Structure-inherent targeting of near-infrared fluorophores for parathyroid and thyroid gland imaging

Journal name:
Nature Medicine
Volume:
21,
Pages:
192–197
Year published:
DOI:
doi:10.1038/nm.3728
Received
Accepted
Published online

Abstract

The typical method for creating targeted contrast agents requires covalent conjugation of separate targeting and fluorophore domains. In this study, we demonstrate that it is possible to create near-infrared (NIR) fluorophores with different tissue specificities driven by their inherent chemical structures. Thus, a single compact molecule performs both targeting and imaging. We use this strategy to solve a major problem in head and neck surgery: the identification and preservation of parathyroid and thyroid glands. We synthesized 700-nm and 800-nm halogenated fluorophores that show high uptake into these glands after a single intravenous (IV) injection of 0.06 mg kg−1 in a pig. By using a dual-channel NIR imaging system, we observed—in real time and with high sensitivity—the unambiguous distinction of parathyroid and thyroid glands simultaneously in the context of blood and surrounding soft tissue. This novel technology lays a foundation for performing head and neck surgery with increased precision and efficiency along with potentially lower morbidity, and it provides a general strategy for developing targeted NIR fluorophores.

At a glance

Figures

  1. Synthetic schemes for T700 and T800 NIR fluorophores.
    Figure 1: Synthetic schemes for T700 and T800 NIR fluorophores.
  2. Thyroid targeting efficiency in mice.
    Figure 2: Thyroid targeting efficiency in mice.

    (a,b) In vivo thyroid targeting of T700 (a) and T800 (b) NIR fluorophores in mice. Each fluorophore was IV injected into 20-g CD-1 mice (10 nmol; 0.2 mg kg−1) 1 h and 4 h before imaging. All NIR fluorescence images have identical exposure and normalizations. SG, salivary glands; TG, thyroid glands (arrows). Scale bars, 1 cm. Images are representative of n = 3 independent experiments at 1 h after injection. Error bars show means ± sd (n = 5 mice).

  3. In vivo parathyroid and thyroid imaging using T700-F and T800-F in pigs.
    Figure 3: In vivo parathyroid and thyroid imaging using T700-F and T800-F in pigs.

    Top, 5 μmol (0.06 mg kg−1) of T700-F and T800-F were injected IV into 35-kg Yorkshire pigs 5 h before imaging. All NIR fluorescence images have identical exposure and normalizations. Parathyroid signal to thyroid signal ratio (PTR) curves were determined from individual SBR curves. (ac) Shown are magnified images of parathyroid (a), thyroid and parathyroid glands 1 h (b) and 4 h (c) after injection. La, larynx; PG, parathyroid glands (arrowheads); TG, thyroid gland (arrows), Th, thymus. Scale bars, 1 cm. Each data point is the mean ± sd from n = 3 animals.

  4. Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in rats.
    Figure 4: Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in rats.

    (a) Dual-channel in vivo fluorescence imaging using both T700-F and T800-F in a rat. 0.2 μmol (0.35 mg kg−1) of T800-F was IV injected into a 250 g SD rat 24 h before imaging, followed by 0.2 μmol of T700-F injected 6 h before imaging. Es, esophagus; PG, parathyroid glands (arrowheads); SG, salivary glands; TG, thyroid glands (arrows); Tr, trachea. Scale bars, 1 cm. (b) H&E and NIR imaging of resected parathyroid and thyroid tissues from (a). Scale bars, 300 μm. All NIR fluorescence images for each condition have identical exposure times and normalizations. Pseudocolored red and green colors were used for 700-nm and 800-nm channel images, respectively, in the color-NIR merged image. Yellow in the merged image indicates colocalization of the T700 and T800 fluorophores.

  5. Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in pigs.
    Figure 5: Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in pigs.

    (a) Dual-channel in vivo fluorescence imaging using both T700-F and T800-F in a pig. 5 μmol (0.06 mg kg−1) of T800-F was IV injected into a 35 kg Yorkshire pig 4 h before dual imaging, followed by 5 μmol of T700-F injected 2 h later. La, larynx; PG, parathyroid glands (arrowheads); TG, thyroid gland (arrows). Scale bars, 1 cm. Pseudocolored red and green colors were used for 700-nm and 800-nm channel images, respectively, in the color-NIR merged image. (b) H&E and NIR imaging of resected parathyroid and thyroid tissues from a. Scale bars, 300 μm. All NIR fluorescence images have identical exposure and normalizations.

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Author information

  1. These authors contributed equally to this work.

    • Hoon Hyun &
    • Min Ho Park

Affiliations

  1. Department of Medicine, Division of Hematology and Oncology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA.

    • Hoon Hyun,
    • Min Ho Park,
    • Hideyuki Wada,
    • John V Frangioni &
    • Hak Soo Choi
  2. Department of Radiology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA.

    • John V Frangioni
  3. Department of Surgery, Chonnam National University Medical School, Gwangju, South Korea.

    • Min Ho Park
  4. Department of Chemistry, Georgia State University, Atlanta, Georgia, USA.

    • Eric A Owens &
    • Maged Henary
  5. Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, Georgia, USA.

    • Eric A Owens &
    • Maged Henary
  6. Department of Surgery, Leiden University Medical Center, Leiden, the Netherlands.

    • Henricus J M Handgraaf &
    • Alexander L Vahrmeijer
  7. Curadel, LLC, Worcester, Massachusetts, USA.

    • John V Frangioni
  8. Department of Cogno-Mechatronics Engineering, Pusan National University, Busan, South Korea.

    • Hak Soo Choi

Contributions

H.H., M.H.P., H.W., E.A.O. and H.J.M.H. performed the experiments. H.H., M.H.P., M.H., A.L.V., J.V.F. and H.S.C. reviewed, analyzed and interpreted the data. H.H., J.V.F. and H.S.C. wrote the paper. All authors discussed the results and commented on the manuscript.

Competing financial interests

J.V.F. is currently CEO of Curadel, LLC, which has licensed FLARE imaging systems and contrast agents from the Beth Israel Deaconess Medical Center.

Corresponding author

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Supplementary information

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  1. Supplementary Text and Figures (1,642 KB)

    Supplementary Methods and Supplementary Figures 1–5

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