Advertisement

Nuclear Medicine and Molecular Imaging

, Volume 52, Issue 5, pp 359–367 | Cite as

Tc-99m and Fluorescence-Labeled Anti-Flt1 Peptide as a Multimodal Tumor Imaging Agent Targeting Vascular Endothelial Growth Factor-Receptor 1

  • Myoung Hyoun Kim
  • Seul-Gi Kim
  • Dae-Weung Kim
Original Article
  • 19 Downloads

Abstract

Purpose

We developed a Tc-99m and fluorescence-labeled peptide, Tc-99m TAMRA-GHEG-ECG-GNQWFI, to target tumor cells, and evaluated the diagnostic performance as a dual-modality imaging agent for tumor in a murine model.

Methods

TAMRA-GHEG-ECG-GNQWFI was synthesized using Fmoc solid-phase peptide synthesis. Radiolabeling of TAMRA-GHEG-ECG-GNQWFI with Tc-99m was done using ligand exchange via tartrate. Binding affinity and in vitro cellular uptake studies were performed. Gamma camera imaging, biodistribution, and ex vivo imaging studies were performed in murine models with U87MG tumors. Tumor tissue slides were prepared and analyzed with immunohistochemistry using confocal microscopy.

Results

After radiolabeling procedures with Tc-99m, Tc-99m TAMRA-GHEG-ECG-GNQWFI complexes were prepared in high yield (> 95%). The Kd of Tc-99m TAMRA-GHEG-ECG-GNQWFI determined by saturation binding was 29.5 ± 4.5 nM. Confocal microscopy images of U87MG cells incubated with TAMRA-GHEG-ECG-GNQWFI showed strong fluorescence in the cytoplasm. Gamma camera imaging revealed substantial uptake of Tc-99m TAMRA-GHEG-ECG-GNQWFI in tumors. Tumor uptake was effectively blocked by the co-injection of an excess concentration of GNQWFI. Specific uptake of Tc-99m TAMRA-GHEG-ECG-GNQWFI was assessed by biodistribution, ex vivo imaging, and immunohistochemistry stain studies.

Conclusions

In vivo and in vitro studies revealed substantial and specific uptake of Tc-99m TAMRA-GHEG-ECG-GNQWFI in tumor cells. Tc-99m TAMRA-GHEG-ECG-GNQWFI could be a good candidate dual-modality imaging agent for tumors.

Keywords

Vascular endothelial growth factor-receptor 1 Anti-Flt1 peptide Tc-99m TAMRA Multimodal imaging 

Notes

Compliance with Ethical Standards

Conflict of Interest

Dae-Weung Kim declares that this work was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (MSIP: Ministry of Science, ICT and Future Planning) (2017R1C1B2001886). Myoung Hyoun Kim and Seul-Gi Kim declare that they have no conflict of interest.

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

Informed Consent

The institutional review board of our institute approved this retrospective study, and the requirement to obtain informed consent was waived.

References

  1. 1.
    Leung DW, Cachianes G, Kuang WJ, Goeddel DV, Ferrara N. Vascular endothelial growth factor is a secreted angiogenic mitogen. Science. 1989;246:1306–9.CrossRefPubMedGoogle Scholar
  2. 2.
    Yoshino Y, Aoyagi M, Tamaki M, Duan L, Morimoto T, Ohno K. Activation of p38 MAPK and/or JNK contributes to increased levels of VEGF secretion in human malignant glioma cells. Int J Oncol. 2006;29:981–7.PubMedGoogle Scholar
  3. 3.
    Thomas KA. Vascular endothelial growth factor, a potent and selective angiogenic agent. J Biol Chem. 1996;271:603–6.CrossRefPubMedGoogle Scholar
  4. 4.
    Millauer B, Wizigmann-Voos S, Schnurch H, Martinez R, Moller NP, Risau W, et al. High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell. 1993;72:835–46.CrossRefPubMedGoogle Scholar
  5. 5.
    Goel HL, Mercurio AM. VEGF targets the tumour cell. Nat Rev Cancer. 2013;13:871–82.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Cai W, Chen K, Mohamedali KA, Cao Q, Gambhir SS, Rosenblum MG, et al. PET of vascular endothelial growth factor receptor expression. J Nucl Med. 2006;47:2048–56.PubMedGoogle Scholar
  7. 7.
    Bae DG, Kim TD, Li G, Yoon WH, Chae CB. Anti-flt1 peptide, a vascular endothelial growth factor receptor 1-specific hexapeptide, inhibits tumor growth and metastasis. Clin Cancer Res. 2005;11:2651–61.CrossRefPubMedGoogle Scholar
  8. 8.
    Yoo SA, Yoon HJ, Kim HS, Chae CB, De Falco S, Cho CS, et al. Role of placenta growth factor and its receptor flt-1 in rheumatoid inflammation: a link between angiogenesis and inflammation. Arthritis Rheum. 2009;60:345–54.CrossRefPubMedGoogle Scholar
  9. 9.
    Oh EJ, Choi JS, Kim H, Joo CK, Hahn SK. Anti-Flt1 peptide–hyaluronate conjugate for the treatment of retinal neovascularization and diabetic retinopathy. Biomaterials. 2011;32:3115–23.CrossRefPubMedGoogle Scholar
  10. 10.
    Kong JS, Yoo SA, Kang JH, Ko W, Jeon S, Chae CB, et al. Suppression of neovascularization and experimental arthritis by D-form of anti-flt-1 peptide conjugated with mini-PEG(™). Angiogenesis. 2011;14:431–42.CrossRefPubMedGoogle Scholar
  11. 11.
    Yang KS, Lim JH, Kim TW, Kim MY, Kim Y, Chung S, et al. Vascular endothelial growth factor-receptor 1 inhibition aggravates diabetic nephropathy through eNOS signaling pathway in db/db mice. PLoS One. 2014;9:e94540.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Cai W, Chen X. Multimodality molecular imaging of tumor angiogenesis. J Nucl Med. 2008;49(Suppl 2):113S–28S.CrossRefPubMedGoogle Scholar
  13. 13.
    Moore A, Medarova Z, Potthast A, Dai G. In vivo targeting of underglycosylated MUC-1 tumor antigen using a multimodal imaging probe. Cancer Res. 2004;64:1821–7.CrossRefPubMedGoogle Scholar
  14. 14.
    Miller SJ, Lee CM, Joshi BP, Gaustad A, Seibel EJ, Wang TD. Targeted detection of murine colonic dysplasia in vivo with flexible multispectral scanning fiber endoscopy. J Biomed Opt. 2012;17:021103.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Adusumilli PS, Stiles BM, Chan MK, Eisenberg DP, Yu Z, Stanziale SF, et al. Real-time diagnostic imaging of tumors and metastases by use of a replication-competent herpes vector to facilitate minimally invasive oncological surgery. FASEB J. 2006;20:726–8.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    de Barros AB, Tsourkas A, Saboury B, Cardoso VN, Alavi A. Emerging role of radiolabeled nanoparticles as an effective diagnostic technique. EJNMMI Res. 2012;2:39.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Chen IY, Wu JC. Cardiovascular molecular imaging: focus on clinical translation. Circulation. 2011;123:425–43.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Kim DW, Kim WH, Kim MH, Kim CG. Novel Tc-99m labeled ELR-containing 6-mer peptides for tumor imaging in epidermoid carcinoma xenografts model: a pilot study. Ann Nucl Med. 2013;27:892–7.CrossRefPubMedGoogle Scholar
  19. 19.
    Kim DW, Kim WH, Kim MH, Kim CG. Synthesis and evaluation of novel Tc-99m labeled NGR-containing hexapeptides as tumor imaging agents. J Label Compd Radiopharm. 2015;58:30–5.CrossRefGoogle Scholar
  20. 20.
    Kim DW, Kim WH, Kim MH, Kim CG. Synthesis and evaluation of Tc-99m-labeled RRL-containing peptide as a non-invasive tumor imaging agent in a mouse fibrosarcoma model. Ann Nucl Med. 2015;29:779–85.CrossRefPubMedGoogle Scholar
  21. 21.
    Wu C, Wei J, Gao K, Wang Y. Dibenzothiazoles as novel amyloid-imaging agents. Bioorg Med Chem. 2007;15:2789–96.CrossRefPubMedGoogle Scholar
  22. 22.
    Bouteiller C, Clave G, Bernardin A, Chipon B, Massonneau M, Renard PY, et al. Novel water-soluble near-infrared cyanine dyes: synthesis, spectral properties, and use in the preparation of internally quenched fluorescent probes. Bioconjug Chem. 2007;18:1303–17.CrossRefPubMedGoogle Scholar
  23. 23.
    Jabir NR, Tabrez S, Ashraf GM, Shakil S, Damanhouri GA, Kamal MA. Nanotechnology-based approaches in anticancer research. Int J Nanomedicine. 2012;7:4391–408.PubMedPubMedCentralGoogle Scholar
  24. 24.
    King R, Surfraz MB, Finucane C, Biagini SC, Blower PJ, Mather SJ. 99mTc-HYNIC-gastrin peptides: assisted coordination of 99mTc by amino acid side chains results in improved performance both in vitro and in vivo. J Nucl Med. 2009;50:591–8.CrossRefPubMedGoogle Scholar
  25. 25.
    Orbay H, Hong H, Zhang Y, Cai WPET. SPECT imaging of hindlimb ischemia: focusing on angiogenesis and blood flow. Angiogenesis. 2013;16:279–87.CrossRefPubMedGoogle Scholar

Copyright information

© Korean Society of Nuclear Medicine 2018

Authors and Affiliations

  • Myoung Hyoun Kim
    • 1
  • Seul-Gi Kim
    • 2
  • Dae-Weung Kim
    • 1
    • 2
  1. 1.Department of Nuclear Medicine and Institute of Wonkwang Medical ScienceWonkwang University School of MedicineIksanRepublic of Korea
  2. 2.Research Unit of Molecular Imaging Agent (RUMIA)Wonkwang University School of MedicineIksanRepublic of Korea

Personalised recommendations