title: Biodistribution and safety assessment of bladder cancer specific recombinant oncolytic adenovirus in subcutaneous xenografts tumor model in nude mice.
abstract: BACKGROUND
  The previous works about safety evaluation for constructed bladder tissue specific adenovirus are poorly documented. Thus, we investigated the biodistribution and body toxicity of bladder specific oncolytic adenovirus Ad-PSCAE-UPII-E1A (APU-E1A) and Ad-PSCAE-UPII-E1A-AR (APU-E1A-AR), providing meaningful information prior to embarking on human clinical trials.
  
  
  METHODS
  Conditionally replicate recombinant adenovirus (CRADs) APU-E1A, APU-EIA-AR were constructed with bladder tissue specific UroplakinII(UPII) promoter to induce the expression of Ad5E1A gene and E1A-AR fusing gene, and PSCAE was inserted at upstream of promoter to enhance the function of promoter. Based on the cytopathic and anti-tumor effect of bladder cancer, these CRADs were intratumorally injected into subcutaneous xenografts tumor in nude mice. We then determined the toxicity through general health and behavioral assessment, hepatic and hematological toxicity evaluation, macroscopic and microscopic postmortem analyses. The spread of the transgene E1A of adenovirus was detected with RT-PCR and Western blot. Virus replication and distribution were examined with APU-LUC administration and Luciferase Assay.
  
  
  RESULTS
  General assessment and body weight of the animals did not reveal any alteration in general behavior. The hematological alterations of groups which were injected with 5x10(8) pfu or higher dose (5x10(9) pfu) of APU-E1A and APU-E1A-AR showed no difference in comparison with PBS group, and only slight increased transaminases in contrast to PBS group at 5x10(9) pfu of APU-E1A and APU-E1A-AR were observed. E1A transgene did not disseminate to organs outside of xenograft tumor. Virus replication was not detected in other organs beside tumor according to Luciferase Assay.
  
  
  CONCLUSIONS
  Our study showed that recombinant adenovirus APU-E1A-AR and APU-E1A appear safe with 5x10(7) pfu and 5x10(8) pfu intratumorally injection in mice, without any discernable effects on general health and behavior.
authors.affiliation: School of Life Sciences in Lanzhou University, Lanzhou, Gansu Province, China. erywzp@lzu.edu.cn
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Wang
authors.forename: Hongwei
authors.initials: H
authors.lastname: Tian
authors.forename: Meijiao
authors.initials: M
authors.lastname: Qi
authors.forename: Zhenxing
authors.initials: Z
authors.lastname: Zhai
authors.forename: Shuwen
authors.initials: S
authors.lastname: Li
authors.forename: Renju
authors.initials: R
authors.lastname: Li
authors.forename: Hongjuan
authors.initials: H
authors.lastname: Zhang
authors.forename: Wenyun
authors.initials: W
authors.lastname: Wang
authors.forename: Shenjun
authors.initials: S
authors.lastname: Fu
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Lu
authors.forename: Ronald
authors.initials: R
authors.lastname: Rodriguez
authors.forename: Yinglu
authors.initials: Y
authors.lastname: Guo
authors.forename: Liqun
authors.initials: L
authors.lastname: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003588:Cytopathogenic Effect, Viral; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D050130:Oncolytic Virotherapy / Q000379:methods; D050504:Oncolytic Viruses / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D060170:Uroplakin II / Q000235:genetics; D023041:Xenograft Model Antitumor Assays / Q000379:methods*
chemical_list: D017500:Adenovirus E1A Proteins; D011993:Recombinant Fusion Proteins; D060170:Uroplakin II