nach oben

European Journal of Nuclear Medicine and Molecular Imaging

Erschienen in:

01.05.2013 | Original Article

PET imaging of CD105/endoglin expression with a ^61/64Cu-labeled Fab antibody fragment

verfasst von: Yin Zhang, Hao Hong, Hakan Orbay, Hector F. Valdovinos, Tapas R. Nayak, Charles P. Theuer, Todd E. Barnhart, Weibo Cai

Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging | Ausgabe 5/2013

Einloggen, um Zugang zu erhalten

Abstract

Purpose

The goal of this study was to generate and characterize the Fab fragment of TRC105, a monoclonal antibody that binds with high affinity to human and murine CD105 (i.e., endoglin), and investigate its potential for PET imaging of tumor angiogenesis in a small-animal model after ^61/64Cu labeling.

Methods

TRC105-Fab was generated by enzymatic papain digestion. The integrity and CD105 binding affinity of TRC105-Fab was evaluated before NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) conjugation and ^61/64Cu labeling. Serial PET imaging and biodistribution studies were carried out in the syngeneic 4T1 murine breast cancer model to quantify tumor targeting efficiency and normal organ distribution of ^61/64Cu-NOTA-TRC105-Fab. Blocking studies with unlabeled TRC105 were performed to confirm CD105 specificity of the tracer in vivo. Immunofluorescence staining was also conducted to correlate tracer uptake in the tumor and normal tissues with CD105 expression.

Results

TRC105-Fab was produced with high purity through papain digestion of TRC105, as confirmed by SDS-PAGE, HPLC analysis, and mass spectrometry. ^61/64Cu labeling of NOTA-TRC105-Fab was achieved with about 50 % yield (specific activity about 44 GBq/μmol). PET imaging revealed rapid uptake of ⁶⁴Cu-NOTA-TRC105-Fab in the 4T1 tumor (3.6 ± 0.4, 4.2 ± 0.5, 4.9 ± 0.3, 4.4 ± 0.7, and 4.6 ± 0.8 %ID/g at 0.5, 2, 5, 16, and 24 h after injection, respectively; n = 4). Since tumor uptake peaked soon after tracer injection, ⁶¹Cu-labeled TRC105-Fab was also able to provide tumor contrast at 3 and 8 h after injection. CD105 specificity of the tracer was confirmed with blocking studies and histological examination.

Conclusion

We report PET imaging of CD105 expression using ^61/64Cu-NOTA-TRC105-Fab, which exhibited prominent and target-specific uptake in the 4T1 tumor. The use of a Fab fragment led to much faster tumor uptake (which peaked at a few hours after tracer injection) compared to radiolabeled intact antibody, which may be translated into same-day immunoPET imaging for clinical investigation.

Carmeliet P, Jain RK. Angiogenesis in cancer and other diseases. Nature. 2000;407:249–57.PubMedCrossRef

Backer MV, Backer JM. Imaging key biomarkers of tumor angiogenesis. Theranostics. 2012;2:502–15.PubMedCrossRef

Dijkgraaf I, Boerman OC. Radionuclide imaging of tumor angiogenesis. Cancer Biother Radiopharm. 2009;24:637–47.PubMedCrossRef

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

Cai W, Rao J, Gambhir SS, Chen X. How molecular imaging is speeding up anti-angiogenic drug development. Mol Cancer Ther. 2006;5:2624–33.PubMedCrossRef

Backer MV, Levashova Z, Patel V, Jehning BT, Claffey K, Blankenberg FG, et al. Molecular imaging of VEGF receptors in angiogenic vasculature with single-chain VEGF-based probes. Nat Med. 2007;13:504–9.PubMedCrossRef

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

Zhang Y, Yang Y, Hong H, Cai W. Multimodality molecular imaging of CD105 (Endoglin) expression. Int J Clin Exp Med. 2011;4:32–42.PubMed

Cai W, Chen X. Multimodality imaging of vascular endothelial growth factor and vascular endothelial growth factor receptor expression. Front Biosci. 2007;12:4267–79.PubMedCrossRef

10.

Wang RE, Niu Y, Wu H, Amin MN, Cai J. Development of NGR peptide-based agents for tumor imaging. Am J Nucl Med Mol Imaging. 2011;1:36–46.PubMed

11.

Dallas NA, Samuel S, Xia L, Fan F, Gray MJ, Lim SJ, et al. Endoglin (CD105): a marker of tumor vasculature and potential target for therapy. Clin Cancer Res. 2008;14:1931–7.PubMedCrossRef

12.

Fonsatti E, Nicolay HJ, Altomonte M, Covre A, Maio M. Targeting cancer vasculature via endoglin/CD105: a novel antibody-based diagnostic and therapeutic strategy in solid tumours. Cardiovasc Res. 2010;86:12–9.PubMedCrossRef

13.

Hong H, Yang Y, Zhang Y, Engle JW, Barnhart TE, Nickles RJ, et al. Positron emission tomography imaging of CD105 expression during tumor angiogenesis. Eur J Nucl Med Mol Imaging. 2011;38:1335–43.PubMedCrossRef

14.

Rosen LS, Hurwitz HI, Wong MK, Goldman J, Mendelson DS, Figg WD, et al. A phase I first-in-human study of TRC105 (anti-endoglin antibody) in patients with advanced cancer. Clin Cancer Res. 2012;18:4820–9.PubMedCrossRef

15.

Wu AM, Olafsen T. Antibodies for molecular imaging of cancer. Cancer J. 2008;14:191–7.PubMedCrossRef

16.

Andrew SM, Pimm MV, Perkins AC, Baldwin RW. Comparative imaging and biodistribution studies with an anti-CEA monoclonal antibody and its F(ab)2 and Fab fragments in mice with colon carcinoma xenografts. Eur J Nucl Med. 1986;12:168–75.PubMedCrossRef

17.

Hoeben BA, Kaanders JH, Franssen GM, Troost EG, Rijken PF, Oosterwijk E, et al. PET of hypoxia with ⁸⁹Zr-labeled cG250-F(ab’)2 in head and neck tumors. J Nucl Med. 2010;51:1076–83.PubMedCrossRef

18.

Leyton JV, Olafsen T, Lepin EJ, Hahm S, Bauer KB, Reiter RE, et al. Humanized radioiodinated minibody for imaging of prostate stem cell antigen-expressing tumors. Clin Cancer Res. 2008;14:7488–96.PubMedCrossRef

19.

Cai W, Olafsen T, Zhang X, Cao Q, Gambhir SS, Williams LE, et al. PET imaging of colorectal cancer in xenograft-bearing mice by use of an ¹⁸F-labeled T84.66 anti-carcinoembryonic antigen diabody. J Nucl Med. 2007;48:304–10.PubMedCrossRef

20.

Olafsen T, Sirk SJ, Olma S, Shen CK, Wu AM. ImmunoPET using engineered antibody fragments: fluorine-18 labeled diabodies for same-day imaging. Tumour Biol. 2012;33:669–77.PubMedCrossRef

21.

Griffiths GL, Goldenberg DM, Roesch F, Hansen HJ. Radiolabeling of an anti-carcinoembryonic antigen antibody Fab’ fragment (CEA-Scan) with the positron-emitting radionuclide Tc-94m. Clin Cancer Res. 1999;5:3001s–3s.PubMed

22.

Yoshida C, Tsuji AB, Sudo H, Sugyo A, Sogawa C, Inubushi M, et al. Development of positron emission tomography probe of ⁶⁴Cu-labeled anti-C-kit 12A8 Fab to measure protooncogene C-kit expression. Nucl Med Biol. 2011;38:331–7.PubMedCrossRef

23.

Burvenich IJ, Schoonooghe S, Blanckaert P, Bacher K, Vervoort L, Coene E, et al. Biodistribution and planar gamma camera imaging of ¹²³I- and ¹³¹I-labeled F(ab’)2 and Fab fragments of monoclonal antibody 14C5 in nude mice bearing an A549 lung tumor. Nucl Med Biol. 2007;34:257–65.PubMedCrossRef

24.

Brouwers A, Mulders P, Oosterwijk E, Buijs W, Corstens F, Boerman O, et al. Pharmacokinetics and tumor targeting of ¹³¹I-labeled F(ab’)2 fragments of the chimeric monoclonal antibody G250: preclinical and clinical pilot studies. Cancer Biother Radiopharm. 2004;19:466–77.PubMed

25.

Sandstrom K, Haylock AK, Spiegelberg D, Qvarnstrom F, Wester K, Nestor M. A novel CD44v6 targeting antibody fragment with improved tumor-to-blood ratio. Int J Oncol. 2012;40:1525–32.PubMed

26.

Rousseaux J, Rousseaux-Prevost R, Bazin H. Optimal conditions for the preparation of Fab and F(ab’)2 fragments from monoclonal IgG of different rat IgG subclasses. J Immunol Methods. 1983;64:141–6.PubMedCrossRef

27.

Hong H, Severin GW, Yang Y, Engle JW, Zhang Y, Barnhart TE, et al. Positron emission tomography imaging of CD105 expression with ⁸⁹Zr-Df-TRC105. Eur J Nucl Med Mol Imaging. 2012;39:138–48.PubMedCrossRef

28.

Fonsatti E, Jekunen AP, Kairemo KJ, Coral S, Snellman M, Nicotra MR, et al. Endoglin is a suitable target for efficient imaging of solid tumors: in vivo evidence in a canine mammary carcinoma model. Clin Cancer Res. 2000;6:2037–43.PubMed

29.

Zhang Y, Hong H, Engle JW, Yang Y, Theuer CP, Barnhart TE, et al. Positron emission tomography and optical imaging of tumor CD105 expression with a dual-labeled monoclonal antibody. Mol Pharm. 2012;9:645–53.PubMedCrossRef

30.

Zhang Y, Hong H, Engle JW, Yang Y, Barnhart TE, Cai W. Positron emission tomography and near-infrared fluorescence imaging of vascular endothelial growth factor with dual-labeled bevacizumab. Am J Nucl Med Mol Imaging. 2012;2:1–13.PubMed

31.

Zhang Y, Hong H, Severin GW, Engle JW, Yang Y, Goel S, et al. ImmunoPET and near-infrared fluorescence imaging of CD105 expression using a monoclonal antibody dual-labeled with ⁸⁹Zr and IRDye 800CW. Am J Transl Res. 2012;4:333–46.PubMed

32.

Dearling JLJ, Voss SD, Dunning P, Snay E, Fahey F, Smith SV, et al. Imaging cancer using PET – the effect of the bifunctional chelator on the biodistribution of a ⁶⁴Cu-labeled antibody. Nucl Med Biol. 2011;38:29–38.PubMedCrossRef

33.

Hong H, Benink HA, Zhang Y, Yang Y, Uyeda HT, Engle JW, et al. HaloTag: a novel reporter gene for positron emission tomography. Am J Transl Res. 2011;3:392–403.PubMed

34.

Zhang Y, Hong H, Engle JW, Bean J, Yang Y, Leigh BR, et al. Positron emission tomography imaging of CD105 expression with a ⁶⁴Cu-labeled monoclonal antibody: NOTA is superior to DOTA. PLoS One. 2011;6:e28005.PubMedCrossRef

35.

Wu AM, Senter PD. Arming antibodies: prospects and challenges for immunoconjugates. Nat Biotechnol. 2005;23:1137–46.PubMedCrossRef

36.

Seon BK, Haba A, Matsuno F, Takahashi N, Tsujie M, She X, et al. Endoglin-targeted cancer therapy. Curr Drug Deliv. 2011;8:135–43.PubMedCrossRef

37.

Hong H, Zhang Y, Severin GW, Yang Y, Engle JW, Niu G, et al. Multimodality imaging of breast cancer experimental lung metastasis with bioluminescence and a monoclonal antibody dual-labeled with ⁸⁹Zr and IRDye 800CW. Mol Pharm. 2012;9:2339–49.CrossRef

38.

Grassi I, Nanni C, Allegri V, Morigi JJ, Montini GC, Castellucci P, et al. The clinical use of PET with ¹¹C-acetate. Am J Nucl Med Mol Imaging. 2012;2:33–47.PubMed

39.

Vach W, Høilund-Carlsen PF, Fischer BM, Gerke O, Weber W. How to study optimal timing of PET/CT for monitoring of cancer treatment. Am J Nucl Med Mol Imaging. 2011;1:54–62.PubMed

40.

Alauddin MM. Positron emission tomography (PET) imaging with ¹⁸F-based radiotracers. Am J Nucl Med Mol Imaging. 2012;2:55–76.PubMed

41.

Ravetch JV, Bolland S. IgG Fc receptors. Annu Rev Immunol. 2001;19:275–90.PubMedCrossRef

42.

Costello B, Li C, Duff S, Butterworth D, Khan A, Perkins M, et al. Perfusion of ⁹⁹mTc-labeled CD105 Mab into kidneys from patients with renal carcinoma suggests that CD105 is a promising vascular target. Int J Cancer. 2004;109:436–41.PubMedCrossRef

43.

Williams HA, Robinson S, Julyan P, Zweit J, Hastings D. A comparison of PET imaging characteristics of various copper radioisotopes. Eur J Nucl Med Mol Imaging. 2005;32:1473–80.PubMedCrossRef

44.

Tsujie M, Uneda S, Tsai H, Seon BK. Effective anti-angiogenic therapy of established tumors in mice by naked anti-human endoglin (CD105) antibody: differences in growth rate and therapeutic response between tumors growing at different sites. Int J Oncol. 2006;29:1087–94.PubMed

45.

Matsuno F, Haruta Y, Kondo M, Tsai H, Barcos M, Seon BK. Induction of lasting complete regression of preformed distinct solid tumors by targeting the tumor vasculature using two new anti-endoglin monoclonal antibodies. Clin Cancer Res. 1999;5:371–82.PubMed

Titel: PET imaging of CD105/endoglin expression with a 61/64Cu-labeled Fab antibody fragment
verfasst von: Yin Zhang
Hao Hong
Hakan Orbay
Hector F. Valdovinos
Tapas R. Nayak
Charles P. Theuer
Todd E. Barnhart
Weibo Cai
Publikationsdatum: 01.05.2013
Verlag: Springer-Verlag
Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging / Ausgabe 5/2013
Print ISSN: 1619-7070
Elektronische ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-012-2334-2

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusion

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 5/2013

The joint IAEA, EANM, and SNMMI practical guidance on peptide receptor radionuclide therapy (PRRNT) in neuroendocrine tumours

Multiple opportunities to reduce radiation dose from myocardial perfusion imaging

EFOMP and EANM: joint recommendations for a curriculum for the education and training of physicists in nuclear medicine

124I PET/CT in the pretherapeutic staging of differentiated thyroid carcinoma: comparison with posttherapy 131I SPECT/CT

Correlation between [18F]FDG PET/CT and volume perfusion CT in primary tumours and mediastinal lymph nodes of non-small-cell lung cancer

Prognostic implications of volume-based measurements on FDG PET/CT in stage III non-small-cell lung cancer after induction chemotherapy