Potential of new anti-cancer agents targeting the nuclear translocation signaling of HB-EGF C-terminal fragments

• Project/Area Number: 22590704
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Nagoya City University
• Principal Investigator: TANIDA Satoshi 名古屋市立大学, 大学院・医学研究科, 助教 (30528782)
• Co-Investigator(Kenkyū-buntansha): JOH Takashi 名古屋市立大学, 大学院・医学研究科, 教授 (30231369) ; KATAOKA Hiromi 名古屋市立大学, 大学院・医学研究科, 准教授 (40381785)
• Co-Investigator(Renkei-kenkyūsha): HIGASHIYAMA Shigeki 愛媛大学, 大学院・医学系研究科, 教授 (60202272)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2012: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: EGFファミリー / HB-EGF核移行シグナル / PLZF / Alphascreen assay / IL-8 / HB-EGF-CTF / colitic cancer / 潰瘍性大腸炎 / 炎症性腸疾患

Research Abstract

Current treatment target toward advanced colorectal cancers is mainly focused on the epidermal growth factor receptor (EGFR) signaling, but its additive effects with chemotherapy are still limited. A disintegrin and metalloproteinase (ADAM) cleaves the proheparin-binding epidermal growth factor like growth factor (proHB-EGF). And soluble HB-EGF activates EGFR. In parallel, the carboxy-terminal fragment of proHB-EGF (HB-EGF-CTF) translocate into the inner nuclear membrane, subsequently exerts on the regulation of cell proliferation by binding nuclear promyelocytic leukemia zinc finger (PLZF) protein, a transcriptional repressor, thereby causing its nuclear export. We hypothesized that the inhibition of HB-EGF-CTF nuclear translocation may be a new strategy in preventing cell proliferation.Methods:12-O-tetradecanoylphorbor-13-acetate (TPA) was treated to activate ADAM. Nine-thousand chemical compounds were screened for their efficacies in blocking the binding of HB-EGF-CTF to promyelocyt ic leukemia zinc finger (PLZF) with Alphascreen system. The obtained candidates were then used to block the binding of HB-EGF-CTF to PLZF in colon cancer cells, HT29 and HCT116. Cell proliferation was investigated with a growth curve assay. The intracellular localization, and association between HB-EGF-CTF and PLZF, was assessed with immunofluorescent staining, and immunoprecipitation and Western blotting, respectively. The effects of obtained candidates on EGFR phosphorylation and on nuclear translocation of HB-EGF-CTF and export of PLZF during the angiotensin II type1 receptor (AT1R) knockdown were also investigated. Results: Telmisartan and candesartan were found to be potential candidates. Telmisartan inhibited TPA-induced cell proliferation stronger than candesartan. Telmisartan, but not candesartan blocked the nuclear translocation of HB-EGF-CTF, and binding of HB-EGF-CTF to PLZF, during TPA stimulation. Both telmisartan and candesartan did not inhibit TPA-induced EGFR phosphorylation, and telmisartan, but not candesartan, inhibited TPA-induced nuclear translocation of HB-EGF-CTF after knockdown of AT1R. Conclusions: The inhibition of HB-EGF-CTF nuclear translocation with telmisartan may be a novel strategy in preventing cell proliferation.

Research Products

• [Journal Article] Telmisartan inhibits cell proliferation by blocking nuclear translocation of ProHB-EGF C-terminal fragment in colon cancer cells. (2013)
  • Author(s): Ozeki K, Tanida S, Morimoto C, Inoue Y, Mizoshita T, Tsukamoto H, Shimura T, Kataoka H, Kamiya T, Nishiwaki E, Ishiguro H, Higashiyama S, Joh T.
  • Journal Title: PLoS One Volume: 8(2) Issue: 2 Pages: e56770-e56770
  • DOI: 10.1371/journal.pone.0056770
  • URL: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3579940/
  • Peer Reviewed
• [Journal Article] Involvement of cell proliferation induced by dual intracellular signaling of HB-EGF in the development of colitis-associated cancer duringulcerative colitis (2011)
  • Author(s): Tanida S, Mizoshita T, Mizushima T, Shimura T, Kamiya T, Kataoka H, Joh T
  • Journal Title: Ulcers Volume: volume 2011 Pages: 6-6
  • DOI: 10.1155/2011/457637
  • Peer Reviewed
• [Journal Article] IBD 腸管炎症に関わる炎症性サイトカインによるEGF シグナルを介した大腸細胞増殖機序 (2011)
  • Author(s): 谷田諭史、尾関啓司,塚本宏延 、溝 下 勤、片岡洋望、神 谷 武、城 卓志
  • Journal Title: 潰瘍 Volume: 38(2) Pages: 135-138
• [Journal Article] HB-EGF-C末端核移行シグナルをターゲットにした新規薬剤探索と細胞増殖抑制機序 (2011)
  • Author(s): 尾関啓司,谷田諭史,溝下 勤,水島隆史,志村貴也,村上賢治,平田慶和,片岡洋望,神谷 武,福田伸治,東山繁樹,城 卓志
  • Journal Title: 消化器と免疫 Volume: 47 Pages: 144-146
• [Journal Article] TGFβinduces proHB-EGF shedding and EGFR transactivation through ADAM activation in gastric cancercells (2010)
  • Author(s): Ebi M, Kataoka H, Shimura T, Kubota E, Hirata Y, Mizushima T, Mizoshita T, Tanaka M, Mabuchi M, Tsukamoto H, Tanida S, Kamiya T, Higashiyama S, Joh T
  • Journal Title: Biochem Biophys Res Commun Volume: 402(3) Issue: 3 Pages: 449-54
  • DOI: 10.1016/j.bbrc.2010.09.130
  • Peer Reviewed
• [Journal Article] Intranuclear translocation signaling of HB-EGF carboxy-terminal fragment and mucosal defense through cell proliferation and migration in digestive tracts (2010)
  • Author(s): Tanida S, Kataoka H, Mizoshita T, Shimura T, Kamiya T, Joh T
  • Journal Title: Digestion Volume: 82(3) Issue: 3 Pages: 145-9
  • DOI: 10.1159/000310903
  • Peer Reviewed
• [Journal Article] Intranuclear translocation signaling of HB-EGF C terminal fragment and mucosal defense through cell proliferation and migration in digestive tracts (2010)
  • Author(s): Satoshi Tanida
  • Journal Title: Digestion Volume: 82 Pages: 145-149
  • Peer Reviewed
• [Journal Article] BCL6 degradation caused by the interation with the C-terminus of proHB-EGF induces cycline D2 expression in gastric cancers (2009)
  • Author(s): Hirata Y, Ogasawara N, Sasaki M, Mizushima T, Mizoshita T, Shimura T, Mori Y, Kubota E, Wada T, Tanida S, Kataoka H, Kamiya T, Higashiyama S, Joh T
  • Journal Title: British Journal of Cancer Volume: 100 Issue: 5 Pages: 1320-1329
  • DOI: 10.1016/s0016-5085(09)61447-5
  • Peer Reviewed
• [Presentation] 大腸癌増殖シグナルHB-EGF-C 末端核移行をターゲットとした新規薬剤探索 (2012)
  • Author(s): 尾関啓司、谷田諭史、溝 下 勤、塚本宏延、片岡洋望、城 卓志.
  • Organizer: 第50回日本癌治療学会学術集会
  • Place of Presentation: 横浜
  • Year and Date: 2012-10-25
• [Presentation] 炎症性サイトカイン(IL-8)によるHB-EGF-C末端核移行シグナルを介した大腸細胞増殖機序 (2012)
  • Author(s): 尾関啓司、谷田諭史、溝下 勤、塚本宏延、片岡洋望、城 卓志
  • Organizer: 第40回日本潰瘍学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-14
• [Presentation] TGFβによるROSおよびADAM17を介したEGF受容体transactivationのメカニズムの解析 (2011)
  • Author(s): 海老正秀、片岡洋望、志村貴也、溝下 勤、谷田諭史、平田慶和、神谷 武、水島隆史、東山繁樹、城 卓志
  • Organizer: JDDW 2011
  • Place of Presentation: 福岡
  • Year and Date: 2011-10-21
• [Presentation] IBD 腸管炎症に関わる炎症性サイトカインによるEGF-C末端signalを介した大腸癌細胞増殖機序 HB-EGF-CTFシグナル抑制薬網羅的探索から- (2011)
  • Author(s): 尾関啓司、谷田諭史、溝下 勤、塚本宏延、城 卓志
  • Organizer: JDDW 2011
  • Place of Presentation: 福岡
  • Year and Date: 2011-10-20
• [Presentation] HB-EGF-CTFの核内移行は胃癌浸潤を促進する (2011)
  • Author(s): 志村貴也,吉田道弘,福田信治,溝下 勤,片岡洋望,東山繁樹,城 卓志
  • Organizer: 第70回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2011-10-05
• [Presentation] IBD 腸管炎症に関わる炎症性サイトカインによるEGF signalを介した大腸癌細胞増殖機序 -EGF-C末端signalを標的とした新規薬剤探索- (2011)
  • Author(s): 尾関啓司、谷田諭史、溝下 勤、塚本宏延、城 卓志
  • Organizer: 第7回消化管学会総会
  • Place of Presentation: 京都
  • Year and Date: 2011-02-18
• [Presentation] IBD 腸管炎症に関わる炎症性サイトカインによるEGFシグナルを介した大腸細胞増殖機序‐HB-EGF-C末端シグナルを標的とした網羅的薬剤探索‐ (2011)
  • Author(s): 谷田 諭史、溝下 勤 、水島 隆史、城 卓志
  • Organizer: 厚生労働省「難治性炎症性腸管障害に関する調査研究」平成22年度第2回総会 免疫関連バイオマーカーの開発.
  • Place of Presentation: 東京
  • Year and Date: 2011-01-28
• [Presentation] TGFbeta induces shedding of proHB-EGF and EGFR transactivasion though ADAM activation. (2010)
  • Author(s): 海老正秀、城 卓志、神谷武、片岡洋望、久保田英嗣、溝下 勤、志村貴也、村上賢治、平田慶和、東山繁樹.
  • Organizer: 第69回 日本癌学会総会 English Workshops
  • Place of Presentation: 東京
  • Year and Date: 2010-09-24
• [Presentation] HB-EGF-C末端核移行シグナルをターゲットにした新規薬剤探索と細胞増殖抑制効果について. (2010)
  • Author(s): 尾関啓司、谷田諭史、福田信治、東山繁樹、城 卓志
  • Organizer: 第47回日本消化器免疫学会総会 一般演題
  • Place of Presentation: 大津
  • Year and Date: 2010-07-08
• [Presentation] HB-EGF-C末端核移行シグナルをターゲットにした新規薬剤探索と細胞増殖抑制効果について (2010)
  • Author(s): 尾関啓司
  • Organizer: 第47回日本消化器免疫学会総会
  • Place of Presentation: 大津プリンスホテル(滋賀県)
  • Year and Date: 2010-07-08
• [Presentation] 大腸癌増殖シグナルHB-EGF-C 末端核移行をターゲットとした新規薬剤探索
  • Author(s): 尾関啓司
  • Organizer: 第50回日本癌治療学会学術集会
  • Place of Presentation: パシフィコ横浜（神奈川）
• [Presentation] 炎症性サイトカイン(IL-8)によるHB-EGF-C末端核移行シグナルを介した大腸細胞増殖機序
  • Author(s): 尾関啓司
  • Organizer: 第40回日本潰瘍学会
  • Place of Presentation: 京王プラザホテル（東京）
• [Remarks] 名古屋市立大学大学院医学研究科消化器代謝内科学ホームページ業績集
  • URL: http://www.ncu-shotai.ac/index.html
• [Remarks] 名古屋市立大学医学研究科消化器代謝内科学 研究業績
  • URL: http://www.ncu-shotai.ac/index.html