PURPOSE: Telomerase reverse transcriptase (hTERT) is the key determinant of telomerase activity and plays a crucial role in cellular immortalization and oncogenesis. It will be a promising target for cancer gene therapy. We constructed a novel replicative adenovirus CNHK300 in which hTERT promoter with three extra E-boxes downstream of the promoter was introduced and used to regulate adenoviral E1a gene, and studied its properties of selective replication in cancer cells and antitumoral activity. METHODS: Luciferase assay was used to detect hTERT promoter activity. The selective replication of CNHK300 in cancer cells was investigated by E1a Western blot and green fluorescent protein (GFP) reporter gene assay. The antitumoral activity of CNHK300 and its toxicity were measured on animal models. RESULTS: Luciferase assay showed that introducing extra E-boxes downstream of hTERT promoter is beneficial to decreasing the promoter activity in normal cells without affecting its strong activity in cancer cells. Experiments in vitro and in vivo demonstrated that CNHK300 can selectively target to hTERT-positive cancer cells and replicate in them, resulting in oncolytic or antitumoral effect. CNHK300 is superior to ONYX-015 in terms of selective replication and oncolytic or antitumoral effect. The toxicity assay showed no signs of toxicity to liver cells even at the higher dosage of CNHK300 in vivo. CONCLUSION: The hTERT promoter-controlled, replication-competent adenovirus CNHK300 is a promising system for targeted cancer gene therapy.
PURPOSE: Telomerase reverse transcriptase (hTERT) is the key determinant of telomerase activity and plays a crucial role in cellular immortalization and oncogenesis. It will be a promising target for cancer gene therapy. We constructed a novel replicative adenovirus CNHK300 in which hTERT promoter with three extra E-boxes downstream of the promoter was introduced and used to regulate adenoviral E1a gene, and studied its properties of selective replication in cancer cells and antitumoral activity. METHODS: Luciferase assay was used to detect hTERT promoter activity. The selective replication of CNHK300 in cancer cells was investigated by E1a Western blot and green fluorescent protein (GFP) reporter gene assay. The antitumoral activity of CNHK300 and its toxicity were measured on animal models. RESULTS: Luciferase assay showed that introducing extra E-boxes downstream of hTERT promoter is beneficial to decreasing the promoter activity in normal cells without affecting its strong activity in cancer cells. Experiments in vitro and in vivo demonstrated that CNHK300 can selectively target to hTERT-positive cancer cells and replicate in them, resulting in oncolytic or antitumoral effect. CNHK300 is superior to ONYX-015 in terms of selective replication and oncolytic or antitumoral effect. The toxicity assay showed no signs of toxicity to liver cells even at the higher dosage of CNHK300 in vivo. CONCLUSION: The hTERT promoter-controlled, replication-competent adenovirus CNHK300 is a promising system for targeted cancer gene therapy.
Authors: Leisa Johnson; Annie Shen; Larry Boyle; John Kunich; Kusum Pandey; Marilyn Lemmon; Terry Hermiston; Marty Giedlin; Frank McCormick; Ali Fattaey Journal: Cancer Cell Date: 2002-05 Impact factor: 31.743
Authors: M Ohashi; F Kanai; K Tateishi; H Taniguchi; P A Marignani; Y Yoshida; Y Shiratori; H Hamada; M Omata Journal: Biochem Biophys Res Commun Date: 2001-03-30 Impact factor: 3.575
Authors: D Xu; Q Wang; A Gruber; M Björkholm; Z Chen; A Zaid; G Selivanova; C Peterson; K G Wiman; P Pisa Journal: Oncogene Date: 2000-10-26 Impact factor: 9.867
Authors: X Y Pan; X J Liu; J Li; S J Zhen; D X Liu; Q Feng; W X Zhao; Y Luo; Y L Zhang; H W Li; J L Yang Journal: Gene Ther Date: 2016-12-22 Impact factor: 5.250
Authors: S Bortolanza; C Qian; M G Kramer; C Gomar; J Prieto; F Farinati; R Hernandez-Alcoceba Journal: J Mol Med (Berl) Date: 2005-06-23 Impact factor: 4.599
Authors: Juming Ma; J Ma; Xiaoping He; X He; Weiguo Wang; W Wang; Yao Huang; Y Huang; Lin Chen; L Chen; Wenming Cong; W Cong; Jianzhong Gu; J Gu; Huizhen Hu; H Hu; Jianguo Shi; J Shi; Linfang Li; L Li; Changqing Su; C Su Journal: Dig Dis Sci Date: 2008-11-26 Impact factor: 3.199
Authors: Maria Rajecki; Mirkka Sarparanta; Tanja Hakkarainen; Mikko Tenhunen; Iulia Diaconu; Venla Kuhmonen; Kalevi Kairemo; Anna Kanerva; Anu J Airaksinen; Akseli Hemminki Journal: PLoS One Date: 2012-03-07 Impact factor: 3.240