Identification of mitochondrial aconitase as a critical enzyme sensitive to microgravity stress

• Project Area: "LIVING IN SPACE" - Integral Understanding of life-regulation mechanism from "SPACE"
• Project/Area Number: 16H01645
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: The University of Tokushima
• Principal Investigator: 二川 健 徳島大学, 大学院医歯薬学研究部(医学系), 教授 (20263824)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000); Fiscal Year 2017: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2016: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
• Keywords: 無重力センサー / 酸化ストレス / ミトコンドリア / アコニターゼ / 筋萎縮 / 無重力環境 / メカニカルストレス / メタボローム解析

Outline of Annual Research Achievements

筋萎縮などメカニカルストレスの応答異常による疾患(いわゆる無重力病)は代謝疾患である。つまり、メカニカルストレスに対する臓器・細胞の応答異常は、遺伝子異常を伴わない。研究代表者は、究極のメカニカルストレスである、無重力環境に対する生体の応答を、網羅的な代謝レベルの解析(メタボローム解析)を基盤に研究してきた。本研究では、無重力ストレスに対する筋細胞の初期応答（シグナル・トランスダクション）を明らかにした。具体的には、
1．「酸化ストレスによるミトコンドリア機能異常」：もう一つは、酸化ストレスがミトコンドリア・アコニターゼ活性を不活化し、ミトコンドリアの機能障害を引き起こす経路である。宇宙空間で培養したL6細胞において、TCAサイクル上の中間生成物であるcisアコニット酸の蓄積が見られていたため、これらを触媒する酵素であり、酸化ストレス感受性を持つアコニターゼに注目した。そこで、3D-Clinorotationを行い、アコニターゼ活性について調べたところ、活性の有意な低下が見られた。
2．「アコニターゼノックダウンの筋管細胞への影響」：アコニターゼノックダウンは筋管細胞の好気的解糖系を障害し、嫌気的解糖系を高めた。その結果、ATP産生量は減少した。さらに、アコニターゼノックダウンはERK1/2とDrp1の活性化を介してミトコンドリアの断片化を誘導し、遅筋型のミオシン重鎖の分解を引き起こした。
3．萎縮筋の見られた酸化ストレスの亢進は、ERK1/2-Egr-Cbl-bの発現経路を介して筋量の減少を、Aconitase活性低下－Mitochondrial dysfunctionを介して筋質（遅筋から速筋へ）の変化を誘導していることがわかった。また、抗酸化剤はクリノスタットによるERK1/2の活性化を抑制した。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] VEGF pathway-targeting drugs induce evasive adaptation by activation of neuropilin-1/cMet in colon cancer cells (2018)
  • Author(s): Tomida Chisato、Yamagishi Naoko、Nagano Hikaru、Uchida Takayuki、Ohno Ayako、Hirasaka Katsuya、Nikawa Takeshi、Teshima-Kondo Shigetada
  • Journal Title: International Journal of Oncology Volume: 52 Pages: 1350-1362
  • DOI: 10.3892/ijo.2018.4291
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distinct Gene Expression Profile Distinguishes Increased Metabolic Activity in Spontaneously Hyperactive Rats While Sedentary from That Induced by Exercise (2018)
  • Author(s): Abe Manami、Matsuo Yuki、Harada-Sukeno Akiko、Uchida Takayuki、Kitahata Kanako、Tomida Chisato、Hirasaka Katsuya、Teshima-Kondo Shigetada、Harada Nagakatsu、Nakaya Yutaka、Sakaue Hiroshi、Nakao Reiko、Nikawa Takeshi
  • Journal Title: Advances in Biological Chemistry Volume: 08 Issue: 01 Pages: 1-14
  • DOI: 10.4236/abc.2018.81001
  • Peer Reviewed
• [Journal Article] Reactive oxygen species up-regulate expression of muscle atrophy-associated ubiquitin ligase Cbl-b in rat L6 skeletal muscle cells. (2018)
  • Author(s): Uchida T, Sakashita Y, Kitahata K, Yamashita Y, Tomida C, Kimori Y, Komatsu A, Hirasaka K, Ohno A, Nakao R, Higashitani A, Higashibata A, Ishioka N, Shimazu T, Kobayashi T, Okumura Y, Choi I, Oarada M, Mills EM, Teshima-Kondo S, Takeda S, Tanaka E, Tanaka K, Sokabe M, Nikawa T.
  • Journal Title: Am J Physiol Cell Physiol Volume: 印刷中
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Co-Administration of Myostatin-Targeting siRNA and ActRIIB-Fc Fusion Protein Increases Masseter Muscle Mass and Fiber Size (2017)
  • Author(s): BAYARSAIKHAN Od、KAWAI Nobuhiko、MORI Hiroyo、KINOUCHI Nao、NIKAWA Takeshi、TANAKA Eiji
  • Journal Title: Journal of Nutritional Science and Vitaminology Volume: 63 Issue: 4 Pages: 244-248
  • DOI: 10.3177/jnsv.63.244
  • NAID: 130007314434
  • ISSN: 0301-4800, 1881-7742
  • Peer Reviewed / Open Access
• [Journal Article] Update on recent progress in vitamin D research. Vitamin D and muscle tissues (2017)
  • Author(s): Kaneko I, Miyamoto KI, Nikawa T
  • Journal Title: Clin Calcium Volume: 27 Pages: 1571-1578
• [Journal Article] Regorafenib induces adaptive resistance of colorectal cancer cells via inhibition of vascular endothelial growth factor receptor (2017)
  • Author(s): Tomida Chisato、Nagano Hikaru、Yamagishi Naoko、Uchida Takayuki、Ohno Ayako、Hirasaka Katsuya、Nikawa Takeshi、Teshima-Kondo Shigetada
  • Journal Title: The Journal of Medical Investigation Volume: 64 Issue: 3.4 Pages: 262-265
  • DOI: 10.2152/jmi.64.262
  • NAID: 130006105177
  • ISSN: 1343-1420, 1349-6867
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Antiangiogenic agent sunitinib induces epithelial to mesenchymal transition and accelerates motility of colorectal cancer cells (2017)
  • Author(s): Chisato Tomida, Naoko Yamagishi, Hikaru Nagano, Takayuki Uchida, Ayako Ohno, Katsuya Hirasaka, Takeshi Nikawa, Shigetada Teshima-Kondo
  • Journal Title: The Journal of Medical Investigation Volume: 64 Issue: 3.4 Pages: 250-254
  • DOI: 10.2152/jmi.64.250
  • NAID: 130006105181
  • ISSN: 1343-1420, 1349-6867
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distinct effects of dietary whey peptide and soy protein on denervation-mediated muscle atrophy. (2017)
  • Author(s): Takagi M, Uchida T, Takatsu E, Kishimoto H, Ida K, Ishida Y, Ono-Ohmachi A, Morita Y, Kato K, Ochi A, Ohno-Maita A, Oarada M, Abe T, Teshima-Kondo S, Nakao R, Hirasaka K, Nikawa T*.
  • Journal Title: J Nutr Food Sci Volume: 印刷中
  • Open Access / Int'l Joint Research
• [Journal Article] Modulation of cutaneous extracellular collagen contraction by phosphorylation status of p130Cas (2017)
  • Author(s): Takeya M, Okumura Y, Nikawa T*
  • Journal Title: J Physiol Sci Volume: 印刷中
  • NAID: 40021302104
  • Peer Reviewed
• [Journal Article] Metabolic Suppression by 3-Iodothyronamine Induced Muscle Cell Atrophy <i>via</i> Activation of FoxO–Proteasome Signaling and Downregulation of Akt1–S6K Signaling (2017)
  • Author(s): Ju H, Kim T, Chung CM, Park J, Nikawa T, Park K, Choi I*
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 40 Issue: 5 Pages: 576-582
  • DOI: 10.1248/bpb.b16-00653
  • NAID: 130005631971
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Development and performance evaluation of a three-dimensional clinostat synchronized heavy-ion irradiation system (2017)
  • Author(s): Ikeda H, Hidema J and Takahashi A et al.
  • Journal Title: Life Sci Space Res Volume: 12 Pages: 51-60
  • DOI: 10.1016/j.lssr.2017.01.003
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Nutritional counseling regulates interdialytic weight gain and blood pressure in outpatients receiving maintenance hemodialysis (2017)
  • Author(s): Sakai A, Hamada H, Hara K, Mori K, Uchida T, Mizuguchi T, Minaguchi J, Shima K, Kawashima S, Hamada Y, Nikawa T*.
  • Journal Title: J Med Invest Volume: 64 Pages: 129-135
  • Peer Reviewed
• [Journal Article] Involvement of Cbl-b-mediated macrophage inactivation in insulin resistance. (2017)
  • Author(s): Abe T, Hirasaka K, Nikawa T.
  • Journal Title: World J Diabetes. Volume: 8 Issue: 3 Pages: 97-103
  • DOI: 10.4239/wjd.v8.i3.97
  • NAID: 120006937652
  • Peer Reviewed / Open Access
• [Journal Article] UCP3 is associated with Hax-1 in mitochondria in the presence of calcium ion (2016)
  • Author(s): Hirasaka K, Mills EM, Haruna M, Bando A, Ikeda C, Abe T, Kohno S, Nowinski SM, Lago CU, Akagi K, Tochio H, Ohno A, Teshima-Kondo S, Okumura Y, Nikawa T
  • Journal Title: Biochem Biophys Res Commun Volume: 472 Issue: 1 Pages: 108-113
  • DOI: 10.1016/j.bbrc.2016.02.075
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Capric Acid Up-Regulates UCP3 Expression without PDK4 Induction in Mouse C2C12 Myotubes (2016)
  • Author(s): Abe T, Hirasaka K, Kohno S, Tomida C, Haruna M, Uchida T, Ohno A, Oarada M, Teshima-Kondo S, Okumura Y, Choi I, Aoyama T, Terao J, Nikawa T*.
  • Journal Title: Journal of Nutritional Science and Vitaminology Volume: 62 Issue: 1 Pages: 32-39
  • DOI: 10.3177/jnsv.62.32
  • NAID: 130005148603
  • ISSN: 0301-4800, 1881-7742
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Preventive effect of dietary quercetin on disuse muscle atrophy by targeting mitochondria in denervated mice (2016)
  • Author(s): Mukai R, Matsui N, Fujikura Y, Matsumoto N, Hou DX, Kanzaki N, Shibata H, Horikawa M, Iwasa K, Hirasaka K, Nikawa T, Terao J
  • Journal Title: J Nutr Biochem Volume: 31 Pages: 67-76
  • DOI: 10.1016/j.jnutbio.2016.02.001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Dietary Supplementation with Isoflavones Prevents Muscle Wasting in Tumor-Bearing Mice (2016)
  • Author(s): Hirasaka K*, Saito S, Yamaguchi S, Miyazaki R, Wang Y, Haruna M, Taniyama S, Higashitani A, Terao J, Nikawa T, Tachibana K
  • Journal Title: Journal of Nutritional Science and Vitaminology Volume: 62 Issue: 3 Pages: 178-184
  • DOI: 10.3177/jnsv.62.178
  • NAID: 130005253104
  • ISSN: 0301-4800, 1881-7742
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 8-Prenylnaringenin promotes recovery from immobilization-induced disuse muscle atrophy through activation of the Akt phosphorylation pathway in mice (2016)
  • Author(s): Mukai R, Horikawa H, Lin PY, Tsukumo N, Nikawa T, Kawamura T, Nemoto H, Terao J*
  • Journal Title: Am J Physiol Regul Integr Comp Physiol Volume: 311 Issue: 6 Pages: 1022-1031
  • DOI: 10.1152/ajpregu.00521.2015
  • NAID: 120006535072
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The malignant progression effects of regorafenib in human colon cancer cells (2015)
  • Author(s): Tomida C, Aibara K, Yamagishi N, Yano C, Nagano H, Abe T, Ohno A, Hirasaka K, Nikawa T, Teshima-Kondo S
  • Journal Title: The Journal of Medical Investigation Volume: 62 Issue: 3.4 Pages: 195-198
  • DOI: 10.2152/jmi.62.195
  • NAID: 130005099146
  • ISSN: 1343-1420, 1349-6867
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Molecular mechanism and nutritional approach for unloading-mediated muscle atrophy (2018)
  • Author(s): Takeshi Nikawa, Takayuki Uchida, Ayako Miata (Ohno), Reiko Nakao
  • Organizer: 2018 International Conference on Functional Food for improving Sarcopenia, Cachexia and Frailty
  • Int'l Joint Research / Invited
• [Presentation] 骨格筋蛋白質代謝と骨格筋量調節 (2017)
  • Author(s): 二川健
  • Organizer: 第20回日本病態栄養学会年次学術集会
  • Place of Presentation: 京都国際会議場（京都府・京都市）
  • Year and Date: 2017-01-15
  • Invited
• [Presentation] Unloading induces reactive oxygen species associated signal transduction toward atrophy in skeletal muscle cells (2017)
  • Author(s): Takeshi Nikawa
  • Organizer: 3rd Internation Symposium on Mechanobiology
  • Int'l Joint Research / Invited
• [Presentation] 無重力ストレスに対する初期応答酵素（アコニターゼ）の筋萎縮における役割 (2017)
  • Author(s): 二川健
  • Organizer: 2017年度生命科学系学会合同年次大会第40回日本分子生物学会年会、第90回日本生化学会大会
• [Presentation] Molecular mechanism of microgravity-induced muscle atrophy (2017)
  • Author(s): 二川健
  • Organizer: 第65回国際歯科研究学会日本部会総会・学術大会
  • Invited
• [Presentation] Unloading induces reactive oxygen species associated signal transduction toward atrophy in skeletal muscle cells (2017)
  • Author(s): Takeshi Nikawa
  • Organizer: 14th Japan-Korea Joint Seminar on Space Environment Utilization Research
  • Int'l Joint Research
• [Presentation] 無重力ストレスによる筋萎縮における酸化ストレスの重要性 (2017)
  • Author(s): 内田貴之、真板綾子、中尾玲子、小林剛、東谷篤志、石原直忠、武田伸一、曽我部正博、二川健
  • Organizer: 日本筋学会第3回学術集会
• [Presentation] 無重力による酸化ストレス産生を介した筋細胞内シグナル・トランスダクション (2017)
  • Author(s): 内田貴之、木森有希、二宮みゆき、坂下禎宏、安倍知紀、真板綾子、中尾玲子、平坂勝也、東谷篤志、小林剛、曽我部正博、武田伸一、二川健
  • Organizer: 第71回日本栄養・食糧学会大会
• [Presentation] Effects of dietary soy protein on unloading mediated muscle atrophy (2016)
  • Author(s): Takeshi Nikawa, Rie Hashimoto, Katsuya Hirasaka, Chiaki Yano, Kana Aibara, Takayuki Uchida, Tomoki Abe, Ayako Ohno(Maita), Kazuhito Akama
  • Organizer: 11th Asian Microgravity Symposium
  • Place of Presentation: 北海道大学（北海道・札幌市）
  • Year and Date: 2016-10-26
  • Int'l Joint Research
• [Presentation] 廃用性筋萎縮とユビキチンリガーゼ (2016)
  • Author(s): 二川健
  • Organizer: 第21回日本病態プロテアーゼ学会学術集会
  • Place of Presentation: 千里ライフサイエンスセンター（大阪府・豊中市）
  • Year and Date: 2016-08-05
  • Invited
• [Presentation] 筋萎縮における細胞内シグナルとその制御による筋萎縮治療 (2016)
  • Author(s): 二川健
  • Organizer: 第70回日本栄養・食糧学会大会
  • Place of Presentation: 武庫川女子大学（兵庫県・西宮市）
  • Year and Date: 2016-05-13
  • Invited
• [Presentation] Unloading環境による筋萎縮とその栄養学的治療法 (2016)
  • Author(s): 二川健
  • Organizer: 第23回アミノ酸セミナー：基礎と臨床を結ぶ会
  • Place of Presentation: 東京国際フォーラム（東京都・千代田区）
  • Invited
• [Book] 食品サイエンス (2017)
  • Author(s): 井田くるみ、内田貴之、赤間一仁、二川健
  • Publisher: 羊土社
• [Book] 「季刊 腎と骨代謝」特集 スペースサイエンス (2017)
  • Author(s): 二川健
  • Publisher: 日本メディカルセンター