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

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.

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Figure 1
Figure 2: Thyroid targeting efficiency in mice.
Figure 3: In vivo parathyroid and thyroid imaging using T700-F and T800-F in pigs.
Figure 4: Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in rats.
Figure 5: Simultaneous in vivo NIR imaging of parathyroid and thyroid glands in pigs.

References

  1. 1

    Lee, J.H., Park, G., Hong, G.H., Choi, J. & Choi, H.S. Design considerations for targeted optical contrast agents. Quant. Imaging Med. Surg. 2, 266–273 (2012).

    Google Scholar 

  2. 2

    Kobayashi, H., Ogawa, M., Alford, R., Choyke, P.L. & Urano, Y. New strategies for fluorescent probe design in medical diagnostic imaging. Chem. Rev. 110, 2620–2640 (2010).

    CAS  Article  Google Scholar 

  3. 3

    Achilefu, S. The insatiable quest for near-infrared fluorescent probes for molecular imaging. Angew. Chem. Int. Edn Engl. 49, 9816–9818 (2010).

    CAS  Article  Google Scholar 

  4. 4

    Pogue, B.W., Leblond, F., Krishnaswamy, V. & Paulsen, K.D. Radiologic and near-infrared/optical spectroscopic imaging: where is the synergy? AJR Am. J. Roentgenol. 195, 321–332 (2010).

    Article  Google Scholar 

  5. 5

    Gao, J. et al. Ultrasmall near-infrared non-cadmium quantum dots for in vivo tumor imaging. Small 6, 256–261 (2010).

    CAS  Article  Google Scholar 

  6. 6

    Vahrmeijer, A.L., Hutteman, M., van der Vorst, J.R., van de Velde, C.J. & Frangioni, J.V. Image-guided cancer surgery using near-infrared fluorescence. Nat. Rev. Clin. Oncol. 10, 507–518 (2013).

    CAS  Article  Google Scholar 

  7. 7

    Wang, T.S. Endocrine surgery. Am. J. Surg. 202, 369–371 (2011).

    Article  Google Scholar 

  8. 8

    Lin, D.T., Patel, S.G., Shaha, A.R., Singh, B. & Shah, J.P. Incidence of inadvertent parathyroid removal during thyroidectomy. Laryngoscope 112, 608–611 (2002).

    Article  Google Scholar 

  9. 9

    Frilling, A. & Weber, F. Complications in Thyroid and Parathyroid Surgery (Springer, 2007).

  10. 10

    Fancy, T., Gallagher, D. III & Hornig, J.D. Surgical anatomy of the thyroid and parathyroid glands. Otolaryngol. Clin. North Am. 43, 221–227, vii (2010).

    Article  Google Scholar 

  11. 11

    Bliss, R.D., Gauger, P.G. & Delbridge, L.W. Surgeon's approach to the thyroid gland: surgical anatomy and the importance of technique. World J. Surg. 24, 891–897 (2000).

    CAS  Article  Google Scholar 

  12. 12

    van der Vorst, J.R. et al. Intraoperative near-infrared fluorescence imaging of parathyroid adenomas using low-dose methylene blue. Head Neck 36, 853–858 (2014).

    Article  Google Scholar 

  13. 13

    Gioux, S., Choi, H.S. & Frangioni, J.V. Image-guided surgery using invisible near-infrared light: fundamentals of clinical translation. Mol. Imaging 9, 237–255 (2010).

    CAS  Article  Google Scholar 

  14. 14

    Mieog, J.S. et al. Toward optimization of imaging system and lymphatic tracer for near-infrared fluorescent sentinel lymph node mapping in breast cancer. Ann. Surg. Oncol. 18, 2483–2491 (2011).

    Article  Google Scholar 

  15. 15

    Soshin, T. et al. A method for sampling and tissue preparation of the parathyroid glands in miniature pigs for toxicity studies. J. Toxicol. Sci. 35, 235–238 (2010).

    CAS  Article  Google Scholar 

  16. 16

    Kittel, B. et al. Revised guides for organ sampling and trimming in rats and mice-Part 2. Exp. Toxicol. Pathol. 55, 413–431 (2004).

    Article  Google Scholar 

  17. 17

    Choi, H.S. et al. Targeted zwitterionic near-infrared fluorophores for improved optical imaging. Nat. Biotechnol. 31, 148–153 (2013).

    CAS  Article  Google Scholar 

  18. 18

    Gibbs-Strauss, S.L. et al. Nerve-highlighting fluorescent contrast agents for image-guided surgery. Mol. Imaging 10, 91–101 (2011).

    CAS  Article  Google Scholar 

  19. 19

    Gibbs, S.L. et al. Structure-activity relationship of nerve-highlighting fluorophores. PLoS ONE 8, e73493 (2013).

    CAS  Article  Google Scholar 

  20. 20

    Pollack, G., Pollack, A., Delfiner, J. & Fernandez, J. Parathyroid surgery and methylene blue: A review with guidelines for safe intraoperative use. Laryngoscope 119, 1941–1946 (2009).

    Article  Google Scholar 

  21. 21

    McWade, M.A. et al. A novel optical approach to intraoperative detection of parathyroid glands. Surgery 154, 1371–1377, discussion 1377 (2013).

    Article  Google Scholar 

  22. 22

    Paras, C., Keller, M., White, L., Phay, J. & Mahadevan-Jansen, A. Near-infrared autofluorescence for the detection of parathyroid glands. J. Biomed. Opt. 16, 067012 (2011).

    Article  Google Scholar 

  23. 23

    Lim, Y.T. et al. Selection of quantum dot wavelengths for biomedical assays and imaging. Mol. Imaging 2, 50–64 (2003).

    CAS  Article  Google Scholar 

  24. 24

    Sens, R. & Drexhage, K.H. Fluorescence quantum yield of oxazine and carbazine laser dyes. J. Lumin. 24–25, 709–712 (1981).

    Article  Google Scholar 

  25. 25

    Benson, C. & Kues, H.A. Absorption and fluorescence properties of cyanine dyes. J. Chem. Eng. Data 22, 379–383 (1977).

    CAS  Article  Google Scholar 

  26. 26

    Troyan, S.L. et al. The FLARE intraoperative near-infrared fluorescence imaging system: a first-in-human clinical trial in breast cancer sentinel lymph node mapping. Ann. Surg. Oncol. 16, 2943–2952 (2009).

    Article  Google Scholar 

  27. 27

    Ashitate, Y. et al. Two-wavelength near-infrared fluorescence for the quantitation of drug antiplatelet effects in large animal model systems. J. Vasc. Surg. 56, 171–180 (2012).

    Article  Google Scholar 

  28. 28

    Choi, H.S. et al. Rapid translocation of nanoparticles from the lung airspaces to the body. Nat. Biotechnol. 28, 1300–1303 (2010).

    CAS  Article  Google Scholar 

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Acknowledgements

We thank D. Burrington, Jr. for editing and E. Trabucchi for administrative assistance. This study was supported by the following grants from the National Institutes of Health: NCI BRP grant no. R01-CA-115296 (J.V.F.), NIBIB grant nos. R01-EB-010022 (J.V.F.) and R01-EB-011523 (H.S.C.), and a grant from the Dana Foundation in brain and immuno-imaging (H.S.C.). The contents of this paper are solely the responsibility of the authors and do not necessarily reflect the official views of the National Institutes of Health

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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.

Corresponding author

Correspondence to Hak Soo Choi.

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Competing 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.

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Supplementary Methods and Supplementary Figures 1–5 (PDF 1603 kb)

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Hyun, H., Park, M., Owens, E. et al. Structure-inherent targeting of near-infrared fluorophores for parathyroid and thyroid gland imaging. Nat Med 21, 192–197 (2015). https://doi.org/10.1038/nm.3728

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