1. Volume 15, Issue 5, Pages 921-929 (May 2007)
Imaging the Modulation of Adenoviral Kinetics and Biodistribution for Cancer Gene Therapy 
Joseph D Mocanu, Kenneth W Yip, Nehad M Alajez, Wei Shi, Jian-Hua Li, Sarah J Lunt, Eduardo H Moriyama, Brian C Wilson, Michael Milosevic, Kwok-Wai Lo, Nico van van Rooijen, Pierre Busson, Carlo Bastianutto, Fei-Fei Liu 
Molecular Therapy 
Volume 15, Issue 5, Pages (May 2007)
DOI: /mt.sj
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Schema of the PDC312-based adenoviral constructs. (a) PDC312/SV40luc, (b) PDC312/oriP.luc, and (c) PDC312/oriP.E1A.oriP.luc.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
In vitro plasmid-based and viral-based expression, and toxicity over time. (a) Normalized luciferase activity levels of PDC312/SV40luc, PDC312/oriP.luc, and PDC312/oriP.E1A.oriP.luc 24 and 48 hours after transfection into C666-1 cells; (b) BLI kinetics in C666-1 cells infected at a moiety of infection of 1 infectious unit/cell, measured at 12, 24, 48, 72, 96, and 120 hours after infection. (c) 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt cell viability assay was performed on the same cells infected with a moiety of infection of 1 infectious unit/cell, measured immediately after imaging. All data points for (a), (b) and (c) represent the mean ± SEM, with each experiment having been repeated three independent times and each experiment conducted in triplicates.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
In vivo longitudinal bioluminescence kinetics and biodistribution assays of C666-1 tumor-bearing mice injected intravenously with 3 × 108 infectious units of adv.oriP.luc, adv.SV40luc, or adv.oriP.E1A.oriP.luc.(a) Biodistribution after 6 hours (top panel) and 72 hours (bottom panel). (b) Tumor luciferase signals were quantified by drawing regions of interest around the imaged mouse tumors at 6, 24, 48, 72, 96, 120, 144, and 168 hours after infection. Each group comprised four mice. (c) Liver luciferase signals were quantified by drawing regions of interest around the liver at 6, 24, 48, 72, 96, 120, 144, and 168 hours after infection. Each data point represents the mean ± SD.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
In vivo evidence of adenoviral replication, specificity, and toxicity. (a) Changes in adenoviral DNA copy number in liver and tumor tissues 72 hours after infection with adv.oriP.E1A.oriP.luc or adv.oriP.luc, relative to number 6 hours after infection, determined by quantitative real-time PCR. An asterisk denotes statistical significance (P ≤ 0.05). (b) Immunohistochemistry for luciferase expression (bottom panel) and hematoxylin and eosin (H&E) staining (top panel) of livers infected with adv.oriP.E1A.oriP.luc after 6 and 72 hours. Black arrows indicate viral inclusion bodies, with ×3 magnification at the top right-hand corner. The scale-bar (in black) represents 100μm. (c) The effect of adenoviruses on tumor growth; a dagger denotes the death of a mouse. (d) The average ratio of liver-to-tumor signal for each mouse plotted for each time point imaged, as a measure of tumor specificity.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
The effect of the four interventions on the in vivobiodistribution of adv.oriP.luc. (a) Tumor signals for each intervention 24, 48 and 72 hours after intravenous infection with 3 × 108 infectious units of the virus in C666-1 tumor–bearing mice. (b) Liver signals from these same animals after 24, 48 and 72 hours. (c) Average tumor-to-liver ratios (TLRs) for each mouse indicate that STI571 is the most successful intervention in terms of modulating the biodistribution. (d) Changes in liver and tumor adenoviral DNA content, and the resulting TLR, in C666-1 tumor–bearing mice treated with either STI571 or vehicle (pH 4.2 H2O), 72 hours after infection; determined by quantitative real-time PCR. (e) Liver and tumor bioluminescence signals and the TLR in C17 tumor–bearing mice treated with STI571 or vehicle, 72 hours after infection. Asterisks denote statistical significance (P ≤ 0.05).
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

7. Figure 6
The effect of STI571 (50 mg/kg intraperitoneally × 4 days) on the tumor microenvironment. (a) STI571 significantly reduced interstitial fluid pressure in C666-1 tumor–bearing mice as measured by the wick-in-needle method compared with vehicle-treated mice (P = 0.05, N = 9 mice per group). Each data point represents mean ± SD. (b) Immunohistochemistry for phosphorylated platelet-derived growth factor-β receptor (PDGFβR) in a vehicle-treated (top) versus STI571-treated (bottom) tumor, demonstrating that STI571 inhibits phosphorylation of PDGFβR. Wide-field (left) and close-up (right) windows are shown. The scale bars (in black) on both windows represent 200 μm.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

8. Figure 7
Evaluation of kinetics, biodistribution, and therapeutic effect in C666-1 tumor–bearing mice injected with adv.oriP.E1A.oriP.luc(3 × 108 infectious units intra-tumorally (i.p.)) treated with STI571 or vehicle. Each group comprised five mice. (a) Tumor signals measured from mice imaged over time, where the final time point represents the average survival time. (b) Tumor growth over time of (i) vehicle alone (sterile water, pH 4.2, i.p. × 4 days), (ii) STI571 alone (50 mg/kg i.p. × 4 days), (iii) adv.oriP.luc plus vehicle, (iv) adv.oriP.E1A.oriP.luc plus vehicle, and (v) adv.oriP.E1A.oriP.luc plus STI571. Each data point represents the mean ± SD.
Molecular Therapy  , DOI: ( /mt.sj )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions