低酸素ストレスに対するシステミックな生体応答機構の解明

2014 Fiscal Year Annual Research Report

• Project Area: Oxygen biology: a new criterion for integrated understanding of life
• Project/Area Number: 26111002
• Research Institution: Tohoku University
• Principal Investigator: 山本 雅之 東北大学, 医学(系)研究科(研究院), 教授 (50166823)
• Co-Investigator(Kenkyū-buntansha): 鈴木 教郎 東北大学, 医学(系)研究科(研究院), 講師 (20447254)
• Project Period (FY): 2014-06-27 – 2019-03-31
• Keywords: 造血 / エリスロポエチン / 転写制御

Outline of Annual Research Achievements

酸素は生体がエネルギーを得るために必須であり、組織への酸素供給量の低下は、生命の存続を脅かす危険な「低酸素ストレス」状態を惹起する。低酸素ストレスは、貧血による酸素供給不足や腫瘍内の血管低形成などによって発生し、病態と深く関連する。本研究では、低酸素ストレスに対する生体防御系である赤血球増産による酸素供給能亢進の分子メカニズム解明を試みている。
初年度は、赤血球増殖因子エリスロポエチンの産生機構について、遺伝子改変マウスを用いた解析を行った。その結果、低酸素誘導性転写因子のひとつであるHIF2aが肝臓および腎臓における低酸素誘導性のエリスロポエチン産生において必須であることが明らかとなった。また、HIF2aの抑制因子であるPHDが肝臓と腎臓では異なるメカニズムでエリスロポエチン遺伝子の転写を制御していることを明らかにした。さらに、エリスロポエチン産生能が著しく低下した慢性貧血マウスを用いて、造血における鉄代謝制御系について解析を進めた。本マウスでは、造血活性が低下しており、鉄利用を抑制するホルモンであるヘプシジンが肝臓から大量に分泌されており、貯蔵鉄が蓄積していた。この鉄蓄積とヘプシジン産生亢進は、エリスロポエチン製剤を本マウスに投与することにより、速やかに改善された。以上の結果から、低酸素によるエリスロポエチン産生誘導の分子機序が明らかとなり、造血制御と鉄代謝系が密接に関係していることが理解された。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

エリスロポエチン遺伝子の発現制御機構について、遺伝子改変マウスの作出が順調に進み、一定の成果を得た。また、腎臓におけるPHD-HIF制御系の役割に関する成果について論文投稿を行い、受理された。肝臓での制御機構についても投稿先から好意的なコメントを得ている。

Strategy for Future Research Activity

慢性貧血マウスを用いて、貧血による低酸素の影響について個体レベルでの解析を進める。また、造血を活性化した際の鉄動態や再酸素化の影響についてもマウスモデルを用いた解析を行う。エリスロポエチン遺伝子の制御機構については、腎臓特異的な制御機構の研究を進める。

Research Products

• [Journal Article] Identification of a functional antioxidant response element within the eighth intron of the human ABCC3 gene. (2015)
  • Author(s): Canet MJ, Merrell MD, Harder BG, Maher JM, Wu T, Lickteig AJ, Jackson JP, Zhang DD, Yamamoto M, and Cherrington NJ.
  • Journal Title: Drug Metab Dispos Volume: 43 Pages: 93-99
  • DOI: 10.1124/dmd.114.060103
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] MAFG is a transcriptional repressor of bile acid synthesis and metabolism. (2015)
  • Author(s): de Aguiar Vallim TQ, Tarling EJ, Ahn H, Hagey LR, Romanoski CE, Lee RG, Graham MJ, Motohashi H, Yamamoto M, and Edwards PA.
  • Journal Title: Cell Metab Volume: 21 Pages: 298-310
  • DOI: 10.1016/j.cmet.2015.01.007
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The Keap1-Nrf2 system and diabetes mellitus. (2015)
  • Author(s): Uruno A, Yagishita Y, and Yamamoto
  • Journal Title: Arch Biochem Biophys Volume: 566C Pages: 76-84
  • DOI: 10.1016/j.abb.2014.12.012
  • Peer Reviewed
• [Journal Article] In vivo regulation of erythropoiesis by chemically inducible dimerization of the erythropoietin receptor intracellular domain. (2015)
  • Author(s): Suzuki N, Mukai HY, Yamamoto M.
  • Journal Title: PLoS One Volume: 10 Pages: e0119442
  • DOI: 10.1371/journal.pone.0119442
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] DNA methyltransferase 3a regulates osteoclast differentiation by coupling to an S-adenosylmethionine-producing metabolic pathway. (2015)
  • Author(s): Nishikawa K, Iwamoto Y, Kobayashi Y, Katsuoka F, Kawaguchi S, Tsujita T, Nakamura T, Kato S, Yamamoto M, Takayanagi H, Ishii M.
  • Journal Title: Nat. Med. Volume: 21 Pages: 281-287
  • DOI: 10.1038/nm.3774
  • Peer Reviewed
• [Journal Article] CNC-bZIP Protein Nrf1-Dependent Regulation of Glucose-Stimulated Insulin Secretion. (2015)
  • Author(s): Zheng H, Fu J, Xue P, Zhao R, Dong J, Liu D, Yamamoto M, Tong Q, Teng W, Qu W, Zhang Q, Andersen ME, Pi J.
  • Journal Title: Antioxid. Redox Signal. Volume: 22 Pages: 819-831
  • DOI: 10.1089/ars.2014.6017.
• [Journal Article] Hypoxia-sensitive reporter system for high-throughput screening. (2015)
  • Author(s): Tsujita T, Kawaguchi S, Dan T, Baird L, Miyata T, Yamamoto M.
  • Journal Title: Tohoku J. Exp. Med. Volume: 235 Pages: 151-159
  • DOI: 10.1620/tjem.235.151.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Alcohol dehydrogenase 3 contributes to the protection of liver from nonalcoholic steatohepatitis. (2015)
  • Author(s): Goto M, Kitamura H, Alam MM, Ota N, Haseba T, Akimoto T, Shimizu A, Takano-Yamamoto T, Yamamoto M, Motohashi H.
  • Journal Title: Genes Cells Volume: - Pages: -
  • DOI: 10.1111/gtc.12237
  • Peer Reviewed
• [Journal Article] The Human GATA1 Gene Retains a 5' Insulator that Maintains Chromosomal Architecture and GATA1 Expression Levels in Splenic Erythroblasts. (2015)
  • Author(s): Moriguchi T, Yu L, Takai J, Hayashi M, Satoh H, Suzuki M, Ohneda K, Yamamoto M.
  • Journal Title: Mol. Cell. Biol. Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] Progenitor stage-specific activity of a cis-acting double GATA motif for Gata1 gene expression. (2015)
  • Author(s): Moriguchi T, Suzuki M, Yu L, Takai J, Ohneda K, Yamamoto M.
  • Journal Title: Mol. Cell. Biol. Volume: 35 Pages: 805-815
  • DOI: 10.1128/MCB.01011-14.
  • Peer Reviewed
• [Journal Article] Erythropoietin gene expression: developmental-stage specificity, cell-type specificity, and hypoxia inducibility. (2015)
  • Author(s): Suzuki N.
  • Journal Title: Tohoku J. Exp. Med. Volume: 235 Pages: 233-240
  • DOI: 10.1620/tjem.235.233
  • Peer Reviewed
• [Journal Article] Erythropoietin Synthesis in Renal Myofibroblasts Is Restored by Activation of Hypoxia Signaling. (2015)
  • Author(s): Souma T, Nezu M, Nakano D, Yamazaki S, Hirano I, Sekine H, Dan T, Takeda K, Fong GH, Nishiyama A, Ito S, Miyata T, Yamamoto M, Suzuki N
  • Journal Title: J. Am. Soc. Nephrol. Volume: - Pages: -
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Transcription factor Nrf1 negatively regulates the cystine/glutamate transporter and lipid-metabolizing enzymes. (2014)
  • Author(s): Tsujita T, Mullin V, Baird L, Matsuyama Y, Takaku M, Walsh SV, Griffin JL, Uruno A, Yamamoto M, Hayes JD
  • Journal Title: Mol. Cell. Biol. Volume: 34 Pages: 3800-3816
  • DOI: 10.1128/MCB.00110-14.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Validation of multiple single nucleotide variation calls by additional exome analysis with a semiconductor sequencer to supplement data of whole-genome sequencing of a human population. (2014)
  • Author(s): Motoike IN, Matsumoto M, Danjoh I, Katsuoka F, Kojima K, Nariai N, Sato Y, Yamaguchi-Kabata Y, Ito S, Kudo H, Nishijima I, Nishikawa S, Pan X, Saito R, Saito S, Saito T, Shirota M, Tsuda K, Yokozawa J, Igarashi K, Minegishi N, Tanabe O, Fuse N, Nagasaki M, Kinoshita K, Yasuda J, Yamamoto M.
  • Journal Title: BMC Genomics Volume: 15 Pages: 673
  • DOI: 10.1186/1471-2164-15-673.
  • Peer Reviewed / Open Access
• [Journal Article] Remarkable induction of UV-signature mutations at the 3'-cytosine of dipyrimidine sites except at 5'-TCG-3' in the UVB-exposed skin epidermis of xeroderma pigmentosum variant model mice (2014)
  • Author(s): Ikehata H, Chang Y, Yokoi M, Yamamoto M, Hanaoka F.
  • Journal Title: DNA Repair Volume: 22 Pages: 112-122
  • DOI: 10.1016/j.dnarep.2014.07.012.
  • Peer Reviewed
• [Journal Article] GATA2 regulates differentiation of bone marrow-derived mesenchymal stem cells. (2014)
  • Author(s): Kamata M, Okitsu Y, Fujiwara T, Kanehira M, Nakajima S, Takahashi T, Inoue A, Fukuhara N, Onishi Y, Ishizawa K, Shimizu R, and Yamamoto M, Harigae H.
  • Journal Title: Haematologica Volume: 99 Pages: 1686-1696
  • DOI: 10.3324/haematol.2014.105692
  • Peer Reviewed
• [Journal Article] An efficient quantitation method of next-generation sequencing libraries by using MiSeq sequencer. (2014)
  • Author(s): Katsuoka F, Yokozawa J, Tsuda K, Ito S, Pan X, Nagasaki M, Yasuda J, Yamamoto M.
  • Journal Title: Anal. Biochem. Volume: 466 Pages: 27-29
  • DOI: 10.1016/j.ab.2014.08.015.
  • Peer Reviewed
• [Journal Article] Nrf2 induces fibroblast growth factor 21 in diabetic mice. (2014)
  • Author(s): Furusawa Y, Uruno A, Yagishita Y, Higashi C, Yamamoto M.
  • Journal Title: Genes Cells Volume: 19 Pages: 864-878
  • DOI: 10.1111/gtc.12186.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Activation of the NRF2 pathway and its impact on the prognosis of anaplastic glioma patients. (2014)
  • Author(s): Kanamori M, Higa T, Sonoda Y, Murakami S, Dodo M, Kitamura H, Taguchi K, Shibata T, Watanabe M, Suzuki H, Shibahara I, Saito R, Yamashita Y, Kumabe T, Yamamoto M, Motohashi H, Tominaga T.
  • Journal Title: Neuro. Oncol. Volume: 17 Pages: 555-565
  • DOI: 10.1093/neuonc/nou282
  • Peer Reviewed
• [Journal Article] Nuclear factor erythroid 2-related factor 2 regulates toll-like receptor 4 innate responses in mouse liver ischemia-reperfusion injury through Akt-forkhead box protein O1 signaling network. (2014)
  • Author(s): Huang J, Yue S, Ke B, Zhu J, Shen XD, Zhai Y, Yamamoto M, Busuttil RW, Kupiec-Weglinski JW.
  • Journal Title: Transplantation Volume: 98 Pages: 721-728
  • DOI: 10.1097/TP.0000000000000316.
  • Peer Reviewed
• [Presentation] エリスロポエチン産生細胞の正常と病態 (2015)
  • Author(s): 鈴木教郎
  • Organizer: 第２回 日本腎臓研究会
  • Place of Presentation: 経団連会館（東京）
  • Year and Date: 2015-01-17 – 2015-01-17
  • Invited
• [Presentation] 環境ストレスへの生体応答の分子基盤 (2014)
  • Author(s): 山本雅之
  • Organizer: 第56回日本先天代謝異常学会総会
  • Place of Presentation: 江陽グランドホテル（仙台市）
  • Year and Date: 2014-11-13 – 2014-11-13
  • Invited
• [Presentation] Genetic activation of hypoxia-signaling restores erythropoietin synthesis in renal myofibroblasts. (2014)
  • Author(s): 5.Souma T, Nezu M, Ito S, Suzuki N, and Yamamoto M.
  • Organizer: ASN renal week
  • Place of Presentation: Philadelphia, Pennsylvania, USA
  • Year and Date: 2014-11-11 – 2014-11-16
• [Presentation] エリスロポエチン産生細胞とエリスロポエチン遺伝子の発現制御機構 (2014)
  • Author(s): 山本雅之
  • Organizer: 日本血液学会学術集会
  • Place of Presentation: 大阪国際会議場（大阪市）
  • Year and Date: 2014-11-02 – 2014-11-02
  • Invited
• [Presentation] 酸素と医学 (2014)
  • Author(s): 山本雅之
  • Organizer: レドックスシンポジウム 酸素生物学の誕生
  • Place of Presentation: 名古屋大学（名古屋市）
  • Year and Date: 2014-10-11 – 2014-10-11
  • Invited
• [Presentation] Molecular Basis of the Keap1-Nrf2 System Regulating Environmental Stress Response (2014)
  • Author(s): Masayuki Yamamoto
  • Organizer: Nobel Conference
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2014-10-08 – 2014-10-08
  • Invited
• [Presentation] エリスロポエチン産生細胞と腎疾患 (2014)
  • Author(s): 山本雅之
  • Organizer: 第45回新潟腎カンファランス
  • Place of Presentation: 新潟大学（新潟市）
  • Year and Date: 2014-09-25 – 2014-09-25
  • Invited
• [Presentation] Erythropoietin production in neural and neural crest cells during primitive erythropoiesis (2014)
  • Author(s): Masayuki Yamamoto
  • Organizer: 19th Hemoglobin Switching Conference
  • Place of Presentation: Oxford, UK
  • Year and Date: 2014-09-05 – 2014-09-05
  • Invited