title: Quantifying the activity of adenoviral E1A CR2 deletion mutants using renilla luciferase bioluminescence and 3'-deoxy-3'-[18F]fluorothymidine positron emission tomography imaging.
abstract: The adenoviral E1A CR2 mutant dl922-947 has potent activity in ovarian cancer. We have used Renilla luciferase bioluminescence imaging to monitor viral E1A expression and replication and [18F]fluorothymidine positron emission tomography ([18F]FLT-PET) to quantify the activity of dl922-947 in vivo. We created dlCR2 Ren, with the same E1A CR2 deletion as dl922-947 and the luciferase gene from Renilla reniformis downstream of E1. Light emitted from s.c. and i.p. IGROV1 ovarian carcinoma xenografts was measured following treatment with dlCR2 Ren. Mice bearing s.c. IGROV1 xenografts were injected with 2.96 to 3.7 MBq of [18F]FLT 48 and 168 hours following i.t. injection of dl922-947 or control virus Ad LM-X. The presence of Renilla luciferase in dlCR2 Ren did not reduce in vitro nor in vivo potency compared with dl922-947. Light emission correlated closely with E1A expression in vitro and peaked 48 hours after dlCR2 Ren injection in both s.c. and i.p. IGROV1 xenografts. It diminished by 168 hours in s.c. tumors but persisted for at least 2 weeks in i.p. models. Normalized tumor [18F]FLT uptake at 60 minutes (NUV60), fractional retention, and area under radioactivity curve all decreased marginally 48 hours after dl922-947 treatment and significantly at 168 hours compared with controls. There was a close linear correlation between NUV60 and both tumor proliferation (Ki67 labeling index) and thymidine kinase 1 expression. Renilla luciferase bioluminescence and [18F]FLT-PET imaging are capable of quantifying the activity and effectiveness of E1A CR2-deleted adenoviral mutants in ovarian cancer.
authors.affiliation: Molecular Therapy and PET Oncology Research Group, Clinical Sciences Centre, Imperial College School of Medicine, London, United Kingdom.
authors.forename: Julius
authors.initials: J
authors.lastname: Leyton
authors.forename: Michelle
authors.initials: M
authors.lastname: Lockley
authors.forename: Joeri L
authors.initials: JL
authors.lastname: Aerts
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Baird
authors.forename: Eric O
authors.initials: EO
authors.lastname: Aboagye
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Lemoine
authors.forename: Iain A
authors.initials: IA
authors.lastname: McNeish
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D017500:Adenovirus E1A Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D015224:Dideoxynucleosides; D005260:Female; D005462:Fluorine Radioisotopes; D017353:Gene Deletion; D006801:Humans; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D051379:Mice; D008807:Mice, Inbred BALB C; D050130:Oncolytic Virotherapy / Q000379:methods; D010051:Ovarian Neoplasms / Q000000981:diagnostic imaging / Q000628:therapy / Q000821:virology*; D049268:Positron-Emission Tomography / Q000379:methods; D019275:Radiopharmaceuticals; D014779:Virus Replication / Q000502:physiology; D023041:Xenograft Model Antitumor Assays
chemical_list: D017500:Adenovirus E1A Proteins; D015224:Dideoxynucleosides; D005462:Fluorine Radioisotopes; D019275:Radiopharmaceuticals; D049410:Luciferases, Renilla; C002854:alovudine