Top

European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-07-2015 | Original Article

ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment

Authors: Sixiang Shi, Hao Hong, Hakan Orbay, Stephen A. Graves, Yunan Yang, Jakob D. Ohman, Bai Liu, Robert J. Nickles, Hing C. Wong, Weibo Cai

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 8/2015

Abstract

Purpose

To date, there is no effective therapy for triple-negative breast cancer (TNBC), which has a dismal clinical outcome. Upregulation of tissue factor (TF) expression leads to increased patient morbidity and mortality in many solid tumor types, including TNBC. Our goal was to employ the Fab fragment of ALT-836, a chimeric anti-human TF mAb, for PET imaging of TNBC, which can be used to guide future TNBC therapy.

Methods

ALT-836-Fab was generated by enzymatic papain digestion. SDS-PAGE and FACS studies were performed to evaluate the integrity and TF binding affinity of ALT-836-Fab before NOTA conjugation and ⁶⁴Cu-labeling. Serial PET imaging and biodistribution studies were carried out to evaluate the tumor targeting efficacy and pharmacokinetics in the MDA-MB-231 TNBC model, which expresses high levels of TF on the tumor cells. Blocking studies, histological assessment, as well as RT-PCR were performed to confirm TF specificity of ⁶⁴Cu-NOTA-ALT-836-Fab.

Results

ALT-836-Fab was produced with high purity, which exhibited superb TF binding affinity and specificity. Serial PET imaging revealed rapid and persistent tumor uptake of ⁶⁴Cu-NOTA-ALT-836-Fab (5.1 ± 0.5 %ID/g at 24 h post-injection; n = 4) and high tumor/muscle ratio (7.0 ± 1.2 at 24 h post-injection; n = 4), several-fold higher than that of the blocking group and tumor models that do not express significant level of TF, which was confirmed by biodistribution studies. TF specificity of the tracer was also validated by histology and RT-PCR.

Conclusion

⁶⁴Cu-NOTA-ALT-836-Fab exhibited prominent tissue factor targeting efficiency in MDA-MB-231 TNBC model. The use of a Fab fragment led to fast tumor uptake and good tissue/muscle ratio, which may be translated into same-day immunoPET imaging in the clinical setting to improve TNBC patient management.

Foulkes WD, Smith IE, Reis-Filho JS. Triple-negative breast cancer. N Engl J Med. 2010;363:1938–48.PubMedCrossRef

van Scheltinga AG T, Berghuis P, Nienhuis HH, Timmer-Bosscha H, Pot L, Gaykema SB, et al. Visualising dual downregulation of insulin-like growth factor receptor-1 and vascular endothelial growth factor-A by heat shock protein 90 inhibition effect in triple negative breast cancer. Eur J Cancer. 2014;50:2508–16.CrossRef

Carey L, Winer E, Viale G, Cameron D, Gianni L. Triple-negative breast cancer: disease entity or title of convenience? Nat Rev Clin Oncol. 2010;7:683–92.PubMedCrossRef

Callander NS, Varki N, Rao LV. Immunohistochemical identification of tissue factor in solid tumors. Cancer. 1992;70:1194–201.PubMedCrossRef

Vrana JA, Stang MT, Grande JP, Getz MJ. Expression of tissue factor in tumor stroma correlates with progression to invasive human breast cancer: paracrine regulation by carcinoma cell-derived members of the transforming growth factor beta family. Cancer Res. 1996;56:5063–70.PubMed

Jiang X, Bailly MA, Panetti TS, Cappello M, Konigsberg WH, Bromberg ME. Formation of tissue factor-factor VIIa-factor Xa complex promotes cellular signaling and migration of human breast cancer cells. J Thromb Haemost. 2004;2:93–101.PubMedCrossRef

Contrino J, Hair G, Kreutzer DL, Rickles FR. In situ detection of tissue factor in vascular endothelial cells: correlation with the malignant phenotype of human breast disease. Nat Med. 1996;2:209–15.PubMedCrossRef

Ruf W, Yokota N, Schaffner F. Tissue factor in cancer progression and angiogenesis. Thromb Res. 2010;125 Suppl 2:S36–8.PubMedCrossRef

Ryden L, Grabau D, Schaffner F, Jonsson PE, Ruf W, Belting M. Evidence for tissue factor phosphorylation and its correlation with protease-activated receptor expression and the prognosis of primary breast cancer. Int J Cancer. 2010;126:2330–40.PubMedCentralPubMed

10.

Tesselaar ME, Romijn FP, Van Der Linden IK, Prins FA, Bertina RM, Osanto S. Microparticle-associated tissue factor activity: a link between cancer and thrombosis? J Thromb Haemost. 2007;5:520–7.PubMedCrossRef

11.

Ueno T, Toi M, Koike M, Nakamura S, Tominaga T. Tissue factor expression in breast cancer tissues: its correlation with prognosis and plasma concentration. Br J Cancer. 2000;83:164–70.PubMedCentralPubMedCrossRef

12.

Hong H, Zhang Y, Nayak TR, Engle JW, Wong HC, Liu B, et al. Immuno-PET of tissue factor in pancreatic cancer. J Nucl Med. 2012;53:1748–54.PubMedCentralPubMedCrossRef

13.

Jiao JA, Kelly AB, Marzec UM, Nieves E, Acevedo J, Burkhardt M, et al. Inhibition of acute vascular thrombosis in chimpanzees by an anti-human tissue factor antibody targeting the factor X binding site. Thromb Haemost. 2010;103:224–33.PubMedCentralPubMedCrossRef

14.

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

15.

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

16.

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

17.

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–3003s.PubMed

18.

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

19.

Zhang Y, Hong H, Orbay H, Valdovinos HF, Nayak TR, Theuer CP, et al. PET imaging of CD105/endoglin expression with a 61/64Cu-labeled Fab antibody fragment. Eur J Nucl Med Mol Imaging. 2013;40:759–67.PubMedCentralPubMedCrossRef

20.

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

21.

Chen F, Nayak TR, Goel S, Valdovinos HF, Hong H, Theuer CP, et al. In vivo tumor vasculature targeted PET/NIRF imaging with TRC105(Fab)-conjugated, dual-labeled mesoporous silica nanoparticles. Mol Pharm. 2014;11:4007–14.PubMedCentralPubMedCrossRef

22.

Hewett PW, Murray C. Modulation of human endothelial cell procoagulant activity in tumour models in vitro. Int J Cancer. 1996;66:784–9.PubMedCrossRef

23.

Hjortoe GM, Petersen LC, Albrektsen T, Sorensen BB, Norby PL, Mandal SK, et al. Tissue factor-factor VIIa-specific up-regulation of IL-8 expression in MDA-MB-231 cells is mediated by PAR-2 and results in increased cell migration. Blood. 2004;103:3029–37.PubMedCentralPubMedCrossRef

24.

Zhou J, Stenlinder M, Linder S. Tissue factor mRNA expression in breast carcinoma cells. Oncol Rep. 1996;3:443–5.PubMed

25.

Zhou JN, Ljungdahl S, Shoshan MC, Swedenborg J, Linder S. Activation of tissue-factor gene expression in breast carcinoma cells by stimulation of the RAF-ERK signaling pathway. Mol Carcinog. 1998;21:234–43.PubMedCrossRef

26.

Orbay H, Zhang Y, Hong H, Hacker TA, Valdovinos HF, Zagzebski JA, et al. Positron emission tomography imaging of angiogenesis in a murine hindlimb ischemia model with 64Cu-labeled TRC105. Mol Pharm. 2013;10:2749–56.PubMedCentralPubMedCrossRef

27.

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.PubMedCentralPubMedCrossRef

28.

Ademuyiwa FO, Ellis MJ, Ma CX. Neoadjuvant therapy in operable breast cancer: application to triple negative breast cancer. J Oncol. 2013;2013:219869.PubMedCentralPubMedCrossRef

29.

Hudis CA, Gianni L. Triple-negative breast cancer: an unmet medical need. Oncologist. 2011;16 Suppl 1:1–11.PubMedCrossRef

30.

Schmadeka R, Harmon BE, Singh M. Triple-negative breast carcinoma: current and emerging concepts. Am J Clin Pathol. 2014;141:462–77.PubMedCrossRef

31.

Tchou J, Sonnad SS, Bergey MR, Basu S, Tomaszewski J, Alavi A, et al. Degree of tumor FDG uptake correlates with proliferation index in triple negative breast cancer. Mol Imaging Biol. 2010;12:657–62.PubMedCrossRef

32.

Hama Y, Nakagawa K. Early distant relapse in early stage triple-negative breast cancer: usefulness of FDG-PET for diagnosis of distant metastases. Breast Cancer. 2013;20:191–3.PubMedCrossRef

33.

Dogan BE, Turnbull LW. Imaging of triple-negative breast cancer. Ann Oncol. 2012;23 Suppl 6:vi23–9.PubMedCrossRef

34.

Finn RS, Dering J, Ginther C, Wilson CA, Glaspy P, Tchekmedyian N, et al. Dasatinib, an orally active small molecule inhibitor of both the src and abl kinases, selectively inhibits growth of basal-type/”triple-negative” breast cancer cell lines growing in vitro. Breast Cancer Res Treat. 2007;105:319–26.PubMedCrossRef

35.

Chakravarty R, Goel S, Valdovinos HF, Hernandez R, Hong H, Nickles RJ, et al. Matching the decay half-life with the biological half-life: ImmunoPET imaging with (44)Sc-labeled cetuximab Fab fragment. Bioconjug Chem. 2014;25:2197–204.PubMedCrossRef

36.

Shi S, Yang K, Hong H, Valdovinos HF, Nayak TR, Zhang Y, et al. Tumor vasculature targeting and imaging in living mice with reduced graphene oxide. Biomaterials. 2013;34:3002–9.PubMedCentralPubMedCrossRef

Title: ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment
Authors: Sixiang Shi
Hao Hong
Hakan Orbay
Stephen A. Graves
Yunan Yang
Jakob D. Ohman
Bai Liu
Robert J. Nickles
Hing C. Wong
Weibo Cai
Publication date: 01-07-2015
Publisher: Springer Berlin Heidelberg
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 8/2015
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-015-3038-1

Keynote webinar | Spotlight on medication adherence

Springer Medicine

ImmunoPET of tissue factor expression in triple-negative breast cancer with a radiolabeled antibody Fab fragment

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 8/2015

Tumour targeting and radiation dose of radioimmunotherapy with 90Y-rituximab in CD20+ B-cell lymphoma as predicted by 89Zr-rituximab immuno-PET: impact of preloading with unlabelled rituximab

Preoperative risk stratification using metabolic parameters of 18F-FDG PET/CT in patients with endometrial cancer

Thoracic staging of non-small-cell lung cancer using integrated 18F-FDG PET/MR imaging: diagnostic value of different MR sequences

V/P SPECT as a diagnostic tool for pregnant women with suspected pulmonary embolism

Radioembolization with 90Y-loaded microspheres: high clinical impact of treatment simulation with MAA-based dosimetry

Towards tailored radiopeptide therapy