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

  • Sixiang Shi
  • Hao Hong
  • Hakan Orbay
  • Stephen A. Graves
  • Yunan Yang
  • Jakob D. Ohman
  • Bai Liu
  • Robert J. Nickles
  • Hing C. Wong
  • Weibo CaiEmail author
Original Article

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.

Keywords

Triple-negative breast cancer (TNBC) Positron emission tomography (PET) Tissue factor (TF) Antibody fragment Fab 

Notes

Acknowledgments

This work was supported in part by the University of Wisconsin-Madison, the National Institutes of Health (NIBIB/NCI 1R01CA169365, P30CA014520, and T32CA009206), the Department of Defense (W81XWH-11-1-0644 and W81XWH-11-1-0648), and the American Cancer Society (125246-RSG-13-099-01-CCE).

Compliance with ethical standards

Conflicts of interest

Bai Liu and Hing C. Wong are employees of Altor Bioscience Corporation. The other authors declare that they had no conflicts of interest.

Statement of human rights

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

Statement on the welfare of animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Sixiang Shi
    • 1
  • Hao Hong
    • 2
  • Hakan Orbay
    • 2
  • Stephen A. Graves
    • 3
  • Yunan Yang
    • 2
  • Jakob D. Ohman
    • 2
  • Bai Liu
    • 4
  • Robert J. Nickles
    • 3
  • Hing C. Wong
    • 4
  • Weibo Cai
    • 1
    • 2
    • 3
    • 5
    • 6
    Email author
  1. 1.Materials Science ProgramUniversity of WisconsinMadisonUSA
  2. 2.Department of RadiologyUniversity of WisconsinMadisonUSA
  3. 3.Department of Medical PhysicsUniversity of WisconsinMadisonUSA
  4. 4.Altor BioScienceMiramarUSA
  5. 5.University of Wisconsin Carbone Cancer CenterMadisonUSA
  6. 6.Departments of Radiology and Medical PhysicsUniversity of WisconsinMadisonUSA

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