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Open Access 01-12-2018 | Research Article

Radiolabeled F(ab′)₂-cetuximab for theranostic purposes in colorectal and skin tumor-bearing mice models

Authors: P.-S. Bellaye, M. Moreau, O. Raguin, A. Oudot, C. Bernhard, J.-M. Vrigneaud, L. Dumont, D. Vandroux, F. Denat, A. Cochet, F. Brunotte, B. Collin

Published in: Clinical and Translational Oncology | Issue 12/2018

Abstract

Purpose

This study aimed to investigate theranostic strategies in colorectal and skin cancer based on fragments of cetuximab, an anti-EGFR mAb, labeled with radionuclide with imaging and therapeutic properties, ¹¹¹In and ¹⁷⁷Lu, respectively.

Methods

We designed F(ab′)₂-fragments of cetuximab radiolabeled with ¹¹¹In and ¹⁷⁷Lu. ¹¹¹In-F(ab′)₂-cetuximab tumor targeting and biodistribution were evaluated by SPECT in BalbC nude mice bearing primary colorectal tumors. The efficacy of ¹¹¹In-F(ab′)₂-cetuximab to assess therapy efficacy was performed on BalbC nude mice bearing colorectal tumors receiving 17-DMAG, an HSP90 inhibitor. Therapeutic efficacy of the radioimmunotherapy based on ¹⁷⁷Lu-F(ab′)₂-cetuximab was evaluated in SWISS nude mice bearing A431 tumors.

Results

Radiolabeling procedure did not change F(ab′)₂-cetuximab and cetuximab immunoreactivity nor affinity for HER1 in vitro. ¹¹¹In-DOTAGA-F(ab′)₂-cetuximab exhibited a peak tumor uptake at 24 h post-injection and showed a high tumor specificity determined by a significant decrease in tumor uptake after the addition of an excess of unlabeled-DOTAGA-F(ab′)₂-cetuximab. SPECT imaging of ¹¹¹In-DOTAGA-F(ab′)₂-cetuximab allowed an accurate evaluation of tumor growth and successfully predicted the decrease in tumor growth induced by 17-DMAG. Finally, ¹⁷⁷Lu-DOTAGA-F(ab′)₂-cetuximab radioimmunotherapy showed a significant reduction of tumor growth at 4 and 8 MBq doses.

Conclusions

¹¹¹In-DOTAGA-F(ab′)₂-cetuximab is a reliable and stable tool for specific in vivo tumor targeting and is suitable for therapy efficacy assessment. ¹⁷⁷Lu-DOTAGA-F(ab′)₂-cetuximab is an interesting theranostic tool allowing therapy and imaging.

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Herbst RS. Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys. 2004;59(2 Suppl):21–6. https://doi.org/10.1016/j.ijrobp.2003.11.041.CrossRefPubMed

Humblet Y. Cetuximab: an IgG(1) monoclonal antibody for the treatment of epidermal growth factor receptor-expressing tumours. Expert Opin Pharmacother. 2004;5(7):1621–33. https://doi.org/10.1517/14656566.5.7.1621.CrossRefPubMed

Bussink J, van der Kogel AJ, Kaanders JH. Activation of the PI3-K/AKT pathway and implications for radioresistance mechanisms in head and neck cancer. Lancet Oncol. 2008;9(3):288–96. https://doi.org/10.1016/S1470-2045(08)70073-1.CrossRefPubMed

Yamamoto VN, Thylur DS, Bauschard M, Schmale I, Sinha UK. Overcoming radioresistance in head and neck squamous cell carcinoma. Oral Oncol. 2016;63:44–51. https://doi.org/10.1016/j.oraloncology.2016.11.002.CrossRefPubMed

Bonner JA, Harari PM, Giralt J, Azarnia N, Shin DM, Cohen RB, et al. Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl j Med. 2006;354(6):567–78. https://doi.org/10.1056/NEJMoa053422.CrossRefPubMed

Petrelli F, Coinu A, Riboldi V, Borgonovo K, Ghilardi M, Cabiddu M, et al. Concomitant platinum-based chemotherapy or cetuximab with radiotherapy for locally advanced head and neck cancer: a systematic review and meta-analysis of published studies. Oral Oncol. 2014;50(11):1041–8. https://doi.org/10.1016/j.oraloncology.2014.08.005.CrossRefPubMed

Ang KK, Zhang Q, Rosenthal DI, Nguyen-Tan PF, Sherman EJ, Weber RS, et al. Randomized phase III trial of concurrent accelerated radiation plus cisplatin with or without cetuximab for stage III to IV head and neck carcinoma: RTOG 0522. J clin oncol off J Am Soc Clin Oncol. 2014;32(27):2940–50. https://doi.org/10.1200/JCO.2013.53.5633.CrossRef

James ML, Gambhir SS. A molecular imaging primer: modalities, imaging agents, and applications. Physiol Rev. 2012;92(2):897–965. https://doi.org/10.1152/physrev.00049.2010.CrossRefPubMed

Milenic DE, Wong KJ, Baidoo KE, Ray GL, Garmestani K, Williams M, et al. Cetuximab: preclinical evaluation of a monoclonal antibody targeting EGFR for radioimmunodiagnostic and radioimmunotherapeutic applications. Cancer Biother Radiopharm. 2008;23(5):619–31. https://doi.org/10.1089/cbr.2008.0493.CrossRefPubMedPubMedCentral

10.

Huhtala T, Laakkonen P, Sallinen H, Yla-Herttuala S, Narvanen A. In vivo SPECT/CT imaging of human orthotopic ovarian carcinoma xenografts with ¹¹¹In-labeled monoclonal antibodies. Nucl Med Biol. 2010;37(8):957–64. https://doi.org/10.1016/j.nucmedbio.2010.03.001.CrossRefPubMed

11.

van Dijk LK, Hoeben BA, Stegeman H, Kaanders JH, Franssen GM, Boerman OC, et al. ¹¹¹In-cetuximab-F(ab’)₂ SPECT imaging for quantification of accessible epidermal growth factor receptors (EGFR) in HNSCC xenografts. Radiother Oncol J Eur Soc Thera Radiol Oncol. 2013;108(3):484–8. https://doi.org/10.1016/j.radonc.2013.06.034.CrossRef

12.

Shih YH, Peng CL, Lee SY, Chiang PF, Yao CJ, Lin WJ, et al. ¹¹¹In-cetuximab as a diagnostic agent by accessible epidermal growth factor (EGF) receptor targeting in human metastatic colorectal carcinoma. Oncotarget. 2015;6(18):16601–10. https://doi.org/10.18632/oncotarget.3968.CrossRefPubMedPubMedCentral

13.

Lee SY, Hong YD, Kim HS, Choi SJ. Synthesis and application of a novel cysteine-based DTPA-NCS for targeted radioimmunotherapy. Nucl Med Biol. 2013;40(3):424–9. https://doi.org/10.1016/j.nucmedbio.2012.12.007.CrossRefPubMed

14.

Brouwers AH, van Eerd JE, Frielink C, Oosterwijk E, Oyen WJ, Corstens FH, et al. Optimization of radioimmunotherapy of renal cell carcinoma: labeling of monoclonal antibody cG250 with ¹³¹I, ⁹⁰Y, ¹⁷⁷Lu, or ¹⁸⁶Re. J Nucl Med Off Publ Soc Nucl Med. 2004;45(2):327–37.

15.

Liu Z, Ma T, Liu H, Jin Z, Sun X, Zhao H, et al. ¹⁷⁷Lu-labeled antibodies for EGFR-targeted SPECT/CT imaging and radioimmunotherapy in a preclinical head and neck carcinoma model. Mol Pharm. 2014;11(3):800–7. https://doi.org/10.1021/mp4005047.CrossRefPubMed

16.

Song IH, Lee TS, Park YS, Lee JS, Lee BC, Moon BS, et al. Immuno-PET Imaging and Radioimmunotherapy of ⁶⁴Cu-/¹⁷⁷Lu-Labeled Anti-EGFR Antibody in Esophageal Squamous Cell Carcinoma Model. J Nucl Med Off Publ Soc Nucl Med. 2016;57(7):1105–11. https://doi.org/10.2967/jnumed.115.167155.CrossRef

17.

Perk LR, Visser GW, Vosjan MJ, Stigter-van Walsum M, Tijink BM, Leemans CR, et al. ⁸⁹Zr as a PET surrogate radioisotope for scouting biodistribution of the therapeutic radiometals ⁹⁰Y and ¹⁷⁷Lu in tumor-bearing nude mice after coupling to the internalizing antibody cetuximab. J Nucl Med Off Publ Soc Nucl Med. 2005;46(11):1898–906.

18.

Harrison A, Walker CA, Parker D, Jankowski KJ, Cox JP, Craig AS, et al. The in vivo release of ⁹⁰Y from cyclic and acyclic ligand-antibody conjugates. Int J Radiat Appl Instrum Part B Nucl Med Biol. 1991;18(5):469–76.CrossRef

19.

Camera L, Kinuya S, Garmestani K, Wu C, Brechbiel MW, Pai LH, et al. Evaluation of the serum stability and in vivo biodistribution of CHX-DTPA and other ligands for yttrium labeling of monoclonal antibodies. J Nucl Med Off Publ Soc Nucl Med. 1994;35(5):882–9.

20.

Smith-Jones PM, Vallabahajosula S, Goldsmith SJ, Navarro V, Hunter CJ, Bastidas D, et al. In vitro characterization of radiolabeled monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen. Can Res. 2000;60(18):5237–43.

21.

Milenic DE, Garmestani K, Chappell LL, Dadachova E, Yordanov A, Ma D, et al. In vivo comparison of macrocyclic and acyclic ligands for radiolabeling of monoclonal antibodies with ¹⁷⁷Lu for radioimmunotherapeutic applications. Nucl Med Biol. 2002;29(4):431–42.CrossRefPubMed

22.

Bernhard C, Moreau M, Lhenry D, Goze C, Boschetti F, Rousselin Y, et al. DOTAGA-anhydride: a valuable building block for the preparation of DOTA-like chelating agents. Chemistry. 2012;18(25):7834–41. https://doi.org/10.1002/chem.201200132.CrossRefPubMed

23.

Moreau M, Raguin O, Vrigneaud JM, Collin B, Bernhard C, Tizon X, et al. DOTAGA-trastuzumab. a new antibody conjugate targeting HER2/Neu antigen for diagnostic purposes. Bioconjug Chem. 2012;23(6):1181–8. https://doi.org/10.1021/bc200680x.CrossRefPubMed

24.

Adams GP, Schier R, McCall AM, Simmons HH, Horak EM, Alpaugh RK, et al. High affinity restricts the localization and tumor penetration of single-chain fv antibody molecules. Can Res. 2001;61(12):4750–5.

25.

Lunt SJ, Chaudary N, Hill RP. The tumor microenvironment and metastatic disease. Clin Exp Metas. 2009;26(1):19–34. https://doi.org/10.1007/s10585-008-9182-2.CrossRef

26.

Wong KJ, Baidoo KE, Nayak TK, Garmestani K, Brechbiel MW, Milenic DE. In Vitro and In Vivo pre-clinical analysis of a F(ab’)² fragment of panitumumab for molecular imaging and therapy of HER1 positive cancers. EJNMMI Res. 2011;1(1):1. https://doi.org/10.1186/2191-219x-1-1.CrossRefPubMedPubMedCentral

27.

Ahsan A, Ramanand SG, Whitehead C, Hiniker SM, Rehemtulla A, Pratt WB, et al. Wild-type EGFR is stabilized by direct interaction with HSP90 in cancer cells and tumors. Neoplasia. 2012;14(8):670–7.CrossRefPubMedPubMedCentral

28.

Zhang F, Wang S, Yin L, Yang Y, Guan Y, Wang W, et al. Quantification of epidermal growth factor receptor expression level and binding kinetics on cell surfaces by surface plasmon resonance imaging. Anal Chem. 2015;87(19):9960–5. https://doi.org/10.1021/acs.analchem.5b02572.CrossRefPubMedPubMedCentral

29.

Marmol I, Sanchez-de-Diego C, Pradilla Dieste A, Cerrada E, Rodriguez Yoldi MJ. Colorectal carcinoma: a general overview and future perspectives in colorectal cancer. Int J Mol Sci. 2017;18(1):197. https://doi.org/10.3390/ijms18010197.CrossRefPubMedCentral

30.

Curtin JC. Novel drug discovery opportunities for colorectal cancer. Expert Opin Drug Discov. 2013;8(9):1153–64. https://doi.org/10.1517/17460441.2013.807249.CrossRefPubMed

31.

Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin. 2013;63(1):11–30. https://doi.org/10.3322/caac.21166.CrossRefPubMed

32.

Khelwatty SA, Essapen S, Seddon AM, Modjtahedi H. Prognostic significance and targeting of HER family in colorectal cancer. Front Biosci. 2013;18:394–421.CrossRef

33.

Harding J, Burtness B. Cetuximab: an epidermal growth factor receptor chemeric human-murine monoclonal antibody. Drugs Today. 2005;41(2):107–27. https://doi.org/10.1358/dot.2005.41.2.882662.CrossRef

34.

Sihver W, Pietzsch J, Krause M, Baumann M, Steinbach J, Pietzsch HJ. Radiolabeled cetuximab conjugates for EGFR targeted cancer diagnostics and therapy. Pharmaceuticals. 2014;7(3):311–38. https://doi.org/10.3390/ph7030311.CrossRefPubMedPubMedCentral

35.

Covell DG, Barbet J, Holton OD, Black CD, Parker RJ, Weinstein JN. Pharmacokinetics of monoclonal immunoglobulin G₁, F(ab’)₂, and Fab’ in mice. Can Res. 1986;46(8):3969–78.

36.

van Dijk LK, Yim CB, Franssen GM, Kaanders JH, Rajander J, Solin O, et al. PET of EGFR with ⁶⁴Cu-cetuximab-F(ab’)² in mice with head and neck squamous cell carcinoma xenografts. Contrast Media Mol Imaging. 2016;11(1):65–70. https://doi.org/10.1002/cmmi.1659.CrossRefPubMed

37.

van Dijk LK, Hoeben BA, Kaanders JH, Franssen GM, Boerman OC, Bussink J. Imaging of epidermal growth factor receptor expression in head and neck cancer with SPECT/CT and ¹¹¹In-labeled cetuximab-F(ab’)₂. J Nucl Med Off Publ Soc Nucl Med. 2013;54(12):2118–24. https://doi.org/10.2967/jnumed.113.123612.CrossRef

38.

Kim DH, Zhou K, Kim DK, Park S, Noh J, Kwon Y, et al. Analysis of interactions between the epidermal growth Factor receptor and soluble ligands on the basis of single-molecule diffusivity in the membrane of living cells. Angew Chem. 2015;54(24):7028–32. https://doi.org/10.1002/anie.201500871.CrossRef

39.

Haeder M, Rotsch M, Bepler G, Hennig C, Havemann K, Heimann B, et al. Epidermal growth factor receptor expression in human lung cancer cell lines. Can Res. 1988;48(5):1132–6.

40.

Yoshida H, Mochizuki M, Kainouchi M, Ishida T, Sakata K, Yokoyama S, et al. Clinical application of indium-111 antimyosin antibody and thallium-201 dual nuclide single photon emission computed tomography in acute myocardial infarction. Ann Nucl Med. 1991;5(1):41–6.CrossRefPubMed

41.

Divgi CR, Welt S, Kris M, Real FX, Yeh SD, Gralla R, et al. Phase I and imaging trial of indium 111-labeled anti-epidermal growth factor receptor monoclonal antibody 225 in patients with squamous cell lung carcinoma. J Natl Cancer Inst. 1991;83(2):97–104.CrossRefPubMed

42.

Brechbiel MW. Bifunctional chelates for metal nuclides. The Q J Nucl Med Mol Imaging Official Publ Ital Assoc Nucl Med. 2008;52(2):166–73.

43.

Weineisen M, Simecek J, Schottelius M, Schwaiger M, Wester HJ. Synthesis and preclinical evaluation of DOTAGA-conjugated PSMA ligands for functional imaging and endoradiotherapy of prostate cancer. EJNMMI Res. 2014;4(1):63. https://doi.org/10.1186/s13550-014-0063-1.CrossRefPubMedPubMedCentral

44.

Baum RP, Kulkarni HR, Schuchardt C, Singh A, Wirtz M, Wiessalla S, et al. ¹⁷⁷Lu-labeled prostate-specific membrane antigen radioligand therapy of metastatic castration-resistant prostate cancer: safety and efficacy. J Nucl Med Off Publ Soc Nucl Med. 2016;57(7):1006–13. https://doi.org/10.2967/jnumed.115.168443.CrossRef

45.

Weineisen M, Schottelius M, Simecek J, Baum RP, Yildiz A, Beykan S, et al. ⁶⁸Ga- and ¹⁷⁷Lu-labeled PSMA I&T: optimization of a PSMA-targeted theranostic concept and first proof-of-concept human studies. J Nucl Med Off Publ Soc Nucl Med. 2015;56(8):1169–76. https://doi.org/10.2967/jnumed.115.158550.CrossRef

46.

Bergsma H, Konijnenberg MW, Kam BL, Teunissen JJ, Kooij PP, de Herder WW, et al. Subacute haematotoxicity after PRRT with ¹⁷⁷Lu-DOTA-octreotate: prognostic factors, incidence and course. Eur J Nucl Med Mol Imaging. 2016;43(3):453–63. https://doi.org/10.1007/s00259-015-3193-4.CrossRefPubMed

47.

Ahmadzadehfar H, Rahbar K, Kurpig S, Bogemann M, Claesener M, Eppard E, et al. Early side effects and first results of radioligand therapy with ¹⁷⁷Lu-DKFZ-617 PSMA of castrate-resistant metastatic prostate cancer: a two-centre study. EJNMMI Res. 2015;5(1):114. https://doi.org/10.1186/s13550-015-0114-2.CrossRefPubMed

48.

van Dijk LK, Boerman OC, Franssen GM, Kaanders JH, Bussink J. ¹¹¹In-cetuximab-F(ab’)₂ SPECT and ¹⁸F-FDG PET for prediction and response monitoring of combined-modality treatment of human head and neck carcinomas in a mouse model. J Nucl Med Off Publ Soc Nucl Med. 2015;56(2):287–92. https://doi.org/10.2967/jnumed.114.148296.CrossRef

49.

Spiegelberg D, Mortensen AC, Selvaraju RK, Eriksson O, Stenerlow B, Nestor M. Molecular imaging of EGFR and CD44v6 for prediction and response monitoring of HSP90 inhibition in an in vivo squamous cell carcinoma model. Eur J Nucl Med Mol Imaging. 2016;43(5):974–82. https://doi.org/10.1007/s00259-015-3260-x.CrossRefPubMed

50.

Borjesson PK, Postema EJ, de Bree R, Roos JC, Leemans CR, Kairemo KJ, et al. Radioimmunodetection and radioimmunotherapy of head and neck cancer. Oral Oncol. 2004;40(8):761–72. https://doi.org/10.1016/j.oraloncology.2003.11.009.CrossRefPubMed

51.

Fischer E, Grunberg J, Cohrs S, Hohn A, Waldner-Knogler K, Jeger S, et al. L1-CAM-targeted antibody therapy and ¹⁷⁷Lu-radioimmunotherapy of disseminated ovarian cancer. Int J Cancer. 2012;130(11):2715–21. https://doi.org/10.1002/ijc.26321.CrossRefPubMed

52.

Tagawa ST, Milowsky MI, Morris M, Vallabhajosula S, Christos P, Akhtar NH, et al. Phase II study of Lutetium-177-labeled anti-prostate-specific membrane antigen monoclonal antibody J591 for metastatic castration-resistant prostate cancer. Clin Cancer Res Off J Am Assoc Cancer Res. 2013;19(18):5182–91. https://doi.org/10.1158/1078-0432.CCR-13-0231.CrossRef

53.

de Cuba EM, Kwakman R, van Egmond M, Bosch LJ, Bonjer HJ, Meijer GA, et al. Understanding molecular mechanisms in peritoneal dissemination of colorectal cancer: future possibilities for personalised treatment by use of biomarkers. Virchows Archiv Int J Pathol. 2012;461(3):231–43. https://doi.org/10.1007/s00428-012-1287-y.CrossRef

Title: Radiolabeled F(ab′)2-cetuximab for theranostic purposes in colorectal and skin tumor-bearing mice models
Authors: P.-S. Bellaye
M. Moreau
O. Raguin
A. Oudot
C. Bernhard
J.-M. Vrigneaud
L. Dumont
D. Vandroux
F. Denat
A. Cochet
F. Brunotte
B. Collin
Publication date: 01-12-2018
Publisher: Springer International Publishing
Published in: Clinical and Translational Oncology / Issue 12/2018
Print ISSN: 1699-048X
Electronic ISSN: 1699-3055
DOI: https://doi.org/10.1007/s12094-018-1886-4

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