遺伝子改変マウスを用いた抗癌剤耐性肺癌の新規治療法の開発

2015 Fiscal Year Research-status Report

• Project/Area Number: 15K09170
• Research Institution: Mie University
• Principal Investigator: Gabazza Esteban 三重大学, 医学(系)研究科(研究院), 教授 (00293770)
• Co-Investigator(Kenkyū-buntansha): 戸田 雅昭 三重大学, 医学(系)研究科(研究院), 講師 (10202201) ; GABAZZA CORINA 三重大学, 医学(系)研究科(研究院), 25:学術研究員 (10750656)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 肺癌 / 上皮細胞成長因子受容体 / チロシンキナーゼ阻害剤 / 耐性 / 肺癌マウスモデル

Outline of Annual Research Achievements

本研究では、EGFR-チロシンキナーゼ阻害薬に対する耐性を示す肺特異的ヒト「L858R」EGFR過剰発現する肺癌マウスモデルを用いて、EGFR細胞内シグナル伝達経路の構成因子のsiRNAが肺癌の耐性を克服できるか否かを検討する。「L858R」EGFR過剰発現による肺癌マウスモデルは既に開発済であったが、平成２７年度はEGFR-チロシンキナーゼ阻害薬に対する耐性を示す「L858R」EGFR過剰発現マウスモデルの作製に成功した。この耐性マウスを作製するため、[L858R＋新たに変異]EGFR RecBAC組換えBACクローンとCCSP組換えBACクローンを構築し、直鎖化し、精製した後C57BL/6J マウスから採取した受精卵にマイクロインジェクションを行った。

Current Status of Research Progress

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

Reason

平成２７年度はEGFR-チロシンキナーゼ阻害薬に対する耐性を示すEGFR過剰発現マウスモデルの作製に成功したので２８年度の研究目標は達成できる見込みである。

Strategy for Future Research Activity

１）伝達機構分子に対するsiRNAの濃度依存性の効果：A549細胞はEGFで刺激する前に伝達機構分子のsiRNA又は各々のscrambled siRNAの多数の濃度を培養液に添加し、24時間後に伝達機構分子の発現量をPCRで確認する。これらの蛋白の発現は他の方法（flow cytometry等）でも確認する。さらに、EGFR-TKI耐性の肺癌モデルマウス由来癌細胞はコラゲナーゼ法で分離し、同in vitro実験を行う。
２）EGFR-TKI耐性肺癌マウスに対する伝達機構分子のsiRNAによる耐性克服の効果の検討：EGFR-TKI耐性肺癌マウスに、週3回、伝達機構分子のsiRNA又各々タゲートのscrambled siRNAを経気管支的に投与し、2週間毎にマウス肺・全身のCT検査にて腫瘍の大きさなどを比較検討する。

Research Products

• [Int'l Joint Research] Stanford University/Department of Hematology(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Stanford University/Department of Hematology
• [Int'l Joint Research] Illinois University, at Champaine/Department of Microbiology(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Illinois University, at Champaine/Department of Microbiology
• [Journal Article] Improvement in early diagnosis of Japanese spotted fever by using a novel Rick PCR system. (2015)
  • Author(s): Kondo M, Akachi S, Kawano M, Yamanaka K, Yamagiwa A, Gabazza EC, Ando K, Mizutani H.
  • Journal Title: J Dermatol Volume: 42 Pages: 1066-1071
  • DOI: 10.1111/1346-8138.13015.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Usefulness of Small Intestinal Endoscopy in a Case of Adult-onset Familial Mediterranean Fever Associated with Jejunoileitis. (2015)
  • Author(s): Kitade T, Horiki N, Katsurahara M, Totoki T, Harada T, Tano S, Yamada R, Hamada Y, Inoue H, Tanaka K, Gabazza EC, Hayashi H, Tanaka M, Takei Y.
  • Journal Title: Intern Med Volume: 54 Pages: 1343-1347
  • DOI: 10.2169/internalmedicine.54.3690.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Activated protein C modulates the proinflammatory activity of dendritic cells. (2015)
  • Author(s): Matsumoto T, Matsushima Y, Toda M, Roeen Z, D'Alessandro-Gabazza CN, Hinneh JA, Harada E, Yasuma T, Yano Y, Urawa M, Kobayashi T, Taguchi O, Gabazza EC.
  • Journal Title: J Asthma Allergy Volume: 8 Pages: 29-37
  • DOI: 10.2147/JAA.S75261.
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Effects of sugar-sweetened beverage intake on the development of type 2 diabetes mellitus in subjects with impaired glucose tolerance: the Mihama diabetes prevention study. (2015)
  • Author(s): Teshima N, Shimo M, Miyazawa K, Konegawa S, Matsumoto A, Onishi Y, Sasaki R, Suzuki T, Yano Y, Matsumoto K, Yamada T, Gabazza EC, Takei Y, Sumida Y.
  • Journal Title: J Nutr Sci Vitaminol (Tokyo) Volume: 61 Pages: 14-19
  • DOI: 10.3177/jnsv.61.14.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Amelioration of Atherosclerosis by the New Medicinal Mushroom Grifola gargal Singer. (2015)
  • Author(s): Harada E, D'Alessandro-Gabazza CN, Toda M, Morizono T, Chelakkot-Govindalayathil AL, Roeen Z, Urawa M, Yasuma T, Yano Y, Sumiya T, Gabazza EC
  • Journal Title: J Med Food Volume: 18 Pages: 872-881
  • DOI: 10.1089/jmf.2014.3315.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Endoscopic findings and lesion distribution in amebic colitis. (2015)
  • Author(s): Horiki N, Furukawa K, Kitade T, Sakuno T, Katsurahara M, Harada T, Tano S, Yamada R, Hamada Y, Inoue H, Tanaka K, Gabazza EC, Ishii N, Fukuda K, Omata F, Fujita Y, Tachibana H, Takei Y.
  • Journal Title: J Infect Chemother Volume: 21 Pages: 444-448
  • DOI: 10.1016/j.jiac.2015.02.004.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Second and third-look endoscopy for the prevention of post-ESD bleeding. (2015)
  • Author(s): Tano S, Horiki N, Omata F, Tanaka K, Hamada Y, Katsurahara M, Ninomiya K, Nishikawa K, Nojiri K, Yamada R, Inoue H, Gabazza EC, Katayama N, Takei Y.
  • Journal Title: Medicine (Baltimore) Volume: 94 Pages: e491
  • DOI: 10.1097/MD.0000000000000491.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Olopatadine, a non-sedating H1 antihistamine, decreases the nocturnal scratching without affecting sleep quality in atopic dermatitis. (2015)
  • Author(s): Yamanaka K, Motomura E, Noro Y, Umeda K, Morikawa T, Umeda-Togami K, Omoto Y, Isoda K, Kondo M, Tsuda K, Okuda M, Gabazza EC, Mizutani H.
  • Journal Title: Exp Dermatol Volume: 24 Pages: 227-229
  • DOI: 10.1111/exd.12630.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Thrombomodulin inhibits the activation of eosinophils and mast cells. (2015)
  • Author(s): Roeen Z, Toda M, D'Alessandro-Gabazza CN, Onishi M, Kobayashi T, Yasuma T, Urawa M, Taguchi O, Gabazza EC
  • Journal Title: Cell Immunol Volume: 293 Pages: 34-40
  • DOI: 10.1016/j.cellimm.2014.11.005.
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Is there still hope for single therapies: how do we set up experimental systems to efficiently test combination therapies? (2015)
  • Author(s): Joos GF, Gabazza EC
  • Journal Title: Respirology Volume: 20 Pages: 15-23
  • DOI: 10.1111/resp.12438.
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Protein S exacerbates alcoholic hepatitis by stimulating liver natural killer T cells. (2015)
  • Author(s): Chelakkot-Govindalayathil AL, Mifuji-Moroka R, D'Alessandro-Gabazza CN, Toda M, Matsuda Y, Gil-Bernabe P, Roeen Z, Yasuma T, Yano Y, Gabazza EC, Iwasa M, Takei Y
  • Journal Title: J Thromb Haemost Volume: 13 Pages: 142-154
  • DOI: 10.1111/jth.12789.
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Targeting Periostin for the Treatment of Pulmunary Fibrosis (2015)
  • Author(s): Gabazza EC, Kobayashi T, D’Alessandro-Gabazza CN, Toda M, Roeen Z, Takahashi Y, Fujiwara K, Hinneh JA, Yasuma T, Taguchi O
  • Organizer: 9th International Symposium on Nanomedicine (ISNM2015)
  • Place of Presentation: 三重大学
  • Year and Date: 2015-12-10 – 2015-12-12
  • Int'l Joint Research
• [Presentation] Matrix metalloproteinase-2 is protective in bleomycin-induced pulmonary fibrosis (2015)
  • Author(s): Tomaru A, Gabazza E, Kobayashi T, Kobayashi H, Taguchi O, Takagi T, Oonishi M, Fujiwara K, D'Alessandro Gabazza C, Takahashi Y, Urata K
  • Organizer: ERS AMSTERDAM, Netherlands
  • Place of Presentation: オランダ アムステルダム
  • Year and Date: 2015-09-26 – 2015-09-30
  • Int'l Joint Research
• [Presentation] Lung overexpression of matrix metalloproteinase-2 protects against bronchial asthma (2015)
  • Author(s): Fujiwara K, Urata K, Tomaru A, Takahashi Y, Onishi M, Takehiro Takagi, Kobayashi H, D'Alessandro Gabazza CN, Gabazza EC, Taguchi O, Kobayashi T
  • Organizer: ERS AMSTERDAM, Netherlands
  • Place of Presentation: オランダ アムステルダム
  • Year and Date: 2015-09-26 – 2015-09-30
  • Int'l Joint Research
• [Presentation] The immunomodulatory activity of protein S protects against allergic airway inflammation. (2015)
  • Author(s): Takahashi Y, Urata K, Tomaru A, Fujiwara K, Onishi M, Takagi T, Kobayashi H, D'Alessandro-Gabazza C, Gabazza E, Osamu Taguchi O, Kobayashi T
  • Organizer: ERS AMSTERDAM, Netherlands
  • Place of Presentation: オランダ アムステルダム
  • Year and Date: 2015-09-26 – 2015-09-30
  • Int'l Joint Research
• [Presentation] Clinical significance of transcutaneous PCO2 in patients with obstructive sleep apnea syndrome. (2015)
  • Author(s): Sakaguchi T, Hataji O, Saiki H, Gabazza EC, D'Alessandro-Gabazza C, Nishii Y, Kobayashi T, Taguchi O
  • Organizer: ERS AMSTERDAM, Netherlands
  • Place of Presentation: オランダ アムステルダム
  • Year and Date: 2015-09-26 – 2015-09-30
  • Int'l Joint Research
• [Presentation] Clinical severity of chronic obstructive pulmonary disease is associated with bone mineral density. (2015)
  • Author(s): Saiki H, Hataji O, Sakaguchi T, Gabazza EC, Nishii Y, D'Alessandro-Gabazza C, Kobayashi T, Taguchi O
  • Organizer: ERS AMSTERDAM, Netherlands
  • Place of Presentation: オランダ アムステルダム
  • Year and Date: 2015-09-26 – 2015-09-30
  • Int'l Joint Research
• [Presentation] Tracheobronchial reconstruction with implantation of the right main bonchus: Surgical technique in the treatment of tracheal adenoid cystic carcinoma. (2015)
  • Author(s): Watanabe F, Takeo M, Tenpaku H, Yada I, Itou K, Naitou M, Hataji O, D’Alessandro-Gabazza CN, Gabazza EC, Shinpo H
  • Organizer: ESTS Abstract Submission
  • Place of Presentation: ポルトガル リスボン
  • Year and Date: 2015-05-31 – 2015-06-03
  • Int'l Joint Research
• [Book] Annual Review 2016 呼吸器 (2016)
  • Author(s): 小林 哲、ガバザエステバン
  • Total Pages: 233(p6-11)
  • Publisher: 中外医薬社
• [Patent(Industrial Property Rights)] 肺特異的にヒトＥＧＦＲを発現する非ヒト哺乳類のトランスジェニック動物 (2015)
  • Inventor(s): ガバザ エステバン、ガバザ コリナ
  • Industrial Property Rights Holder: ガバザ エステバン、ガバザ コリナ
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 特願2015-158369
  • Filing Date: 2015-08-10