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European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-12-2007 | Molecular imaging

In vivo imaging of tumour angiogenesis in mice with the α_vβ₃ integrin-targeted tracer ^99mTc-RAFT-RGD

Authors: Lucie Sancey, Valérie Ardisson, Laurent M. Riou, Mitra Ahmadi, Danièle Marti-Batlle, Didier Boturyn, Pascal Dumy, Daniel Fagret, Catherine Ghezzi, Jean-Philippe Vuillez

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 12/2007

Abstract

Purpose

The molecular imaging of tumour neoangiogenesis currently represents a major field of research for the diagnostic and treatment strategy of solid tumours. Endothelial cells from tumour neovessels overexpress the α_vβ₃ integrin, which selectively binds to Arg-Gly-Asp (RGD)-containing peptides. We evaluated the potential of the novel radiotracer ^99mTc-RAFT-RGD for the non-invasive molecular imaging of α_vβ₃ integrin expression in mice models of tumour development.

Methods

^99mTc-RAFT-RGD, ^99mTc-cRGD (specific control) and ^99mTc-RAFT-RAD (non-specific control) were injected intravenously to mice bearing B16F0 or TS/A-pc tumours. In vivo whole-body tomographic imaging and post-mortem biodistribution studies were performed 60 min following tracer injection. Adjacent tumour slices were used to compare the localisation of neovessels from immunostaining and the pattern of ^99mTc-RAFT-RGD uptake from autoradiographic ex vivo imaging.

Results

Biodistribution studies indicated that ^99mTc-RAFT-RGD tumour uptake was significantly higher than that of ^99mTc-RAFT-RAD in B16F0 (2.4±0.5 vs 1.0±0.1%ID/g, respectively) and in TS/A-pc tumours (2.7±0.8 vs 0.7±0.1%ID/g, respectively). Immunohistochemical and autoradiographic studies indicated that ^99mTc-RAFT-RGD intratumoural uptake preferentially occurred in angiogenic areas. Tomographic imaging allowed tumour visualisation following injection of ^99mTc-RAFT-RGD and ^99mTc-cRGD with similar tumour-to-contralateral muscle (T/CM) ratios in B16F0 and in TS/A-pc tumours whereas ^99mTc-RAFT-RAD T/CM ratios did not allow tumour imaging. In accordance with the higher level of α_vβ₃ integrin expression on TS/A-pc tumours than on B16F0 tumours as determined from western blot and immunoprecipitation analyses, the ^99mTc-RAFT-RGD T/CM ratio was significantly higher in TS/A-pc than in B16F0 tumours.

Conclusion

^99mTc-RAFT-RGD allowed the in vivo imaging of α_vβ₃ integrin tumour expression.

Folkman J. Role of angiogenesis in tumor growth and metastasis. Semin Oncol 2002;29(Suppl 16):15–8.PubMed

Beer AJ, Haubner R, Goebel M, Luderschmidt S, Spilker ME, Wester HJ, et al. Biodistribution and pharmacokinetics of the alphavbeta3-selective tracer¹⁸F-galacto-RGD in cancer patients. J Nucl Med 2005;46:1333–41.PubMed

Haubner R, Weber WA, Beer AJ, Vabuliene E, Reim D, Sarbia M, et al. Non-invasive visualization of the activated alphavbeta3 integrin in cancer patients by positron emission tomography and [¹⁸F]-galacto-RGD. PLoS Med 2005;2:244–52.CrossRef

Tucker GC. Integrins: molecular targets in cancer therapy. Curr Oncol Rep 2006;8:96–103.PubMedCrossRef

Folkman J. Tumor angiogenesis: therapeutic implications. N Engl J Med 1971;285:1182–6.PubMedCrossRef

Stupack DG, Cheresh DA. Get a ligand, get a life: integrins, signaling and cell survival. J Cell Sci 2002;115(Pt 19):3729–38.PubMedCrossRef

Rupp PA, Czirok A, Little CD. Alphavbeta3 integrin-dependent endothelial cell dynamics in vivo. Development 2004;131:2887–97.PubMedCrossRef

Ruoslahti E, Pierschbacher MD. New perspectives in cell adhesion: RGD and integrins. Science 1987;238:491–7.PubMedCrossRef

Brooks PC, Clark RA, Cheresh DA. Requirement of vascular integrin alpha v beta 3 for angiogenesis. Science 1994;264:569–71.PubMedCrossRef

10.

Horton MA. The alpha v beta 3 integrin “vitronectin receptor”. Int J Biochem Cell Biol 1997;29:721–5.PubMedCrossRef

11.

Chen X, Park R, Shahinian AH, Bading JR, Conti PS. Pharmacokinetics and tumor retention of [¹²⁵]I-labeled RGD peptide are improved by PEGylation. Nucl Med Biol 2004;31:11–9.PubMedCrossRef

12.

Gruber G, Hess J, Stiefel C, Aebersold DM, Zimmer Y, Greiner RH, et al. Correlation between the tumoral expression of beta3-integrin and outcome in cervical cancer patients who had undergone radiotherapy. Br J Cancer 2005;92:41–6.PubMedCrossRef

13.

Hynes RO. Integrins: versatility, modulation, and signaling in cell adhesion. Cell 1992;69:11–25.PubMedCrossRef

14.

Ruoslahti E. The RGD story: a personal account. Matrix Biology 2003;22:459–65.PubMedCrossRef

15.

Eskens FA, Dumez H, Hoekstra R, Perschl A, Brindley C, Bottcher S, et al. Phase I and pharmacokinetic study of continuous twice weekly intravenous administration of Cilengitide (EMD 121974), a novel inhibitor of the integrins alphavbeta3 and alphavbeta5 in patients with advanced solid tumors. Eur J Cancer 2003;39:917–26.PubMedCrossRef

16.

Raguse JD, Gath HJ, Bier J, Riess H, Oettle H. Cilengitide (EMD 121974) arrests the growth of a heavily pretreated highly vascularised head and neck tumor. Oral Oncol 2004;40:228–30.PubMedCrossRef

17.

Burke PA, DeNardo SJ, Miers LA, Lamborn KR, Matzku S, DeNardo GL. Cilengitide targeting of alpha(v)beta(3) integrin receptor synergizes with radioimmunotherapy to increase efficacy and apoptosis in breast cancer xenografts. Cancer Res 2002;62:4263–72.PubMed

18.

Brooks PC, Montgomery AM, Rosenfeld M, Reisfeld RA, Hu T, Klier G, et al. Integrin alpha v beta 3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell 1994;79:1157–64.PubMedCrossRef

19.

Harris TD, Kalogeropoulos S, Nguyen T, Liu S, Bartis J, Ellars C, et al. Design, synthesis, and evaluation of radiolabeled integrin alpha v beta 3 receptor antagonists for tumor imaging and radiotherapy. Cancer Biother Radiopharm 2003;18:627–41.PubMedCrossRef

20.

Chen X, Park R, Khankaldyyan V, Gonzales-Gomez I, Tohme M, Moats RA, et al. Longitudinal microPET imaging of brain tumor growth with F-18-labeled RGD peptide. Mol Imaging Biol 2006;8:9–15.PubMedCrossRef

21.

Fani M, Psimadas D, Zikos C, Xanthopoulos S, Loudos GK, Bouziotis P, et al. Comparative evaluation of linear and cyclic ^99mTc-RGD peptides for targeting of integrins in tumor angiogenesis. Anticancer Res 2006;26:431–4.PubMed

22.

Castel S, Pagan R, Mitjans F, Piulats J, Goodman S, Jonczyk A, et al. RGD peptides and monoclonal antibodies, antagonists of alpha(v)-integrin, enter the cells by independent endocytic pathways. Lab Invest 2001;81:1615–26.PubMed

23.

Janssen ML, Oyen WJ, Dijkgraaf I, Massuger LF, Frielink C, Edwards DS, et al. Tumor targeting with radiolabeled alpha(v)beta(3) integrin binding peptides in a nude mouse model. Cancer Res 2002;62:6146–51.PubMed

24.

Kok RJ, Schraa AJ, Bos EJ, Moorlag HE, Asgeirsdottir SA, Everts M, et al. Preparation and functional evaluation of RGD-modified proteins as alpha(v)beta(3) integrin directed therapeutics. Bioconjug Chem 2002;13:128–35.PubMedCrossRef

25.

Thumshirn G, Hersel U, Goodman SL, Kessler H. Multimeric cyclic RGD peptides as potential tools for tumor targeting: solid-phase peptide synthesis and chemoselective oxime ligation. Chemistry 2003;9:2717–25.PubMedCrossRef

26.

Garanger E, Boturyn D, Jin Z, Dumy P, Favrot MC, Coll JL. New multifunctional molecular conjugate vector for targeting, imaging, and therapy of tumors. Mol Ther 2005;12:1168–75.PubMedCrossRef

27.

Boturyn D, Dumy P. A convenient access to alpha(V) beta(3)/alpha(V) beta(5) integrin ligand conjugates: regioselective solid-phase functionalisation of an RGD based peptide. Tetrahedron Lett 2001;42:2787–90.CrossRef

28.

Boturyn D, Coll JL, Garanger E, Favrot MC, Dumy P. Template assembled cyclopeptides as multimeric system for integrin targeting and endocytosis. J Am Chem Soc 2004;126:5730–9.PubMedCrossRef

29.

Garanger E, Boturyn D, Coll JL, Favrot MC, Dumy P. Multivalent RGD synthetic peptides as potent alphaVbeta3 integrin ligands. Org Biomol Chem 2006;4:1958–65.PubMedCrossRef

30.

Cowden Dahl KD, Robertson SE, Weaver VM, Simon MC. Hypoxia-inducible factor regulates alphavbeta3 integrin cell surface expression. Mol Biol Cell 2005;16:1901–12.PubMedCrossRef

31.

Denoyer D, Perek N, Le Jeune N, Dubois F. Spectrum of radiopharmaceuticals in nuclear oncology. Curr Cancer Drug Targets 2006;6:181–96.PubMedCrossRef

32.

Haubner R, Wester HJ. Radiolabeled tracers for imaging of tumor angiogenesis and evaluation of anti-angiogenic therapies. Curr Pharm Des 2004;10:1439–55.PubMedCrossRef

33.

Friedlander M, Brooks PC, Shaffer RW, Kincaid CM, Varner JA, Cheresh DA. Definition of two angiogenic pathways by distinct alpha v integrins. Science 1995;270:1500–2.PubMedCrossRef

34.

Pecheur I, Peyruchaud O, Serre CM, Guglielmi J, Voland C, Bourre F, et al. Integrin alpha[v]beta3 expression confers on tumor cells a greater propensity to metastasize to bone. Faseb J 2002;16:1266–8.PubMed

35.

Rolli M, Fransvea E, Pilch J, Saven A, Felding-Habermann B. Activated integrin alphavbeta3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells. Proc Natl Acad Sci U S A 2003;100:9482–7.PubMedCrossRef

36.

Haubner R, Wester HJ, Weber WA, Mang C, Ziegler SI, Goodman SL, et al. Noninvasive imaging of alpha(v)beta(3) integrin expression using F-18-labeled RGD-containing glycopeptide and positron emission tomography. Cancer Res 2001;61:1781–5.PubMed

37.

Jin ZH, Josserand V, Razkin J, Garanger E, Boturyn D, Favrot MC, et al. Noninvasive optical imaging of ovarian metastases using Cy5-labeled RAFT-c(-RGDfK-)4. Mol Imaging 2006;5:188–97.PubMed

38.

Jin ZH, Razkin J, Josserand V, Boturyn D, Grichine A, Texier I, et al. In vivo non-invasive optical imaging of receptor-mediated RGD internalization using self-quenched Cy5-labeled RAFT-c(-RGDfK-)₄. Mol Imaging 2007;6:43–55.PubMed

39.

Dijkgraaf I, Kruijtzer JA, Liu S, Soede AC, Oyen WJ, Corstens FH, et al. Improved targeting of the alpha(v)beta (3) integrin by multimerisation of RGD peptides. Eur J Nucl Med Mol Imaging 2007;34:267–73.PubMedCrossRef

40.

Liu S, Hsieh WY, Jiang Y, Kim YS, Sreerama SG, Chen X, et al. Evaluation of a^99mTc-labeled cyclic RGD tetramer for noninvasive imaging integrin alpha(v)beta3-positive breast cancer. Bioconjug Chem 2007;18:438–46.PubMedCrossRef

41.

Chen X, Tohme M, Park R, Hou Y, Bading JR, Conti PS. Micro-PET imaging of alphavbeta3-integrin expression with¹⁸F-labeled dimeric RGD peptide. Mol Imaging 2004;3:96–104.PubMedCrossRef

42.

Wu Y, Zhang X, Xiong Z, Cheng Z, Fisher DR, Liu S, et al. MicroPET imaging of glioma integrin alphav beta3 expression using [64]Cu-labeled tetrameric RGD peptide. J Nucl Med 2005;46:1707–18.PubMed

43.

Trejtnar F, Laznicek M, Laznickova A, Mather SJ. Pharmacokinetics and renal handling of^99mTc-labeled peptides. J Nucl Med 2000;41:177–82.PubMed

44.

Mulder WJ, Strijkers GJ, Habets JW, Bleeker EJ, van der Schaft DW, Storm G, et al. MR molecular imaging and fluorescence microscopy for identification of activated tumor endothelium using a bimodal lipidic nanoparticle. FASEB J 2005;19:2008–10.PubMed

45.

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

Title: In vivo imaging of tumour angiogenesis in mice with the αvβ3 integrin-targeted tracer 99mTc-RAFT-RGD
Authors: Lucie Sancey
Valérie Ardisson
Laurent M. Riou
Mitra Ahmadi
Danièle Marti-Batlle
Didier Boturyn
Pascal Dumy
Daniel Fagret
Catherine Ghezzi
Jean-Philippe Vuillez
Publication date: 01-12-2007
Publisher: Springer-Verlag
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 12/2007
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-007-0497-z

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

In vivo imaging of tumour angiogenesis in mice with the α_vβ₃ integrin-targeted tracer ^99mTc-RAFT-RGD

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

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