Measuring Response to Therapy By Near-infrared Imaging of Tumors Using A Phosphatidylserine-targeting Antibody Fragment

Authors: Jian Gong, Richard Archer, Michael Brown, Seth Fisher, Connie Chang, Matthew Peacock, Christopher Hughes, Bruce Freimark


Imaging tumors and their response to treatment could be a valuable biomarker toward early assessment of therapy in patients with cancer. Phosphatidylserine (PS) is confined to the inner leaflet of the plasma membrane in normal cells but is externalized on tumor vascular endothelial cells (ECs) and tumor cells, and PS exposure is further enhanced in response to radiation and chemotherapy. In the present study, we evaluated the potential of a PS-targeting human F(ab′)2 antibody fragment, PGN650, to detect exposure of PS in tumor-bearing mice. Tumor uptake of PGN650 was measured by near-infrared optical imaging in human tumor xenografts in immunodeficient mice. PGN650 specifically targeted tumors and was shown to target CD31-positive ECs and tumor cells. Tumor uptake of PGN650 was significantly higher in animals pretreated with docetaxel. The peak tumor to normal tissue (T/N) ratio of probe was observed at 24 hours postinjection of probe, and tumor binding was detected for at least 120 hours. In repeat dose studies, PGN650 uptake in tumors was significantly higher following pretreatment with docetaxel compared to baseline uptake prior to treatment. PGN650 may be a useful probe to detect PS exposed in tumors and to monitor enhanced PS exposure to optimize therapeutic agents to treat tumors.

To access this content please select one of the following options:


Login to an existing account

Claim access now

For current subscribers with Society Membership or Account Number.

  1. Hirsch FR, Franklin WA, Gazdar AF, Bunn PA. Early detection of lung cancer: clinical perspectives of recent advances in biology and radiology. Clin Cancer Res 2001;7:5-22.
  2. Holmgren L, O'Reilly MS, Folkman J. Dormancy of micrometastases: balanced proliferation and apoptosis in the presence of angiogenesis suppression. Nat Med 1995;1:149-53, doi:10.1038/nm0295-149.
  3. Belhocine T, Steinmetz N, Green A, Rigo P. In vivo imaging of chemotherapy-induced apoptosis in human cancers. Ann N Y Acad Sci 2003 1010;:525-9.
  4. Williamson P, Schlegel RA. Back and forth: the regulation and function of transbilayer phospholipid movement in eukaryotic cells. Mol Membr Biol 1994;11:199-216, doi:10.3109/09687689409160430.
  5. Balasubramanian K, Schroit AJ. Aminophospholipid asymmetry: a matter of life and death. Annu Rev Physiol 2003;65:701-34, doi:10.1146/annurev.physiol.65.092101.142459.
  6. Ran S, Downes A, Thorpe PE. Increased exposure of anionic phospholipids on the surface of tumor blood vessels. Cancer Res 2002;62:6132-40.
  7. Ran S, Thorpe PE. Phosphatidylserine is a marker of tumor vasculature and a potential target for cancer imaging and therapy. Int J Radiat Oncol Biol Phys 2002;54:1479-84, doi:10.1016/S0360-3016(02)03928-7.
  8. Huang X, Bennett M, Thorpe PE. A monoclonal antibody that binds anionic phospholipids on tumor blood vessels enhances the antitumor effect of docetaxel on human breast tumors in mice. Cancer Res 2005;65:4408-16, doi:10.1158/0008-5472.CAN-05-0031.
  9. Stace CL, Ktistakis NT. Phosphatidic acid- and phosphatidylserine-binding proteins. Biochim Biophys Acta 2006;1761:913-26.
  10. Burtea C, Laurent S, Lancelot E, et al. Peptidic targeting of phosphatidylserine for the MRI detection of apoptosis in atherosclerotic plaques. Mol Pharm 2009;6:1903-19, doi:10.1021/mp900106m.
  11. Igarashi K, Kaneda M, Yamaji A, et al. A novel phosphatidylserine-binding peptide motif defined by an anti-idiotypic monoclonal antibody. Localization of phosphatidylserine-specific binding sites on protein kinase C and phosphatidylserine decarboxylase. J Biol Chem 1995;270:29075-8, doi:10.1074/jbc.270.49.29075.
  12. Hanshaw RG, Smith BD. New reagents for phosphatidylserine recognition and detection of apoptosis. Bioorg Med Chem 2005;13:5035-42, doi:10.1016/j.bmc.2005.04.071.
  13. Buehler A, Herzog E, Ale A, et al. High resolution tumor targeting in living mice by means of multispectral optoacoustic tomography. EJNMMI Res 2012;2:, doi:10.1186/2191-219X-2-14.
  14. Belhocine TZ, Blankenberg FG. 99mTc-annexin A5 uptake and imaging to monitor chemosensitivity. Methods Mol Med 2005;111:363-80.
  15. Stratton JR, Dewhurst TA, Kasina S, et al. Selective uptake of radiolabeled annexin V on acute porcine left atrial thrombi. Circulation 1995;92:3113-21, doi:10.1161/01.CIR.92.10.3113.
  16. Blankenberg FG, Katsikis PD, Tait JF, et al. Imaging of apoptosis (programmed cell death) with 99mTc annexin V. J Nucl Med 1999;40:184-91.
  17. Beck AW, Luster TA, Miller AF, et al. Combination of a monoclonal anti-phosphatidylserine antibody with gemcitabine strongly inhibits the growth and metastasis of orthotopic pancreatic tumors in mice. Int J Cancer 2006;118:2639-43, doi:10.1002/ijc.21684.
  18. Ran S, He J, Huang X, et al. Antitumor effects of a monoclonal antibody that binds anionic phospholipids on the surface of tumor blood vessels in mice. Clin Cancer Res 2005;11:1551-62, doi:10.1158/1078-0432.CCR-04-1645.
  19. Luster TA, He J, Huang X, et al. Plasma protein beta-2-glycoprotein 1 mediates interaction between the anti-tumor monoclonal antibody 3G4 and anionic phospholipids on endothelial cells. J Biol Chem 2006;281:29863-71, doi:10.1074/jbc.M605252200.
  20. Gerber DE, Stopeck AT, Wong L, et al. Phase I safety and pharmacokinetic study of bavituximab, a chimeric phosphatidylserine-targeting monoclonal antibody, in patients with advanced solid tumors. Clin Cancer Res 2011;17:6888-96, doi:10.1158/1078-0432.CCR-11-1074.
  21. Jennewein M, Lewis MA, Zhao D, et al. Vascular imaging of solid tumors in rats with a radioactive arsenic-labeled antibody that binds exposed phosphatidylserine. Clin Cancer Res 2008;14:1377-85, doi:10.1158/1078-0432.CCR-07-1516.
  22. Yin Y, Kavlie A, Thorpe PE. Fully human anti-phosphatidylserine antibody inhibits the growth of prostate cancer in mice. : ; .
  23. Zhao D, Stafford JH, Zhou H, Thorpe PE. Near-infrared optical imaging of exposed phosphatidylserine in a mouse glioma model. Transl Oncol 2011;4:355-64.
  24. Mandl SJ, Mari C, Edinger M, et al. Multi-modality imaging identifies key times for annexin V imaging as an early predictor of therapeutic outcome. Mol Imaging 2004;3:1-8, doi:10.1162/153535004773861679.
  25. Belhocine T, Steinmetz N, Hustinx R, et al. Increased uptake of the apoptosis-imaging agent (99m)Tc recombinant human annexin V in human tumors after one course of chemotherapy as a predictor of tumor response and patient prognosis. Clin Cancer Res 2002;8:2766-74.
  26. Kown MH, Strauss HW, Blankenberg FG, et al. In vivo imaging of acute cardiac rejection in human patients using (99m)technetium labeled annexin V. Am J Transplant 2001;1:270-7, doi:10.1034/j.1600-6143.2001.001003270.x.
  27. Ishino S, Kuge Y, Takai N, et al. 99mTc-annexin A5 for noninvasive characterization of atherosclerotic lesions: imaging and histological studies in myocardial infarction-prone Watanabe heritable hyperlipidemic rabbits. Eur J Nucl Med Mol Imaging 2007;34:889-99, doi:10.1007/s00259-006-0289-x.
  28. Narula J, Acio ER, Narula N, et al. Annexin-V imaging for noninvasive detection of cardiac allograft rejection. Nat Med 2001;7:1347-52, doi:10.1038/nm1201-1347.
  29. Ingber DE, Madri JA, Folkman J. Endothelial growth factors and extracellular matrix regulate DNA synthesis through modulation of cell and nuclear expansion. In Vitro Cell Dev Biol 1987;23:387-94, doi:10.1007/BF02620997.
  30. He J, Luster TA, Thorpe PE. Radiation-enhanced vascular targeting of human lung cancers in mice with a monoclonal antibody that binds anionic phospholipids. Clin Cancer Res 2007;13:5211-8, doi:10.1158/1078-0432.CCR-07-0793.
  31. He J, Yin Y, Luster TA, et al. Antiphosphatidylserine antibody combined with irradiation damages tumor blood vessels and induces tumor immunity in a rat model of glioblastoma. Clin Cancer Res 2009;15:6871-80, doi:10.1158/1078-0432.CCR-09-1499.
  32. Daleke DL. Regulation of transbilayer plasma membrane phospholipid asymmetry. J Lipid Res 2003;44:233-42, doi:10.1194/jlr.R200019-JLR200.
  33. Zachowski A, Favre E, Cribier S, et al. Outside-inside translocation of aminophospholipids in the human erythrocyte membrane is mediated by a specific enzyme. Biochemistry 1986;25:2585-90, doi:10.1021/bi00357a046.
  34. Stafford JH, Thorpe PE. Increased exposure of phosphatidylethanolamine on the surface of tumor vascular endothelium. Neoplasia 2011;13:299-308.
  35. Blankenberg F. To scan or not to scan, it is a question of timing: technetium-99m-annexin V radionuclide imaging assessment of treatment efficacy after one course of chemotherapy. Clin Cancer Res 2002;8:2757-8.
  36. Amin C, Mackman N, Key NS. Microparticles and cancer. Pathophysiol Haemost Thromb 2007;36:177-83, doi:10.1159/000175155.
  37. Lechner D, Kollars M, Gleiss A, et al. Chemotherapy-induced thrombin generation via procoagulant endothelial microparticles is independent of tissue factor activity. J Thromb Haemost 2007;5:2445-52, doi:10.1111/j.1538-7836.2007.02788.x.
  38. Ke S, Wen X, Wu QP, et al. Imaging taxane-induced tumor apoptosis using PEGylated, 111In-labeled annexin V. J Nucl Med 2004;45:108-15.
  39. Kemerink GJ, Liu X, Kieffer D, et al. Safety, biodistribution, and dosimetry of 99mTc-HYNIC-annexin V, a novel human recombinant annexin V for human application. J Nucl Med 2003;44:947-52.
  40. Lub-de Hooge MN, Kosterink JG, Perik PJ, et al. Preclinical characterisation of 111In-DTPA-trastuzumab. Br J Pharmacol 2004;143:99-106, doi:10.1038/sj.bjp.0705915.
  41. Blankenberg FG, Kalinyak J, Liu L, et al. 99mTc-HYNIC-annexin V SPECT imaging of acute stroke and its response to neuroprotective therapy with anti-Fas ligand antibody. Eur J Nucl Med Mol Imaging 2006;33:566-74, doi:10.1007/s00259-005-0046-6.
  42. Lahorte CM, Vanderheyden JL, Steinmetz N, et al. Apoptosis-detecting radioligands: current state of the art and future perspectives. Eur J Nucl Med Mol Imaging 2004;31:887-919, doi:10.1007/s00259-004-1555-4.
  43. Cauchon N, Langlois R, Rousseau JA, et al. PET imaging of apoptosis with (64)Cu-labeled streptavidin following pretargeting of phosphatidylserine with biotinylated annexin-V. Eur J Nucl Med Mol Imaging 2007;34:247-58, doi:10.1007/s00259-006-0199-y.

Published: May 07, 2013

Views: 253


Send Abstract
Rights & Permissions
Request Reprints