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Research ArticleBasic Science Investigation

Multimodality Imaging of Gene Transfer with a Receptor-Based Reporter Gene

Ron Chen, Jesse J. Parry, Walter J. Akers, Mikhail Y. Berezin, Issam M. El Naqa, Samuel Achilefu, W. Barry Edwards and Buck E. Rogers
Journal of Nuclear Medicine September 2010, 51 (9) 1456-1463; DOI: https://doi.org/10.2967/jnumed.109.063586
Ron Chen
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Jesse J. Parry
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Walter J. Akers
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Mikhail Y. Berezin
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Issam M. El Naqa
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Samuel Achilefu
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W. Barry Edwards
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Buck E. Rogers
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  • FIGURE 1.
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    FIGURE 1.

    In vitro evaluation of AdSSTR2, AdSSTR2-EGFP, and AdEGFP. (A) Representative saturation binding curves for 111In-DTPA-Y3-octreotate on membrane preparations from SCC-9 cells infected with AdSSTR2 or AdSSTR2-EGFP at 100 PFU per cell. Each data point represents mean ± SEM of triplicate measurements. (B) Fluorescence spectra of SCC-9 cells infected with AdEGFP or AdSSTR2-EGFP at 100 PFU per cell. norm = normalized.

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    FIGURE 2.

    Specific internalization of 111In-DTPA-Y3-octreotate at 37°C into SCC-9 cells infected with AdSSTR2 or AdSSTR2-EGFP at 100 PFU per cell. 111In-DTPA-Y3-octreotate (∼1.5 nM) was incubated with cells for various times in presence or absence of inhibitor. Cells were acid washed to remove surface-bound radioactivity and then harvested to determine internalized radioactivity. Data for each time point are presented as mean ± SEM of 3 experiments each performed in triplicate.

  • FIGURE 3.
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    FIGURE 3.

    Biodistribution of 111In-DTPA-Y3-octreotate in mice bearing SCC-9 tumor xenografts. Tumors were injected directly with adenovirus vectors, and 111In-DTPA-Y3-octreotate was injected via tail vein 2 d later. Mice were sacrificed 4 h (A), 24 h (B), and 48 h (C) later (n = 5 for each group). Data are presented as % ID/g ± SEM.

  • FIGURE 4.
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    FIGURE 4.

    Representative coronal nano-SPECT/CT images of SCC-9 tumor–bearing mice at 4, 24, and 48 h after injection of 111In-DTPA-Y3-octreotate. Axillary tumors were injected directly with Opti-MEM (control), AdSSTR2, or AdSSTR2-EGFP 2 d before administration of 111In-DTPA-Y3-octreotate. Coronal images show uptake of 111In-DTPA-Y3-octreotate in AdSSTR2- and AdSSTR2-EGFP-injected tumors but not control tumors (white arrows) and clearance through kidneys (yellow arrows).

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    FIGURE 5.

    Representative in vivo (A–C) and ex vivo (D–F) fluorescence imaging of mice bearing SCC-9 tumors injected directly with AdSSTR2 (A and D), AdSSTR2-EGFP (B and E), or AdEGFP (C and F) 2 d earlier. Tumor (arrow) was not visualized in vivo after injection of AdSSTR2 (A, negative control) but could be seen after injection of AdSSTR2-EGFP (B) or AdEGFP (C). In ex vivo studies, tumor had higher autofluorescence than liver and spleen (E), and fluorescence was observed in tumor injected with AdSSTR2-EGFP (D) or AdEGFP (F). au = arbitrary units.

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    TABLE 1.

    Ex Vivo Fluorescent Imaging of Tissues 48 Hours After Direct Injection of Adenovirus Vectors into SCC-9 Tumors

    TissueAdSSTR2 (n = 6)AdSSTR2-EGFP (n = 8)AdEGFP (n = 5)
    Tumor261* ± 69687 ± 161997* ± 306
    Liver83† ± 12560†,‡ ± 15286‡ ± 40
    Spleen9 ± 311 ± 34 ± 1
    • ↵* Values were significantly different at P ≤ 0.05.

    • ↵† Values were significantly different at P ≤ 0.05.

    • ↵‡ Values were significantly different at P ≤ 0.05.

    • Data are expressed as mean ± SEM for each tissue in arbitrary units.

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Journal of Nuclear Medicine: 51 (9)
Journal of Nuclear Medicine
Vol. 51, Issue 9
September 1, 2010
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Multimodality Imaging of Gene Transfer with a Receptor-Based Reporter Gene
Ron Chen, Jesse J. Parry, Walter J. Akers, Mikhail Y. Berezin, Issam M. El Naqa, Samuel Achilefu, W. Barry Edwards, Buck E. Rogers
Journal of Nuclear Medicine Sep 2010, 51 (9) 1456-1463; DOI: 10.2967/jnumed.109.063586

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Multimodality Imaging of Gene Transfer with a Receptor-Based Reporter Gene
Ron Chen, Jesse J. Parry, Walter J. Akers, Mikhail Y. Berezin, Issam M. El Naqa, Samuel Achilefu, W. Barry Edwards, Buck E. Rogers
Journal of Nuclear Medicine Sep 2010, 51 (9) 1456-1463; DOI: 10.2967/jnumed.109.063586
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