Molecular PET imaging of HSV1-tk reporter gene expression using [18F]FEAU.
Non-invasive imaging of transgene expression requires the appropriate combination of a reporter gene and a reporter probe. [18F]FEAU positron emission tomography (PET) is used for the assessment of herpes simplex virus type-1 thymidine kinase gene expression. Hybrid AAV phage (termed AAVP) can be adapted to transduce mammalian cells by targeting to a specific receptor. We evaluated a targeted AAVP vector using [18F]FEAU PET. This protocol describes [18F]FEAU production and dosing, micro-PET imaging and image analysis. 2-Deoxy-2-trifluoromethanesulfonyl-1,3,5-tri-O-benzoyl-alpha-D-ribofuranose is radio-fluorinated, converted into its 1-bromo derivative and coupled with protected 5-ethyl uracil. The coupled product is hydrolyzed and purified using HPLC. Tumor-bearing animals targeted with either retroviral or AAVP vectors are anesthetized and injected with [18F]FEAU (0.1 mCi per mouse); this is followed 2 h after injection by imaging on a micro-PET. Production of [18F]FEAU requires approximately 3.5 h from the end of bombardment. PET imaging studies require 2-3 h (depending on the number of animals) after synthesis of [18F]FEAU.
Soghomonyan
Suren
S
The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77030, USA.
Hajitou
Amin
A
Rangel
Roberto
R
Trepel
Martin
M
Pasqualini
Renata
R
Arap
Wadih
W
Gelovani
Juri G
JG
Alauddin
Mian M
MM
D000818:Animals; D001086:Arabinofuranosyluracil / Q000031:analogs & derivatives; D045744:Cell Line, Tumor; D005462:Fluorine Radioisotopes; D020869:Gene Expression Profiling / Q000379:methods*; D018390:Gene Targeting; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D051379:Mice; D015394:Molecular Structure; D049268:Positron-Emission Tomography / Q000379:methods*; D013937:Thymidine Kinase / Q000378:metabolism*; D019076:Transgenes / Q000235:genetics
D005462:Fluorine Radioisotopes; D001086:Arabinofuranosyluracil; C054103:2'-fluoro-5-ethylarabinosyluracil; D013937:Thymidine Kinase