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OtherBasic Science Investigations

64Cu-Labeled Folate-Conjugated Shell Cross-Linked Nanoparticles for Tumor Imaging and Radiotherapy: Synthesis, Radiolabeling, and Biologic Evaluation

Raffaella Rossin, Dipanjan Pan, Kai Qi, Jeffrey L. Turner, Xiankai Sun, Karen L. Wooley and Michael J. Welch
Journal of Nuclear Medicine July 2005, 46 (7) 1210-1218;
Raffaella Rossin
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Dipanjan Pan
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Kai Qi
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Jeffrey L. Turner
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Xiankai Sun
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Karen L. Wooley
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Michael J. Welch
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  • FIGURE 1.
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    FIGURE 1.

    Preparation of SCKs functionalized with folate for targeting and labeled with FTSC or TETA for fluorescence or radionuclide detection, respectively. Reagents and conditions: (i) polymer micelle formation: tetrahydrofuran, followed by controlled addition of water and dialysis against water; (ii) shell crosslinking: block copolymer micelles, 2,2′-(ethylenedioxy)-bis(ethylamine), 1-(3′-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) methiodide, room temperature (RT), followed by dialysis against water; (iii) shell functionalization with folate-PEG1,600-amine (PEG1,600 is a poly(ethylene glycol) spacer with a molecular weight of 1,600 Da): nonfunctionalized SCKs, folate-PEG1,600-amine, EDC methiodide, RT, followed by dialysis against sodium phosphate-buffered saline at pH 7.3; (iv) shell functionalization with fluorescein thiosemicarbazide (FTSC): SCK-folate, FTSC, EDC methiodide, RT, followed by dialysis against sodium phosphate-buffered saline at pH 7.3; (v) shell functionalization with TETA-amine: SCK-folate, sulfo-NHS (s-NHS), EDC, 4°C, 2 h, followed by Centricon separation, TETA-amine, 4°C, overnight.

  • FIGURE 2.
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    FIGURE 2.

    Fluorescence micrographs of KB cells incubated for 4 h at 37°C (A and B) and 4°C (C and D) with FTSC and folate-conjugated SCKs (10 μg FTSC-SCK-folate per 33-mm culture dish, plated with 3 × 105 cells per dish 24 h before assay). Incubation at 4°C led to surface uptake of FTSC-SCK-folate onto KB cells (C), whereas fluorescent endocytotic vesicles are visible inside the cells after 4 h of incubation at 37°C (A). At both incubation temperatures, excess folic acid (1 mmol/L) competitively inhibited FTSC-SCK-folate cell uptake (B and D).

  • FIGURE 3.
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    FIGURE 3.

    Dependence of tumor uptake on tumor size 4 h after injection of 64Cu-labeled folated (A) and nonfolated (B) SCKs in KB xenograft-bearing mice. Data are presented as %ID/g. Within each experimental group, small tumors exhibited enhanced nanoparticle uptake. Coadministration of excess folic acid led to competitive block of 64Cu-TETA-SCK-folate tumor uptake (A, dashed line), whereas it had no noticeable effect on 64Cu-TETA-SCK tumor uptake (B, dashed line).

  • FIGURE 4.
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    FIGURE 4.

    Dual-tracer autoradiographs demonstrate regional concentration of 64Cu-labeled folated (A) or nonfolated (B) SCKs in mice bearing 0.3- to 0.6-g KB xenografts. Tumors were resected and sectioned into 1-mm slices 1 h after 18F-FDG administration and 24 h after administration of 64Cu-radiolabeled SCKs. Images of 18F-FDG distribution were collected shortly after tumor resection, whereas distribution of 64Cu-labeled SCKs in the same slices was imaged with a 20-h delay. Both tumors exhibit scattered necrotic regions (no radiotracer uptake).

  • FIGURE 5.
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    FIGURE 5.

    Cross-sectional slice from a tumor generated by subcutaneous inoculation of a nu/nu athymic mouse with 1 × 106 human KB cells. Tumor mass is enclosed in a capsule (C) and it is characterized by scattered necrotic regions (N) surrounded by viable tissues (V). (hematoxylin–eosin, ×40)

Tables

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    TABLE 1

    Biodistribution Data of 64Cu-TETA-SCK-Folate in KB Tumor Cell Xenograft-Bearing Athymic Nude Female Mice (n = 4)

    Biodistribution%ID/g%ID/organ
    10 min1 h4 h24 h10 min1 h4 h24 h
    Blood2.1 ± 0.22.1 ± 0.22.5 ± 0.32.6 ± 0.63.1 ± 0.23.4 ± 0.24.2 ± 0.34.3 ± 0.9
    Lung39.7 ± 5.110.9 ± 3.814.4 ± 5.910.8 ± 1.45.1 ± 0.91.5 ± 0.22.1 ± 0.91.5 ± 0.2
    Liver56.0 ± 7.138.4 ± 7.933.8 ± 2.621.5 ± 2.952.3 ± 5.641.7 ± 3.034.1 ± 2.525.2 ± 2.9
    Spleen18.3 ± 6.310.8 ± 3.28.8 ± 0.54.7 ± 1.01.5 ± 0.41.0 ± 0.30.8 ± 0.20.5 ± 0.2
    Kidney7.2 ± 1.68.1 ± 1.87.9 ± 0.88.5 ± 1.31.2 ± 0.31.3 ± 0.21.4 ± 0.21.5 ± 0.4
    Muscle0.8 ± 0.10.8 ± 0.20.9 ± 0.21.0 ± 0.17.2 ± 0.57.5 ± 1.38.6 ± 1.79.5 ± 1.3
    Heart2.3 ± 0.22.7 ± 0.53.7 ± 0.34.3 ± 0.50.2 ± 0.00.3 ± 0.10.4 ± 0.00.5 ± 0.0
    Tumor3.4 ± 1.22.3 ± 0.75.9 ± 2.82.9 ± 3.10.04 ± 0.020.12 ± 0.020.04 ± 0.020.09 ± 0.07
    • Data are presented as percentage injected dose per gram (%ID/g) and percentage injected dose per organ (%ID/organ) ± SD.

    • View popup
    TABLE 2

    Biodistribution Data of 64Cu-TETA-SCK in KB Tumor Cell Xenograft-Bearing Athymic Nude Female Mice (n = 4)

    Biodistribution%ID/g%ID/organ
    10 min1 h4 h24 h10 min1 h4 h24 h
    Blood3.3 ± 0.32.4 ± 0.72.6 ± 0.83.1 ± 0.35.1 ± 0.43.7 ± 0.83.8 ± 1.34.8 ± 0.2
    Lung32.5 ± 14.015.0 ± 4.113.8 ± 6.612.7 ± 1.84.6 ± 1.81.9 ± 0.31.8 ± 0.91.8 ± 0.1
    Liver45.7 ± 3.535.3 ± 8.831.8 ± 11.926.9 ± 3.945.6 ± 2.534.3 ± 7.327.3 ± 10.226.1 ± 1.0
    Spleen11.5 ± 2.58.2 ± 3.44.7 ± 2.04.5 ± 0.61.0 ± 0.20.8 ± 0.30.4 ± 0.20.4 ± 0.1
    Kidney9.0 ± 0.68.6 ± 2.09.5 ± 0.810.8 ± 1.11.5 ± 0.01.4 ± 0.41.4 ± 0.31.7 ± 0.1
    Muscle1.1 ± 0.10.9 ± 0.20.9 ± 0.21.4 ± 0.210.1 ± 0.28.4 ± 2.07.6 ± 1.913.1 ± 2.6
    Heart3.7 ± 0.14.0 ± 1.34.7 ± 1.56.2 ± 0.40.4 ± 0.00.5 ± 0.20.4 ± 0.20.6 ± 0.0
    Tumor2.2 ± 0.33.2 ± 0.76.0 ± 1.95.6 ± 0.90.04 ± 0.040.17 ± 0.070.13 ± 0.040.18 ± 0.11
    • Data are presented as percentage injected dose per gram (%ID/g) and percentage injected dose per organ (%ID/organ) ± SD.

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Journal of Nuclear Medicine: 46 (7)
Journal of Nuclear Medicine
Vol. 46, Issue 7
July 1, 2005
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64Cu-Labeled Folate-Conjugated Shell Cross-Linked Nanoparticles for Tumor Imaging and Radiotherapy: Synthesis, Radiolabeling, and Biologic Evaluation
Raffaella Rossin, Dipanjan Pan, Kai Qi, Jeffrey L. Turner, Xiankai Sun, Karen L. Wooley, Michael J. Welch
Journal of Nuclear Medicine Jul 2005, 46 (7) 1210-1218;

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64Cu-Labeled Folate-Conjugated Shell Cross-Linked Nanoparticles for Tumor Imaging and Radiotherapy: Synthesis, Radiolabeling, and Biologic Evaluation
Raffaella Rossin, Dipanjan Pan, Kai Qi, Jeffrey L. Turner, Xiankai Sun, Karen L. Wooley, Michael J. Welch
Journal of Nuclear Medicine Jul 2005, 46 (7) 1210-1218;
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