start-ver=1.4 cd-journal=joma no-vol=119 cd-vols= no-issue=10 article-no= start-page=3172 end-page=3181 dt-received= dt-revised= dt-accepted= dt-pub-year=2009 dt-pub=20091001 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=A simple biological imaging system for detecting viable human circulating tumor cells en-subtitle= kn-subtitle= en-abstract= kn-abstract=The presence of circulating tumor cells (CTCs) in the peripheral blood is associated with short survival, making the detection of CTCs clinically useful as a prognostic factor of disease outcome and/or a surrogate marker of treatment response. Recent technical advances in immunocytometric analysis and quantitative real-time PCR have made it possible to detect a few CTCs in the blood; however, there is no sensitive assay to specifically detect viable CTCs. Here, we report what we believe to be a new approach to visually detect live human CTCs among millions of peripheral blood leukocytes, using a telomerase-specific replication-selective adenovirus expressing GFP. First, we constructed a GFP-expressing attenuated adenovirus, in which the telomerase promoter regulates viral replication (OBP-401; TelomeScan). We then used OBP-401 to establish a simple ex vivo method that was able to detect viable human CTCs in the peripheral blood. The detection method involved a 3-step procedure, including the lysis of rbc, the subsequent addition of OBP-401 to the cell pellets, and an automated scan using fluorescence microscopy. OBP-401 infection increased the signal-to-background ratio as a tumor-specific probe, because the fluorescent signal was amplified only in viable, infected human tumor cells, by viral replication. This GFP-expressing virus-based method is remarkably simple and allows precise enumeration of CTCs. en-copyright= kn-copyright= en-aut-name=KojimaToru en-aut-sei=Kojima en-aut-mei=Toru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HashimotoYuuri en-aut-sei=Hashimoto en-aut-mei=Yuuri kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=WatanabeYuichi en-aut-sei=Watanabe en-aut-mei=Yuichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KagawaShunsuke en-aut-sei=Kagawa en-aut-mei=Shunsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=UnoFutoshi en-aut-sei=Uno en-aut-mei=Futoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=KurodaShinji en-aut-sei=Kuroda en-aut-mei=Shinji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=TazawaHiroshi en-aut-sei=Tazawa en-aut-mei=Hiroshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=KyoSatoru en-aut-sei=Kyo en-aut-mei=Satoru kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=MizuguchiHiroyuki en-aut-sei=Mizuguchi en-aut-mei=Hiroyuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=UrataYasuo en-aut-sei=Urata en-aut-mei=Yasuo kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=TanakaNoriaki en-aut-sei=Tanaka en-aut-mei=Noriaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= en-aut-name=FujiwaraToshiyoshi en-aut-sei=Fujiwara en-aut-mei=Toshiyoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=12 ORCID= affil-num=1 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=2 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=3 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=4 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=5 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=6 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=7 en-affil= kn-affil=Center for Gene and Cell Therapy, Okayama University Hospital affil-num=8 en-affil= kn-affil=Department of Obstetrics and Gynecology, Kanazawa University School of Medicine affil-num=9 en-affil= kn-affil=Department of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University affil-num=10 en-affil= kn-affil=Oncolys BioPharma Inc. affil-num=11 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences affil-num=12 en-affil= kn-affil=Division of Surgical Oncology, Department of Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences END start-ver=1.4 cd-journal=joma no-vol=124 cd-vols= no-issue=2 article-no= start-page=105 end-page=110 dt-received= dt-revised= dt-accepted= dt-pub-year=2012 dt-pub=20120801 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Preclinical evaluation of telomerase-specific oncolytic virotherapy for human bone and soft tissue sarcomas kn-title=テロメラーゼ依存性腫瘍融解ウイルス療法の骨・軟部肉腫に対する前臨床的検討 en-subtitle= kn-subtitle= en-abstract= kn-abstract=骨・軟部肉腫は, 一部に治療抵抗性で予後の悪い症例が存在するため, 新たな治療法の確立が重要な課題である.　我々は, 5型アデノウイルスを基本骨格として, テロメラーゼ活性に依存して増殖する腫瘍融解ウイルス（OBP-301）や, coxsackie and adenovirus receptor（CAR）陰性の腫瘍細胞に感染するファイバー改変型ウイルス（OBP-405）を用い, 骨・軟部肉腫細胞に対する抗腫瘍効果を検討した.　 　14種類の骨・軟部肉腫細胞株に対してOBP-301の細胞障害活性を検討し, 12種類の細胞株でOBP-301に感受性を認めた.　また, OBP-301の細胞障害活性はCARの発現と相関していた.　さらに, テロメラーゼ活性の低い細胞に対しても, 5型アデノウイルスの複製に必須のE1Aによりテロメラーゼ活性の増強効果がおこり, 強い抗腫瘍活性を示すことを明らかにした.　次に, 骨肉腫脛骨同所性移植動物モデルを作成しOBP-301を投与したところ, OBP-301投与群では対象群と比べて有意に腫瘍増殖を抑制した.　最後に, OBP-301に感受性を認めなかったCAR陰性細胞株に対してOBP-405を用いて検討し, OBP-405が有効に作用することを確認した.　 　OBP-301やOBP-405を用いたウイルス療法は, 骨・軟部肉腫に対する新たな治療法となる可能性がある.　 en-copyright= kn-copyright= en-aut-name=SasakiTsuyoshi en-aut-sei=Sasaki en-aut-mei=Tsuyoshi kn-aut-name=佐々木剛 kn-aut-sei=佐々木 kn-aut-mei=剛 aut-affil-num=1 ORCID= en-aut-name=TazawaHiroshi en-aut-sei=Tazawa en-aut-mei=Hiroshi kn-aut-name=田澤大 kn-aut-sei=田澤 kn-aut-mei=大 aut-affil-num=2 ORCID= en-aut-name=HaseiJo en-aut-sei=Hasei en-aut-mei=Jo kn-aut-name=長谷井嬢 kn-aut-sei=長谷井 kn-aut-mei=嬢 aut-affil-num=3 ORCID= en-aut-name=KunisadaToshiyuki en-aut-sei=Kunisada en-aut-mei=Toshiyuki kn-aut-name=国定俊之 kn-aut-sei=国定 kn-aut-mei=俊之 aut-affil-num=4 ORCID= en-aut-name=YoshidaAki en-aut-sei=Yoshida en-aut-mei=Aki kn-aut-name=吉田晶 kn-aut-sei=吉田 kn-aut-mei=晶 aut-affil-num=5 ORCID= en-aut-name=HashimotoYuuri en-aut-sei=Hashimoto en-aut-mei=Yuuri kn-aut-name=橋本悠里 kn-aut-sei=橋本 kn-aut-mei=悠里 aut-affil-num=6 ORCID= en-aut-name=YanoShuya en-aut-sei=Yano en-aut-mei=Shuya kn-aut-name=矢野修也 kn-aut-sei=矢野 kn-aut-mei=修也 aut-affil-num=7 ORCID= en-aut-name=YoshidaRyosuke en-aut-sei=Yoshida en-aut-mei=Ryosuke kn-aut-name=吉田亮介 kn-aut-sei=吉田 kn-aut-mei=亮介 aut-affil-num=8 ORCID= en-aut-name=UnoFutoshi en-aut-sei=Uno en-aut-mei=Futoshi kn-aut-name=宇野太 kn-aut-sei=宇野 kn-aut-mei=太 aut-affil-num=9 ORCID= en-aut-name=KagawaShunsuke en-aut-sei=Kagawa en-aut-mei=Shunsuke kn-aut-name=香川俊輔 kn-aut-sei=香川 kn-aut-mei=俊輔 aut-affil-num=10 ORCID= en-aut-name=MorimotoYuki en-aut-sei=Morimoto en-aut-mei=Yuki kn-aut-name=森本裕樹 kn-aut-sei=森本 kn-aut-mei=裕樹 aut-affil-num=11 ORCID= en-aut-name=UrataYasuo en-aut-sei=Urata en-aut-mei=Yasuo kn-aut-name=浦田泰生 kn-aut-sei=浦田 kn-aut-mei=泰生 aut-affil-num=12 ORCID= en-aut-name=FujiwaraToshiyoshi en-aut-sei=Fujiwara en-aut-mei=Toshiyoshi kn-aut-name=藤原俊義 kn-aut-sei=藤原 kn-aut-mei=俊義 aut-affil-num=13 ORCID= en-aut-name=OzakiToshifumi en-aut-sei=Ozaki en-aut-mei=Toshifumi kn-aut-name=尾﨑敏文 kn-aut-sei=尾﨑 kn-aut-mei=敏文 aut-affil-num=14 ORCID= affil-num=1 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 affil-num=2 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=3 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 affil-num=4 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 affil-num=5 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 affil-num=6 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=7 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=8 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=9 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=10 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=11 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 affil-num=12 en-affil= kn-affil=オンコリスバイオファーマ affil-num=13 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=14 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　整形外科学 en-keyword=アデノウイルス kn-keyword=アデノウイルス en-keyword=肉腫 kn-keyword=肉腫 en-keyword=ALT kn-keyword=ALT en-keyword=ヒトテロメラーゼ逆転写酵素 kn-keyword=ヒトテロメラーゼ逆転写酵素 END start-ver=1.4 cd-journal=joma no-vol=123 cd-vols= no-issue=2 article-no= start-page=103 end-page=109 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=20110801 dt-online= en-article= kn-article= en-subject= kn-subject= en-title=Radiosensitization by telomerase-dependent oncolytic adenovirus kn-title=テロメラーゼ依存的腫瘍融解アデノウイルス製剤による 放射線感受性増強作用 en-subtitle= kn-subtitle= en-abstract= kn-abstract=DNA修復機能阻害は放射線感受性を増強させるため，DNA修復に関与する因子の阻害剤は放射線増感剤となり得る．我々の開発したテロメラーゼ依存的腫瘍融解アデノウイルス製剤OBP-301（テロメライシン）は，アデノウイルスE1B55kDaタンパクを介して細胞のDNA修復に重要な役割を果たすMRN複合体（Mre11，Rad50，NBS1）を分解する機能を有する．このMRN複合体の分解によりATM（ataxia-telangiectasia mutated）の活性化が抑制され結果的にDNA修復機構が阻害される．我々はOBP-301と放射線との併用が強力な相乗効果を生み出すことをマウスの皮下腫瘍モデルおよび食道癌同所性モデルにおいて証明した．これらの結果はOBP-301が将来有望な放射線増感剤となり得ることだけでなく，E1B55kDaタンパクを産生する腫瘍融解アデノウイルス製剤と放射線との併用が悪性腫瘍に対する有力な治療戦略となり得ることを示す． en-copyright= kn-copyright= en-aut-name=KurodaShinji en-aut-sei=Kuroda en-aut-mei=Shinji kn-aut-name=黒田新士 kn-aut-sei=黒田 kn-aut-mei=新士 aut-affil-num=1 ORCID= en-aut-name=FujiwaraToshiya en-aut-sei=Fujiwara en-aut-mei=Toshiya kn-aut-name=藤原俊哉 kn-aut-sei=藤原 kn-aut-mei=俊哉 aut-affil-num=2 ORCID= en-aut-name=ShirakawaYasuhiro en-aut-sei=Shirakawa en-aut-mei=Yasuhiro kn-aut-name=白川靖博 kn-aut-sei=白川 kn-aut-mei=靖博 aut-affil-num=3 ORCID= en-aut-name=YamasakiYasumoto en-aut-sei=Yamasaki en-aut-mei=Yasumoto kn-aut-name=山崎泰源 kn-aut-sei=山崎 kn-aut-mei=泰源 aut-affil-num=4 ORCID= en-aut-name=YanoSyuya en-aut-sei=Yano en-aut-mei=Syuya kn-aut-name=矢野修也 kn-aut-sei=矢野 kn-aut-mei=修也 aut-affil-num=5 ORCID= en-aut-name=UnoFutoshi en-aut-sei=Uno en-aut-mei=Futoshi kn-aut-name=宇野太 kn-aut-sei=宇野 kn-aut-mei=太 aut-affil-num=6 ORCID= en-aut-name=TazawaHiroshi en-aut-sei=Tazawa en-aut-mei=Hiroshi kn-aut-name=田澤大 kn-aut-sei=田澤 kn-aut-mei=大 aut-affil-num=7 ORCID= en-aut-name=HashimotoYuuri en-aut-sei=Hashimoto en-aut-mei=Yuuri kn-aut-name=橋本悠里 kn-aut-sei=橋本 kn-aut-mei=悠里 aut-affil-num=8 ORCID= en-aut-name=WatanabeYuichi en-aut-sei=Watanabe en-aut-mei=Yuichi kn-aut-name=渡辺雄一 kn-aut-sei=渡辺 kn-aut-mei=雄一 aut-affil-num=9 ORCID= en-aut-name=NomaKazuhiro en-aut-sei=Noma en-aut-mei=Kazuhiro kn-aut-name=野間和広 kn-aut-sei=野間 kn-aut-mei=和広 aut-affil-num=10 ORCID= en-aut-name=UrataYasuo en-aut-sei=Urata en-aut-mei=Yasuo kn-aut-name=浦田泰生 kn-aut-sei=浦田 kn-aut-mei=泰生 aut-affil-num=11 ORCID= en-aut-name=KagawaShunsuke en-aut-sei=Kagawa en-aut-mei=Shunsuke kn-aut-name=香川俊輔 kn-aut-sei=香川 kn-aut-mei=俊輔 aut-affil-num=12 ORCID= en-aut-name=FujiwaraToshiyoshi en-aut-sei=Fujiwara en-aut-mei=Toshiyoshi kn-aut-name=藤原俊義 kn-aut-sei=藤原 kn-aut-mei=俊義 aut-affil-num=13 ORCID= affil-num=1 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=2 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=3 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=4 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=5 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=6 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=7 en-affil= kn-affil=岡山大学病院　遺伝子・細胞治療センター affil-num=8 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=9 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=10 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=11 en-affil= kn-affil=オンコリスバイオファーマ株式会社 affil-num=12 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 affil-num=13 en-affil= kn-affil=岡山大学大学院医歯薬学総合研究科　消化器外科学 en-keyword=アデノウイルス kn-keyword=アデノウイルス en-keyword=E1B55kDa kn-keyword=E1B55kDa en-keyword=MRN複合体 kn-keyword=MRN複合体 en-keyword=DNA修復 kn-keyword=DNA修復 en-keyword=放射線感受性 kn-keyword=放射線感受性 END