title: Microvascular free tissue transfer for gene delivery: in vivo evaluation of different routes of plasmid and adenoviral delivery.
abstract: Transfer of healthy autologous tissue as a microvascular free flap facilitates reconstruction during ablative cancer surgery. In addition to filling surgical defects, free flaps might concentrate viral vectors at the tumour bed and mediate local therapeutic effects. We evaluated the magnitude, topography and duration of luciferase gene expression after plasmid and adenoviral delivery in rat superficial inferior epigastric (SIE) flaps. For plasmid delivery, luciferase expression was significantly increased by all transduction routes (topical, intraflap injection, intravascular) (P<0.01) at day 1, but not at day 7. The spread of luciferase expression was significantly different between the 4 groups at 1 day (P=0.026) and was greatest for flaps transduced by intravascular injection. For adenoviral transduction, total radiance was significantly different between the transduced groups at 1, 14 and 28 days (P<0.05 for all comparisons). The highest levels of radiance were seen in the intravascular group. There was a statistically significant difference in the spread of light emission between the 3 groups at 1 (P=0.009) and 14 (P=0.013) days, but this was no longer evident at 28 days. Intravascular adenoviral delivery yields high-level, diffuse and durable gene expression in rat SIE flaps and is suitable for examination in therapeutic models.
authors.affiliation: Chester Beatty Laboratories, The Institute of Cancer Research, London, UK.
authors.forename: V K
authors.initials: VK
authors.lastname: Agrawal
authors.forename: K M
authors.initials: KM
authors.lastname: Copeland
authors.forename: Y
authors.initials: Y
authors.lastname: Barbachano
authors.forename: A
authors.initials: A
authors.lastname: Rahim
authors.forename: R
authors.initials: R
authors.lastname: Seth
authors.forename: C L
authors.initials: CL
authors.lastname: White
authors.forename: M
authors.initials: M
authors.lastname: Hingorani
authors.forename: C M
authors.initials: CM
authors.lastname: Nutting
authors.forename: M
authors.initials: M
authors.lastname: Kelly
authors.forename: P
authors.initials: P
authors.lastname: Harris
authors.forename: H
authors.initials: H
authors.lastname: Pandha
authors.forename: A A
authors.initials: AA
authors.lastname: Melcher
authors.forename: R G
authors.initials: RG
authors.lastname: Vile
authors.forename: C
authors.initials: C
authors.lastname: Porter
authors.forename: K J
authors.initials: KJ
authors.lastname: Harrington
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D007267:Injections; D007763:Lac Operon; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008297:Male; D023421:Models, Animal; D010957:Plasmids / Q000494:pharmacology*; D051381:Rats; D011916:Rats, Inbred F344; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013524:Surgical Flaps; D014018:Tissue Distribution; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D008156:Luciferases