Identification of driver genes in lung cancer through a semi-genome wide shRNA library screen based on anoikis resistance phenotype

• Project/Area Number: 26293197
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Respiratory organ internal medicine
• Research Institution: Nagoya University
• Principal Investigator: Sato Mitsuo 名古屋大学, 医学部附属病院, 講師 (70467281)
• Co-Investigator(Kenkyū-buntansha): 近藤 征史 名古屋大学, 医学(系)研究科(研究院), 准教授 (00378077) ; 長谷 哲成 名古屋大学, 医学部附属病院, 助教 (30621635)
• Co-Investigator(Renkei-kenkyūsha): YUKAWA Hiroshi 名古屋大学, 大学院工学系研究科, 特任講師 (30634646) ; YOKOI Kohei 名古屋大学, 大学院医学系研究科, 教授 (60378007)
• Research Collaborator: YOGO Naoyuki 名古屋大学, 大学院工学研究科, 研究員 (70817874)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000); Fiscal Year 2016: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000); Fiscal Year 2014: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
• Keywords: 肺癌 / アノイキス / プールshRNAライブラリー / プロテアソーム / 正常ヒト気管支上皮細胞発癌モデル（HBEC） / 国際研究者交流 / プールshRNA / 不死化正常ヒト気管支上皮細胞 / 変異KRAS / p53

Outline of Final Research Achievements

This study was aimed to identify novel therapeutic targets for lung cancer. To this end, we employed two approached. First, we tried to identify genes that contribute to anoikis resistance by doing shRNA library screen using an immortalized normal human bronchial epithelial cell line, HBEC. Anoikis is a special type of apoptosis induced by loss of attachment to extra cellular matrix, and resistance against anoikis is one of critical malignant properties of cancer cells. We identified RNF146 as a gene whose loss could contribute to anoikis resistance in HBEC. However, this result was not reproduced in lung cancer cell lines. Second, we did an integrated analysis combining a shRNA library screen with utilization of gene expression and copy number analysis using an aggressive lung cancer cell line, H460. We identified proteasome subunit gene PSMA6 as a novel therapeutic target for lung cancer.

Academic Significance and Societal Importance of the Research Achievements

肺癌は難治癌の代表であり、新しい有効な治療方法の開発は社会的に重要な課題である。本研究は肺癌の新規治療標的の発見を目的とした。肺癌細胞株を用いて、プールshRNAライブラリーによる網羅的ノックダウン手法を用いた機能的なスクリーニングと肺癌における遺伝子発現量およびコピー数の統合解析を実施した。その結果、肺癌の新規治療標的としてプロテアソームサブユニット遺伝子PSMA6を同定し、結果を論文発表した。

Research Products

• [Int'l Joint Research] Univ. of Texas Southwestern Medical Ctr./MD Anderson Cancer Center/Ohio State University, Columbus(米国)
• [Journal Article] Identification of Proteasomal Catalytic Subunit PSMA6 as a Therapeutic Target for Lung Cancer. (2017)
  • Author(s): Kakumu T, Sato M, Goto D, Kato T, Yogo N, Hase T, Morise M, Fukui T, Yokoi K, Sekido Y, Girard L, Minna JD, Byers LA, Heymach JV, Coombes KR, Kondo M, Hasegawa Y.
  • Journal Title: Cancer Sci. Volume: 108 Issue: 4 Pages: 732-743
  • DOI: 10.1111/cas.13185
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] ZEB1 drives epithelial-to-mesenchymal transition in lung cancer. (2016)
  • Author(s): Larsen JE, Nathan V, Osborne JK, Farrow RK, Deb D, Sullivan JP, Dospoy PD, Augustyn A, Hight SK, Sato M, Girard L, Behrens C, Wistuba II, Gazdar AF, Hayward NK, Minna JD.
  • Journal Title: J Clin Invest. Volume: 126 Issue: 9 Pages: 3219-35
  • DOI: 10.1172/jci76725
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Growth inhibitory effects of miR-221 and miR-222 in non-small cell lung cancer cells. (2015)
  • Author(s): Yamashita R, Sato M, Kakumu T, Hase, T, Yogo, N, Maruyama E, Sekido Y, Kondo M, Hasegawa, Y.
  • Journal Title: Cancer Medicine Volume: 4 Issue: 4 Pages: 551-564
  • DOI: 10.1002/cam4.412
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nuclear Receptor Expression and Function in Human Lung Cancer Pathogenesis. (2015)
  • Author(s): Kim J, Sato M, Choi JW, Kim HW, Yeh BI, Larsen JE, Minna JD, Cha JH, Jeong Y.
  • Journal Title: PLoS One Volume: 10 Issue: 8 Pages: e0134842-e0134842
  • DOI: 10.1371/journal.pone.0134842
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Identification of Proteasomal Catalytic Subunit PSMA6 as a Therapeutic Target for Lung Cancer (2016)
  • Author(s): 佐藤光夫、各務智彦、加藤俊夫、與語直之、長谷哲成、森 瀬昌宏、福井高幸、横井香平、Luc Girard 、John Minna、 近藤征史、長谷川好規
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市）
• [Presentation] Cytochrome c oxidase subunit 5a (COX5A) is identified as a potential therapeutic target for lung cancer with high therapeutic index through a pooled shRNA screen (2016)
  • Author(s): 1.Toshio Kato, Mitsuo Sato, Masashi Kondo, Tomohiko Kakumu, Naoyuki Yogo, Tetsunari Hase, Masahiro Morise, John D Minna, Yoshinori Hasegawa
  • Organizer: アメリカ癌学会年次総会
  • Place of Presentation: ニューオリンズ（アメリカ合衆国）
  • Int'l Joint Research
• [Presentation] 肺がん治療標的としてのCDK11 の可能性 (2015)
  • Author(s): 各務智彦、佐藤光夫、加藤俊夫、與語直之、長谷哲成、森瀬昌宏、近藤政史、関戸好孝、長谷川好規
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 国際会議場（愛知県名古屋市９
  • Year and Date: 2015-10-08
• [Presentation] Tumor suppressive roles of miR-221 and miR-222 in lung cancer (2014)
  • Author(s): Yamashita R, Sato M, Kakumu T, Hase T, Yogo N, Maruyama E, Sekido Y, Kondo M, Hasegawa Y.
  • Organizer: ENA-2014 Symposium
  • Place of Presentation: バルセロナ、スペイン
  • Year and Date: 2014-11-19
• [Presentation] 非小細胞肺癌治療薬としてのmiR-221とmiR-222の可能性 (2014)
  • Author(s): 佐藤光夫、長谷哲成、近藤征史、長谷川好規
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 神奈川県、横浜市、パシフィコ横浜
  • Year and Date: 2014-09-25