Skip to main content
SpringerLink
Log in

Evaluation of two novel 64Cu-labeled RGD peptide radiotracers for enhanced PET imaging of tumor integrin αvβ3

  • Original Article
  • Published:
European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Our goal was to demonstrate that suitably derivatized monomeric RGD peptide-based PET tracers, targeting integrin αvβ3, may offer advantages in image contrast, time for imaging, and low uptake in nontarget tissues.

Methods

Two cyclic RGDfK derivatives, (PEG)2-c(RGDfK) and PEG4-SAA4-c(RGDfK), were constructed and conjugated to NOTA for 64Cu labeling. Their integrin αvβ3-binding properties were determined via a competitive cell binding assay. Mice bearing U87MG tumors were intravenously injected with each of the 64Cu-labeled peptides, and PET scans were acquired during the first 30 min, and 2 and 4 h after injection. Blocking and ex vivo biodistribution studies were carried out to validate the PET data and confirm the specificity of the tracers.

Results

The IC50 values of NOTA-(PEG)2-c(RGDfK) and NOTA-PEG4-SAA4-c(RGDfK) were 444 ± 41 nM and 288 ± 66 nM, respectively. Dynamic PET data of 64Cu-NOTA-(PEG)2-c(RGDfK) and 64Cu-NOTA-PEG4-SAA4-c(RGDfK) showed similar circulation t 1/2 and peak tumor uptake of about 4 %ID/g for both tracers. Due to its marked hydrophilicity, 64Cu-NOTA-PEG4-SAA4-c(RGDfK) provided faster clearance from tumor and normal tissues yet maintained excellent tumor-to-background ratios. Static PET scans at later time-points corroborated the enhanced excretion of the tracer, especially from abdominal organs. Ex vivo biodistribution and receptor blocking studies confirmed the accuracy of the PET data and the integrin αvβ3-specificity of the peptides.

Conclusion

Our two novel RGD-based radiotracers with optimized pharmacokinetic properties allowed fast, high-contrast PET imaging of tumor-associated integrin αvβ3. These tracers may facilitate the imaging of abdominal malignancies, normally precluded by high background uptake.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Hwang R, Varner J. The role of integrins in tumor angiogenesis. Hematol Oncol Clin North Am. 2004;18:991–1006.

    Article  PubMed  Google Scholar 

  2. Robinson SD, Hodivala-Dilke KM. The role of beta3-integrins in tumor angiogenesis: context is everything. Curr Opin Cell Biol. 2011;23:630–7.

    Article  CAS  PubMed  Google Scholar 

  3. Puduvalli VK. Inhibition of angiogenesis as a therapeutic strategy against brain tumors. Cancer Treat Res. 2004;117:307–36.

    Article  CAS  PubMed  Google Scholar 

  4. Sengupta S, Chattopadhyay N, Mitra A, Ray S, Dasgupta S, Chatterjee A. Role of alphavbeta3 integrin receptors in breast tumor. J Exp Clin Cancer Res. 2001;20:585–90.

    CAS  PubMed  Google Scholar 

  5. Felding-Habermann B, Fransvea E, O'Toole TE, Manzuk L, Faha B, Hensler M. Involvement of tumor cell integrin alpha v beta 3 in hematogenous metastasis of human melanoma cells. Clin Exp Metastasis. 2002;19:427–36.

    Article  CAS  PubMed  Google Scholar 

  6. Jin H, Varner J. Integrins: roles in cancer development and as treatment targets. Br J Cancer. 2004;90:561–5.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Kumar CC. Integrin alpha v beta 3 as a therapeutic target for blocking tumor-induced angiogenesis. Curr Drug Targets. 2003;4:123–31.

    Article  CAS  PubMed  Google Scholar 

  8. Kumar CC, Armstrong L, Yin Z, Malkowski M, Maxwell E, Ling H, et al. Targeting integrins alpha v beta 3 and alpha v beta 5 for blocking tumor-induced angiogenesis. Adv Exp Med Biol. 2000;476:169–80.

    Article  CAS  PubMed  Google Scholar 

  9. Cai W, Niu G, Chen X. Imaging of integrins as biomarkers for tumor angiogenesis. Curr Pharm Des. 2008;14:2943–73.

    Article  CAS  PubMed  Google Scholar 

  10. Schottelius M, Laufer B, Kessler H, Wester HJ. Ligands for mapping alphavbeta3-integrin expression in vivo. Acc Chem Res. 2009;42:969–80.

    Article  CAS  PubMed  Google Scholar 

  11. Gaertner FC, Kessler H, Wester HJ, Schwaiger M, Beer AJ. Radiolabelled RGD peptides for imaging and therapy. Eur J Nucl Med Mol Imaging. 2012;39 Suppl 1:S126–38.

    Article  PubMed  Google Scholar 

  12. Kenny LM, Coombes RC, Oulie I, Contractor KB, Miller M, Spinks TJ, et al. Phase I trial of the positron-emitting Arg-Gly-Asp (RGD) peptide radioligand 18F-AH111585 in breast cancer patients. J Nucl Med. 2008;49:879–86.

    Article  PubMed  Google Scholar 

  13. Beer AJ, Haubner R, Sarbia M, Goebel M, Luderschmidt S, Grosu AL, et al. Positron emission tomography using [18F]Galacto-RGD identifies the level of integrin alpha(v)beta3 expression in man. Clin Cancer Res. 2006;12:3942–9.

    Article  CAS  PubMed  Google Scholar 

  14. Jeong JM, Hong MK, Chang YS, Lee YS, Kim YJ, Cheon GJ, et al. Preparation of a promising angiogenesis PET imaging agent: 68Ga-labeled c(RGDyK)-isothiocyanatobenzyl-1,4,7-triazacyclononane-1,4,7-triacetic acid and feasibility studies in mice. J Nucl Med. 2008;49:830–6.

    Article  CAS  PubMed  Google Scholar 

  15. Hernandez R, Valdovinos HF, Yang Y, Chakravarty R, Hong H, Barnhart TE, et al. (44)Sc: an attractive isotope for peptide-based PET imaging. Mol Pharm. 2014;11:2954–61.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Kim SJ, Lee JS, Im KC, Kim SY, Park SA, Lee SJ, et al. Kinetic modeling of 3'-deoxy-3'-18F-fluorothymidine for quantitative cell proliferation imaging in subcutaneous tumor models in mice. J Nucl Med. 2008;49:2057–66.

    Article  PubMed  Google Scholar 

  17. Haubner R, Decristoforo C. Radiolabelled RGD peptides and peptidomimetics for tumour targeting. Front Biosci (Landmark Ed). 2009;14:872–86.

    Article  CAS  Google Scholar 

  18. Bogdanowich-Knipp SJ, Chakrabarti S, Williams TD, Dillman RK, Siahaan TJ. Solution stability of linear vs. cyclic RGD peptides. J Pept Res. 1999;53:530–41.

    Article  CAS  PubMed  Google Scholar 

  19. Samanen J, Ali F, Romoff T, Calvo R, Sorenson E, Vasko J, et al. Development of a small RGD peptide fibrinogen receptor antagonist with potent antiaggregatory activity in vitro. J Med Chem. 1991;34:3114–25.

    Article  CAS  PubMed  Google Scholar 

  20. Haubner R, Wester HJ, Burkhart F, Senekowitsch-Schmidtke R, Weber W, Goodman SL, et al. Glycosylated RGD-containing peptides: tracer for tumor targeting and angiogenesis imaging with improved biokinetics. J Nucl Med. 2001;42:326–36.

    CAS  PubMed  Google Scholar 

  21. Haubner R, Wester HJ, Weber WA, Mang C, Ziegler SI, Goodman SL, et al. Noninvasive imaging of alpha(v)beta3 integrin expression using 18F-labeled RGD-containing glycopeptide and positron emission tomography. Cancer Res. 2001;61:1781–5.

    CAS  PubMed  Google Scholar 

  22. Li ZB, Cai W, Cao Q, Chen K, Wu Z, He L, et al. (64)Cu-labeled tetrameric and octameric RGD peptides for small-animal PET of tumor alpha(v)beta(3) integrin expression. J Nucl Med. 2007;48:1162–71.

    Article  CAS  PubMed  Google Scholar 

  23. Liu S. Radiolabeled multimeric cyclic RGD peptides as integrin alphavbeta3 targeted radiotracers for tumor imaging. Mol Pharm. 2006;3:472–87.

    Article  CAS  PubMed  Google Scholar 

  24. Auzzas L, Zanardi F, Battistini L, Burreddu P, Carta P, Rassu G, et al. Targeting alphavbeta3 integrin: design and applications of mono- and multifunctional RGD-based peptides and semipeptides. Curr Med Chem. 2010;17:1255–99.

    Article  CAS  PubMed  Google Scholar 

  25. Dijkgraaf I, Yim CB, Franssen GM, Schuit RC, Luurtsema G, Liu S, et al. PET imaging of alphavbeta(3) integrin expression in tumours with 68Ga-labelled mono-, di- and tetrameric RGD peptides. Eur J Nucl Med Mol Imaging. 2011;38:128–37.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Notni J, Pohle K, Wester HJ. Be spoilt for choice with radiolabelled RGD peptides: preclinical evaluation of 68Ga-TRAP(RGD)(3). Nucl Med Biol. 2013;40:33–41.

    Article  CAS  PubMed  Google Scholar 

  27. Knetsch PA, Zhai C, Rangger C, Blatzer M, Haas H, Kaeopookum P, et al. [(68)Ga]FSC-(RGD)3 a trimeric RGD peptide for imaging alphavbeta3 integrin expression based on a novel siderophore derived chelating scaffold-synthesis and evaluation. Nucl Med Biol. 2015;42:115–22.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Shi J, Kim YS, Zhai S, Liu Z, Chen X, Liu S. Improving tumor uptake and pharmacokinetics of (64)Cu-labeled cyclic RGD peptide dimers with Gly(3) and PEG(4) linkers. Bioconjug Chem. 2009;20:750–9.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Ji S, Czerwinski A, Zhou Y, Shao G, Valenzuela F, Sowinski P, et al. (99m)Tc-Galacto-RGD2: a novel 99mTc-labeled cyclic RGD peptide dimer useful for tumor imaging. Mol Pharm. 2013;10:3304–14.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Liu S, Liu Z, Chen K, Yan Y, Watzlowik P, Wester HJ, et al. 18F-labeled galacto and PEGylated RGD dimers for PET imaging of alphavbeta3 integrin expression. Mol Imaging Biol. 2010;12:530–8.

    Article  PubMed Central  PubMed  Google Scholar 

  31. Jung KH, Lee KH, Paik JY, Ko BH, Bae JS, Lee BC, et al. Favorable biokinetic and tumor-targeting properties of 99mTc-labeled glucosamino RGD and effect of paclitaxel therapy. J Nucl Med. 2006;47:2000–7.

    CAS  PubMed  Google Scholar 

  32. Dumont RA, Deininger F, Haubner R, Maecke HR, Weber WA, Fani M. Novel (64)Cu- and (68)Ga-labeled RGD conjugates show improved PET imaging of alpha(nu)beta(3) integrin expression and facile radiosynthesis. J Nucl Med. 2011;52:1276–84.

    Article  CAS  PubMed  Google Scholar 

  33. Zhang X, Xiong Z, Wu Y, Cai W, Tseng JR, Gambhir SS, et al. Quantitative PET imaging of tumor integrin alphavbeta3 expression with 18F-FRGD2. J Nucl Med. 2006;47:113–21.

    PubMed Central  CAS  PubMed  Google Scholar 

  34. Wu Z, Li ZB, Chen K, Cai W, He L, Chin FT, et al. microPET of tumor integrin alphavbeta3 expression using 18F-labeled PEGylated tetrameric RGD peptide (18F-FPRGD4). J Nucl Med. 2007;48:1536–44.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Behr TM, Goldenberg DM, Becker W. Reducing the renal uptake of radiolabeled antibody fragments and peptides for diagnosis and therapy: present status, future prospects and limitations. Eur J Nucl Med. 1998;25:201–12.

    Article  CAS  PubMed  Google Scholar 

  36. Cai W, Chen X. Multimodality molecular imaging of tumor angiogenesis. J Nucl Med. 2008;49 Suppl 2:113S–28.

  37. Doss M, Kolb HC, Zhang JJ, Belanger MJ, Stubbs JB, Stabin MG, et al. Biodistribution and radiation dosimetry of the integrin marker 18F-RGD-K5 determined from whole-body PET/CT in monkeys and humans. J Nucl Med. 2012;53:787–95.

    Article  PubMed  Google Scholar 

  38. Wan W, Guo N, Pan D, Yu C, Weng Y, Luo S, et al. First experience of 18F-alfatide in lung cancer patients using a new lyophilized kit for rapid radiofluorination. J Nucl Med. 2013;54:691–8.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  39. Liu Z, Wang F. Development of RGD-based radiotracers for tumor imaging and therapy: translating from bench to bedside. Curr Mol Med. 2013;13:1487–505.

    Article  CAS  PubMed  Google Scholar 

  40. Torchilin VP, editor. Handbook of targeted delivery of imaging agents. Boca Raton: CRC Press; 1995.

Download references

Compliance with ethical standards

Funding

This study was funded by the University of Wisconsin–Madison, the Department of Defense (W81XWH-11-1-0644 & W81XWH-11-1-0648), the National Science Foundation (DGE-1256259), the National Institutes of Health (NIBIB/NCI 1R01CA169365, P30CA014520, T32CA009206, and 5T32GM08349), the US Department of States sponsored Fulbright Scholar Program (1831/FNPDR/2013), and the American Cancer Society (125246-RSG-13-099-01-CCE).

Conflicts of interest

Andrzej Czerwinski and Francisco Valenzuela are employees of Peptides International, Inc. The other authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the University of Wisconsin Institutional Animal Care and Use Committee.

This article does not describe any studies with human participants performed by any of the authors.

Author information

Authors and Affiliations

  1. Department of Medical Physics, University of Wisconsin, Madison, WI, 53705, USA

    Reinier Hernandez, Stephen A. Graves, Robert J. Nickles & Weibo Cai

  2. Peptides International, Inc., Louisville, KY, 40299, USA

    Andrzej Czerwinski & Francisco Valenzuela

  3. Department of Radiology, University of Wisconsin, Madison, WI, 53705, USA

    Rubel Chakravarty, Yunan Yang, Christopher G. England & Weibo Cai

  4. University of Wisconsin Carbone Cancer Center, Madison, WI, 53705, USA

    Weibo Cai

Authors
  1. Reinier Hernandez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrzej Czerwinski
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rubel Chakravarty
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stephen A. Graves
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yunan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christopher G. England
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Robert J. Nickles
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Francisco Valenzuela
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Weibo Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Weibo Cai.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 483 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hernandez, R., Czerwinski, A., Chakravarty, R. et al. Evaluation of two novel 64Cu-labeled RGD peptide radiotracers for enhanced PET imaging of tumor integrin αvβ3 . Eur J Nucl Med Mol Imaging 42, 1859–1868 (2015). https://doi.org/10.1007/s00259-015-3085-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00259-015-3085-7

Keywords

Search

Navigation