Midkine-Promoter based Conditionally Replicative Adenovirus expressing siRNA against EGFR for Head and Neck Cancer

• Project/Area Number: 16H05481
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Otorhinolaryngology
• Research Institution: Kobe University
• Principal Investigator: Nibu Ken-ichi 神戸大学, 医学研究科, 教授 (20251283)
• Co-Investigator(Kenkyū-buntansha): 大月 直樹 神戸大学, 医学研究科, 准教授 (40343264) ; 上原 奈津美 神戸大学, 医学部附属病院, 助教 (40570502)
• Research Collaborator: SHIRAKAWA toshiro
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2018: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2016: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 頭頸部癌 / Midkine / EGFR / siRNA / 制限増殖型アデノウイルス / 遺伝子治療 / アデノウイルスベクター / 増殖制限型アデノウイルス / MIDKINE / 癌 / 遺伝子 / ウイルス / ウィルス

Outline of Final Research Achievements

Background: Epidermal growth factor receptor (EGFR) is overexpressed in head and neck squamous cell carcinomas (HNSCCs). Midkine expression is restricted in adult tissues, but is increased in several malignant tumors, including HNSCCs. Here we evaluated the antitumor effect of Midkine-promoter based conditionally replicative adenovirus expressing siRNA against EGFR for targeting HNSCCs expressing Midkine. Methods: A conditionally replicative adenovirus vector controlled by the Midkine promoter, Ad-MK-siEGFR, was generated by integrating gene-expressing siRNA against EGFR. Antitumor effect of Ad-MK-siEGFR was tested in vitro using established HNSCC cell line, T891 with strong Midkine expression. Results: Expression of EGFR in T891 infected with Ad-MKsiEGFR was significantly lower than that of T891 infected with control. Cytotoxicity assays showed significant growth suppression of Ad-MK-siEGFR in T891 cells.

Academic Significance and Societal Importance of the Research Achievements

今回我々はMidkineプロモーターをつけて腫瘍特異的に増殖する制限増殖型アデノウイルスAd-MKを作成し、さらにこのアデノウイルスに頭頸部扁平上皮癌に高発現している上皮成長因子受容体EGFRに対するsiRNAの遺伝子を組み込んだAd-MKsiEGFRを作成した。in vitroではAd-MK、Ad-MKsiEGFRともに頭頸部癌細胞の増殖を著明に抑制できた。Ad-MKsiEGFRのsiRNAの効果もmRNAおよび蛋白レベルで確認でき、２重の抗腫瘍効果が確認された。本方法は、再発率が高く予後不良である進行性頭頸部癌の新たなより効果的な治療の開発に向けた基礎的な研究となった。

Research Products

• [Journal Article] Clinical impact of a cytological screening system using cyclin D1 immunostaining and genomic analysis for the diagnosis of thyroid nodules (2019)
  • Author(s): Teshima M, Tokita K, Ryo E, Matsumoto F, Kondo M, Ikegami Y, Shinomiya H, Otsuki N, Hiraoka N, Nibu KI, Yoshimoto S, Mori T
  • Journal Title: BMC Cancer. Volume: 19 Issue: 1 Pages: 245-245
  • DOI: 10.1186/s12885-019-5452-4
  • Peer Reviewed / Open Access
• [Journal Article] Overexpression of SOCS3 mediated by adenovirus vector in mouse and human castration-resistant prostate cancer cells increases the sensitivity to NK cells in vitro and in vivo (2019)
  • Author(s): Yoneda T, Kunimura N, Kitagawa K, Fukui Y, Saito H, Narikiyo K, Ishiko M, Otsuki N, Nibu KI, Fujisawa M, Serada S, Naka T, Shirakawa T.
  • Journal Title: Cancer Gene Ther Volume: in print
  • NAID: 120006951631
  • Peer Reviewed
• [Journal Article] Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA combined with cisplatin inhibits cellular growth and induces apoptosis in HPV-positive head and neck cancer cells (2018)
  • Author(s): Kojima Y, Otsuki N, Kubo M, Kitamoto J, Takata E, Saito H, Kosaka K, Morishita N, Uehara N, Shirakawa T, Nibu KI
  • Journal Title: Cancer Gene Ther Volume: 25 Issue: 9-10 Pages: 274-283
  • DOI: 10.1038/s41417-018-0024-3
  • Peer Reviewed
• [Journal Article] Mutation analysis of the EGFR pathway genes, EGFR, RAS, PIK3CA, BRAF, and AKT1, in salivary gland adenoid cystic carcinoma. (2018)
  • Author(s): Saida K, Murase T, Ito M, Fujii K, Takino H, Masaki A, Kawakita D, Ijichi K, Tada Y, Kusafuka K,Iida Y, Onitsuka T, Yatabe Y, Hanai N, Hasegawa Y
  • Journal Title: Oncotarget Volume: 9 Issue: 24 Pages: 17043-17055
  • DOI: 10.18632/oncotarget.24818
  • Peer Reviewed / Open Access
• [Journal Article] Epigenetic down-regulation of SOX2 is an independent poor prognostic factor for hypopharyngeal cancers (2018)
  • Author(s): Avincsal Mehmet Ozgur、Jimbo Naoe、Fujikura Kohei、Shinomiya Hirotaka、Otsuki Naoki、Morimoto Koichi、Furukawa Tatsuya、Morita Naruhiko、Maehara Ritsuko、Itoh Tomoo、Nibu Ken-ichi、Zen Yoh
  • Journal Title: Histopathology Volume: 72 Issue: 5 Pages: 826-837
  • DOI: 10.1111/his.13436
  • Peer Reviewed
• [Journal Article] The Role of Molecular Testing in the Differential Diagnosis of Salivary Gland Carcinomas (2018)
  • Author(s): Sk?lov? Alena、Stenman G?ran、Simpson Roderick H.W.、Hellquist Henrik、Slouka David、Svoboda Tomas、Bishop Justin A.、Hunt Jennifer L.、Nibu Ken-Ichi、Rinaldo Alessandra、Vander Poorten Vincent、Devaney Kenneth O.、Steiner Petr、Ferlito Alfio
  • Journal Title: The American Journal of Surgical Pathology Volume: 42 Issue: 2 Pages: e11-e27
  • DOI: 10.1097/pas.0000000000000980
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Impact of alcohol dehydrogenase-aldehyde dehydrogenase polymorphism on clinical outcome in patients with hypopharyngeal cancer (2017)
  • Author(s): Avin?sal Mehmet Ozgur、Shinomiya Hirotaka、Teshima Masanori、Kubo Mie、Otsuki Naoki、Kyota Naomi、Sasaki Ryohei、Zen Yoh、Nibu Ken-ichi
  • Journal Title: Head & Neck Volume: 40 Issue: 4 Pages: 770-777
  • DOI: 10.1002/hed.25050
  • Peer Reviewed
• [Journal Article] Prognostic value of ALDH2 polymorphism for patients with oropharyngeal cancer in a Japanese population (2017)
  • Author(s): Shinomiya Hirotaka、Shinomiya Hitomi、Kubo Mie、Saito Yuki、Yoshida Masafumi、Ando Mizuo、Teshima Masanori、Otsuki Naoki、Kiyota Naomi、Sasaki Ryohei、Nibu Ken-ichi
  • Journal Title: PLOS ONE Volume: 12 Issue: 12 Pages: e0187992-e0187992
  • DOI: 10.1371/journal.pone.0187992
  • NAID: 120006380399
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Organ preservation with chemoradiation in advanced laryngeal cancer: The problem of generalizing results from randomized controlled trials (2017)
  • Author(s): Sanabria Alvaro、Chaves Aline L.F.、Kowalski Luiz P.、Wolf Gregory T.、Saba Nabil F.、Forastiere Arlene A.、Beitler Jonathan J.、Nibu Ken-ichi、Bradford Carol R.、Su?rez Carlos、Rodrigo Juan P.、Strojan Primo?、Rinaldo Alessandra、de Bree Remco、Haigentz Missak、Takes Robert P.、Ferlito Alfio
  • Journal Title: Auris Nasus Larynx Volume: 44 Issue: 1 Pages: 18-25
  • DOI: 10.1016/j.anl.2016.06.005
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Expression of amphiregulin in mucoepidermoid carcinoma of the major salivary glands: a molecular and clinicopathological study (2016)
  • Author(s): Shinomiya H, Ito Y, Kubo M, Yonezawa K, Otsuki N, Iwae S, Inagaki H, Nibu KI
  • Journal Title: Hum Pathol Volume: 57 Pages: 37-44
  • DOI: 10.1016/j.humpath.2016.06.016
  • Peer Reviewed