対する作用キネティクスと阻害メカニズムの検討 (2017)
  • Author(s): 大垣隆一、Ling Wei、山田和徳、原大樹、栗山千亜紀、奥田傑、植田喜一郎、Laurent Leclercq、Rao N.V.S. Mamidi、塩谷正治、永森收志、金井好克
  • Organizer: 第90回 日本薬理学会年会
  • Place of Presentation: 長崎
  • Year and Date: 2017-03-15
• [Presentation] アミノ酸トランスポーターLAT1によるロイシン誘導体の認識とmTORC1シグナル活性化の関係 (2017)
  • Author(s): 奥田傑、Pattama Wiriyasermkul、大垣隆一、永森收志、金井好克
  • Organizer: 第90回 日本薬理学会年会
  • Place of Presentation: 長崎
  • Year and Date: 2017-03-15
• [Presentation] 有機アニオントランスポーターOAT1に対する芳香族アミノ酸誘導体の構造活性相関：イメージングプローブの腎臓におけるバックグラウンド集積を誘起する分子内構造の同定 (2017)
  • Author(s): 魏玲、大垣隆一、富永英之、兼田加珠子、奥田傑、永森收志、金井好克
  • Organizer: 第90回 日本薬理学会年会
  • Place of Presentation: 長崎
  • Year and Date: 2017-03-15
• [Presentation] A C-terminus mutation of SLC7A9 amino acid transporter retains substrate binding but diminishes the transport activity. (2017)
  • Author(s): Nagamori S, Kawamoto Y, Wiriyasermkul P, Nakagomi S, Okuda S, Ohgaki R, Kanai Y.
  • Organizer: Gordon Research Conference: Mechanisms of Membrane Transport
  • Int'l Joint Research
• [Presentation] Structure-activity relationship of novel inhibitors for cancer type amino acid transporter LAT1 (2017)
  • Author(s): Pornparn Kongpracha, Pattama Wiriyasermkul,Kazuko Kaneda-Nakashima, Suguru Okuda, Ryuichi Ohgaki, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 第131回 日本薬理学会近畿部会
• [Presentation] アミノ酸トランスポーターLAT1は膜融合関連因子4F2hcの細胞膜提示による胎盤合胞体性栄養膜の形成に必須である (2017)
  • Author(s): 大垣隆一、大森崇弘、原早織 、中込咲綾 、金井正美 、兼田（中島）加珠子、奥田傑、○永森收志、金井好克
  • Organizer: 第12回 トランスポーター研究会年会
• [Presentation] Phosphoproteomics reveals the cellular processes affected by leucine transported through LAT1, a cancer type amino acid transporter”. (2017)
  • Author(s): Pornparn Kongpracha, Pattama Wiriyasermkul,, Suguru Okuda, Ryuichi Ohgaki, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 日本プロテオーム学会2017年大会 JHUPO 第15回大会
• [Presentation] Critical moieties of aromatic amino acid for renal accumulation of tumor imaging probes (2017)
  • Author(s): Chunhuan Jin, Ling Wei, Ryuichi Ohgaki, Hideyuki Tominaga, Suguru Okuda, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 第132回 日本薬理学会近畿部会
• [Presentation] LAT1 in endothelial cells as a novel target of anti-angiogenic cancer therapy (2017)
  • Author(s): Lili Quan, Ryuichi Ohgaki, Suguru Okuda, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 第132回 日本薬理学会近畿部会
• [Presentation] Non-competitive LAT1 inhibitors suppress mTORC1 signaling similary to competitive LAT1 inhibitors (2017)
  • Author(s): Pornparn Kongpracha, Suguru Okuda, Ryuichi Ohgaki, Yoshikatsu Kanai, Shushi Nagamori
  • Organizer: 第132回 日本薬理学会近畿部会
• [Presentation] アミノ酸トランスポーターLAT1は胎盤合胞体性栄養膜の形成に必須である：膜融合関連因子4F2hcの細胞膜提示による細胞融合への寄与 (2016)
  • Author(s): 大垣 隆一、大森 崇弘、原 早織、中込 咲綾、金井 正美、兼田 加珠子、奥田 傑、永森 收志、金井 好克
  • Organizer: 第39回 日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] PEG結合リン脂質型蛍光分子プローブの 細胞膜表層pHイメージングツールとしての有用性の検討 (2016)
  • Author(s): 大垣 隆一、寺村 裕治、林 大智、永森 收志、高井 まどか、金井 好克
  • Organizer: H28年度 生理研研究会（第5回 生体界面研究会
  • Place of Presentation: 岡崎
  • Year and Date: 2016-07-04
• [Presentation] Inhibition kinetics, sidedness of action, and transporter-mediated uptake of the sodium/glucose cotransporter (SGLT) 2 inhibitor canagliflozin: implications for its pharmacodynamic and pharmacokinetic features (2016)
  • Author(s): Ohgaki R, Wei L, Yamada K, Hara T, Kuriyama C, Okuda S, Ueta K, Leclercq L, Mamidi RN, Shiotani M, Nagamori S, Kanai Y
  • Organizer: 76th American Diabetes Association Scientific Sessions
  • Place of Presentation: New Orleans, Louisiana, US
  • Year and Date: 2016-06-10
  • Int'l Joint Research
• [Presentation] Involvements of LRRC8A and amino acid transporter SLC proteins in the activity of volume-sensitive outwardly rectifying anion channel (VSOR) (2016)
  • Author(s): Toshiaki Okada, Ravshan Z. Sabirov, Ryuichi Ohgaki, Shushi Nagamori, Yoshikatsu Kanai, Koh Ono, Hiromi Uramoto, Yasunobu Okada
  • Organizer: 第93回 日本生理学会大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-03-22
• [Presentation] Interaction of potential hepatotoxic ingredients of Traditional Chinese Medicine with liver drug transporters OATP1B1 and MDR1 (2016)
  • Author(s): Lili Quan，Zijie Song, Yueqing Zhou, Ryuichi Ohgaki, Shushi Nagamori, Xin He, Yoshikatsu Kanai
  • Organizer: 第63回 日本生化学会 近畿支部例会学会
  • Place of Presentation: 神戸
• [Presentation] Interaction of halogenated aromatic amino acid probes for tumor imaging with renal organic anion transporter OAT1: the mechanisms of renal accumulation of the imaging probes (2016)
  • Author(s): Ling Wei、大垣 隆一、富永 英之、Quan Lili、兼田（中島）加珠子、奥田 傑、永森 收志、金井 好克
  • Organizer: 第129回 日本薬理学会近畿部会
  • Place of Presentation: 広島
• [Presentation] PEG結合リン脂質型蛍光分子プローブによる細胞膜表層pHのイメージング (2016)
  • Author(s): 大垣 隆一、寺村 裕治、林 大智、永森 收志、高井 まどか、金井 好克
  • Organizer: 第129回 日本薬理学会近畿部会
  • Place of Presentation: 広島
• [Presentation] Inhibition kinetics, sidedness of action, and transporter-mediated uptake of the sodium/glucose cotransporter (SGLT) 2 inhibitor canagliflozin: implications for its pharmacodynamic and pharmacokinetic features (2016)
  • Author(s): Ling Wei, Ryuichi Ohgaki, Kazunori Yamada, Taiki Hara, Chiaki Kuriyama, Suguru Okuda, Kiichiro Ueta, Masaharu shiotani, Shushi Nagamori and Yoshikatsu Kanai
  • Organizer: International and Interdisciplinary Symposium 2016 “Towards a New Era of Cardiovascular Research”
  • Place of Presentation: 東京
• [Presentation] アミノ酸トランスポーターLAT1による膜融合関連因子4F2hcの細胞膜提示は胎盤合胞体性栄養膜の形成に必須である (2016)
  • Author(s): 大垣隆一
  • Organizer: 大阪大学医学系研究フォーラム 第7回 若手研究フォーラム
  • Place of Presentation: 大阪
• [Presentation] アミノ酸トランスポーターLAT1による基質認識とmTORC1シグナル活性化に必要なロイシンの構造部位の決定 (2016)
  • Author(s): 奥田 傑, Pattama Wiriyasermkul, 大垣 隆一, 永森 收志, 金井 好克
  • Organizer: 第130回 日本薬理学会近畿部会
  • Place of Presentation: 京都
• [Presentation] アミノ酸トランスポーターLAT1阻害によるがん転移抑制効果の検討 (2016)
  • Author(s): 兼田 加珠子, 魏 玲, Pornparn Kongpracha, 大垣 隆一, 永森 收志, 金井 好克
  • Organizer: 第130回 日本薬理学会近畿部会
  • Place of Presentation: 京都
• [Presentation] Inhibitor of amino acid transporter is useful as a novel anti-cancer drug (2015)
  • Author(s): 兼田（中島）加珠子、疋田隼人、Pornparn Kongpracha、Ling Wei、大垣隆一、永森收志、竹原徹郎、金井好克
  • Organizer: 第38回 日本分子生物学会年会・第88回 日本生化学会大会 合同大会 (BMB2015)
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Leucine transported by a cancer-type amino acid transporter LAT1 affects multiple cellular processes in pancreatic cancer cells. (2015)
  • Author(s): Pornparn Kongpracha, Pattama Wiriyasermkul, Noriyoshi Isozumi, Kazuko Kaneda-Nakashima, Suguru Okuda, Ryuichi Ohgaki, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 第38回 日本分子生物学会年会・第88回 日本生化学会大会 合同大会 (BMB2015)
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Properties of mTORC1 regulation by amino acid signaling in HEK293T cells (2015)
  • Author(s): 大垣 隆一、永森 收志、ウィリヤサムクン パッタマ、萩原 浩平、金井 好克
  • Organizer: 第38回 日本分子生物学会年会・第88回 日本生化学会大会 合同大会 (BMB2015)
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] アミノ酸トランスポーター阻害薬による抗腫瘍薬作用増強効果についての検討 (2015)
  • Author(s): 兼田加珠子、Pornparn Kongpracha、Ling Wei、大垣 隆一、 永森 收志、金井 好克
  • Organizer: 第37回 生体膜と薬物の相互作用シンポジウム
  • Place of Presentation: 熊本
  • Year and Date: 2015-11-19
• [Presentation] アミノ酸輸送体阻害薬による抗がん剤作用増強効果についての検討 (2015)
  • Author(s): 兼田加珠子、Pornparn Kongpracha、Ling Wei、大垣 隆一、永森 收志、金井 好克
  • Organizer: 第24回 日本Cell Death学会
  • Place of Presentation: 大阪
  • Year and Date: 2015-07-11
• [Presentation] Transport of 3-fluoro-L-α-methyl-tyrosine (FAMT) by organic ion transporters: a possible cause of renal background in 18F-FAMT PET (2015)
  • Author(s): Wei Ling、富永 英之、Wiriyasermkul Pattama、大垣 隆一、萩原 浩平、奥田 傑、永森 收志、金井 好克
  • Organizer: 第127回 日本薬理学会近畿部会
  • Place of Presentation: 岐阜
  • Year and Date: 2015-06-26
• [Presentation] Inhibition of the LAT1-mediated leucine transport affects multiple cellular processes in pancreatic cancer cells. (2015)
  • Author(s): Pornparn Kongprach, Pattama Wiriyasermkul, Noriyoshi Isozumi, Printip Wongthai, Suguru Okuda, Kenjiro Tadagaki, Ryuichi Ohgaki, Shushi Nagamori, Yoshikatsu Kanai
  • Organizer: 第62回 日本生化学会近畿支部例会
  • Place of Presentation: 滋賀
  • Year and Date: 2015-05-16