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Open Access 01-12-2011 | Original research

Failure of annexin-based apoptosis imaging in the assessment of antiangiogenic therapy effects

Authors: Wiltrud Lederle, Susanne Arns, Anne Rix, Felix Gremse, Dennis Doleschel, Jörn Schmaljohann, Felix M Mottaghy, Fabian Kiessling, Moritz Palmowski

Published in: EJNMMI Research | Issue 1/2011

Abstract

Background

Molecular apoptosis imaging is frequently discussed to be useful for monitoring cancer therapy. We demonstrate that the sole assessment of therapy effects by apoptosis imaging can be misleading, depending on the therapy effect on the tumor vasculature.

Methods

Apoptosis was investigated by determining the uptake of Annexin Vivo by optical imaging (study part I) and of ^{99 m}Tc-6-hydrazinonicotinic [HYNIC]-radiolabeled Annexin V by gamma counting (study part II) in subcutaneous epidermoid carcinoma xenografts (A431) in nude mice after antiangiogenic treatment (SU11248). Optical imaging was performed by optical tomography (3D) and 2D reflectance imaging (control, n = 7; therapy, n = 6). Accumulation of the radioactive tracer was determined ex vivo (control, n = 5; therapy, n = 6). Tumor vascularization was investigated with an optical blood pool marker (study part I) and contrast-enhanced ultrasound (both studies). Data were validated by immunohistology.

Results

A significantly higher apoptosis rate was detected in treated tumors by immunohistological terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining (area fraction: control, 0.023 ± 0.015%; therapy, 0.387 ± 0.105%; P < 0.001). However, both 2D reflectance imaging using Annexin Vivo (control, 13 ± 15 FI/cm²; therapy, 11 ± 7 FI/cm²) and gamma counting using ^{99 m}Tc-HYNIC-Annexin V (tumor-to-muscle ratio control, 5.66 ± 1.46; therapy, 6.09 ± 1.40) failed in showing higher accumulation in treated tumors. Optical tomography even indicated higher probe accumulation in controls (control, 81.3 ± 73.7 pmol/cm³; therapy, 27.5 ± 34.7 pmol/cm³). Vascularization was strongly reduced after therapy, demonstrated by contrast-enhanced ultrasound, optical imaging, and immunohistology.

Conclusions

The failure of annexin-based apoptosis assessment in vivo can be explained by the significant breakdown of the vasculature after therapy, resulting in reduced probe/tracer delivery. This favors annexin-based apoptosis imaging only in therapies that do not severely interfere with the vasculature.

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Lahorte CM, Vanderheyden JL, Steinmetz N, Van de WC, Dierckx RA, Slegers G: Apoptosis-detecting radioligands: current state of the art and future perspectives. Eur J Nucl Med Mol Imaging 2004, 31: 887–919. 10.1007/s00259-004-1555-4PubMedCrossRef

Blankenberg FG: In vivo detection of apoptosis. J Nucl Med 2008,49(Suppl 2):81S-95S.PubMedCrossRef

Blankenberg FG, Katsikis PD, Tait JF, Davis RE, Naumovski L, Ohtsuki K, Kopiwoda S, Abrams MJ, Drakes M, Robbins RC, Maecker HT, Strauss HW: In vivo detection and imaging of phosphatidylserine expression during programmed cell death. Proc Natl Acad Sci USA 1998, 95: 6349–6354. 10.1073/pnas.95.11.6349PubMedCentralPubMedCrossRef

De Saint-Hubert M, Prinsen K, Mortelmans L, Verbruggen A, Mottaghy FM: Molecular imaging of cell death. Methods 2009, 48: 178–187. 10.1016/j.ymeth.2009.03.022PubMedCrossRef

De Saint-Hubert M, Mottaghy FM, Vunckx K, Nuyts J, Fonge H, Prinsen K, Stroobants S, Mortelmans L, Deckers N, Hofstra L, Reutelingsperger CP, Verbruggen A, Rattat D: Site-specific labeling of 'second generation' annexin V with 99 mTc(CO)3 for improved imaging of apoptosis in vivo. Bioorg Med Chem 2010, 18: 1356–1363. 10.1016/j.bmc.2009.12.021PubMedCrossRef

Bauwens M, De Saint-Hubert M, Devos E, Deckers N, Reutelingsperger C, Mortelmans L, Himmelreich U, Mottaghy FM, Verbruggen A: Site-specific 68Ga-labeled Annexin A5 as a PET imaging agent for apoptosis. Nucl Med Biol 2011, 38: 381–392. 10.1016/j.nucmedbio.2010.09.008PubMedCrossRef

Petrovsky A, Schellenberger E, Josephson L, Weissleder R, Bogdanov A Jr: Near-infrared fluorescent imaging of tumor apoptosis. Cancer Res 2003, 63: 1936–1942.PubMed

Schellenberger EA, Bogdanov A Jr, Hogemann D, Tait J, Weissleder R, Josephson L: Annexin V-CLIO: a nanoparticle for detecting apoptosis by MRI. Mol Imaging 2002, 1: 102–107. 10.1162/153535002320162769PubMedCrossRef

Krishnan AS, Neves AA, de Backer MM, Hu DE, Davletov B, Kettunen MI, Brindle KM: Detection of cell death in tumors by using MR imaging and a gadolinium-based targeted contrast agent. Radiology 2008, 246: 854–862. 10.1148/radiol.2463070471PubMedCrossRef

10.

Narula J, Acio ER, Narula N, Samuels LE, Fyfe B, Wood D, Fitzpatrick JM, Raghunath PN, Tomaszewski JE, Kelly C, Steinmetz N, Green A, Tait JF, Leppo J, Blankenberg FG, Jain D, Strauss HW: Annexin-V imaging for noninvasive detection of cardiac allograft rejection. Nat Med 2001, 7: 1347–1352. 10.1038/nm1201-1347PubMedCrossRef

11.

Belhocine T, Steinmetz N, Hustinx R, Bartsch P, Jerusalem G, Seidel L, Rigo P, Green A: Increased uptake of the apoptosis-imaging agent (99 m)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: 2766–2774.PubMed

12.

Schellenberger EA, Bogdanov A Jr, Petrovsky A, Ntziachristos V, Weissleder R, Josephson L: Optical imaging of apoptosis as a biomarker of tumor response to chemotherapy. Neoplasia 2003, 5: 187–192.PubMedCentralPubMedCrossRef

13.

Choi HK, Yessayan D, Choi HJ, Schellenberger E, Bogdanov A, Josephson L, Weissleder R, Ntziachristos V: Quantitative analysis of chemotherapeutic effects in tumors using in vivo staining and correlative histology. Cell Oncol 2005, 27: 183–190.PubMed

14.

Manning HC, Merchant NB, Foutch AC, Virostko JM, Wyatt SK, Shah C, McKinley ET, Xie J, Mutic NJ, Washington MK, LaFleur B, Tantawy MN, Peterson TE, Ansari MS, Baldwin RM, Rothenberg ML, Bornhop DJ, Gore JC, Coffey RJ: Molecular imaging of therapeutic response to epidermal growth factor receptor blockade in colorectal cancer. Clin Cancer Res 2008, 14: 7413–7422. 10.1158/1078-0432.CCR-08-0239PubMedCentralPubMedCrossRef

15.

Palmowski M, Huppert J, Hauff P, Reinhardt M, Schreiner K, Socher MA, Hallscheidt P, Kauffmann GW, Semmler W, Kiessling F: Vessel fractions in tumor xenografts depicted by flow- or contrast-sensitive three-dimensional high-frequency Doppler ultrasound respond differently to antiangiogenic treatment. Cancer Res 2008, 68: 7042–7049. 10.1158/0008-5472.CAN-08-0285PubMedCrossRef

16.

Mendel DB, Laird AD, Xin X, Louie SG, Christensen JG, Li G, Schreck RE, Abrams TJ, Ngai TJ, Lee LB, Murray LJ, Carver J, Chan E, Moss KG, Haznedar JO, Sukbuntherng J, Blake RA, Sun L, Tang C, Miller T, Shirazian S, McMahon G, Cherrington JM: In vivo antitumor activity of SU11248, a novel tyrosine kinase inhibitor targeting vascular endothelial growth factor and platelet-derived growth factor receptors: determination of a pharmacokinetic/pharmacodynamic relationship. Clin Cancer Res 2003, 9: 327–337.PubMed

17.

Zhang C, Jugold M, Woenne EC, Lammers T, Morgenstern B, Mueller MM, Zentgraf H, Bock M, Eisenhut M, Semmler W, Kiessling F: Specific targeting of tumor angiogenesis by RGD-conjugated ultrasmall superparamagnetic iron oxide particles using a clinical 1.5-T magnetic resonance scanner. Cancer Res 2007, 67: 1555–1562. 10.1158/0008-5472.CAN-06-1668PubMedCrossRef

18.

Palmowski M, Lederle W, Gaetjens J, Socher M, Hauff P, Bzyl J, Semmler W, Gunther RW, Kiessling F: Comparison of conventional time-intensity curves vs. maximum intensity over time for post-processing of dynamic contrast-enhanced ultrasound. Eur J Radiol 2010, 75: e149-e153. 10.1016/j.ejrad.2009.10.030PubMedCrossRef

19.

Rix A, Lederle W, Siepmann M, Fokong S, Behrendt FF, Bzyl J, Grouls C, Kiessling F, Palmowski M: Evaluation of high frequency ultrasound methods and contrast agents for characterising tumor response to anti-angiogenic treatment. Eur J Radiol, in press.

20.

Montet X, Ntziachristos V, Grimm J, Weissleder R: Tomographic fluorescence mapping of tumor targets. Cancer Res 2005, 65: 6330–6336. 10.1158/0008-5472.CAN-05-0382PubMedCrossRef

21.

Lederle W, Linde N, Heusel J, Bzyl J, Woenne EC, Zwick S, Skobe M, Kiessling F, Fusenig NE, Mueller MM: Platelet-derived growth factor-B normalizes micromorphology and vessel function in vascular endothelial growth factor-A-induced squamous cell carcinomas. Am J Pathol 2010, 176: 981–994. 10.2353/ajpath.2010.080998PubMedCentralPubMedCrossRef

22.

Reshef A, Shirvan A, kselrod-Ballin A, Wall A, Ziv I: Small-molecule biomarkers for clinical PET imaging of apoptosis. J Nucl Med 2010, 51: 837–840. 10.2967/jnumed.109.063917PubMedCrossRef

23.

Wolters SL, Corsten MF, Reutelingsperger CP, Narula J, Hofstra L: Cardiovascular molecular imaging of apoptosis. Eur J Nucl Med Mol Imaging 2007, 34: S86-S98. 10.1007/s00259-007-0443-0PubMedCrossRef

24.

Neves AA, Brindle KM: Assessing responses to cancer therapy using molecular imaging. Biochim Biophys Acta 2006, 1766: 242–261.PubMed

25.

Hoebers FJ, Kartachova M, de BJ, van den Brekel MW, van TH, van HM, Rasch CR, Valdés Olmos RA, Verheij M: 99 mTc Hynic-rh-Annexin V scintigraphy for in vivo imaging of apoptosis in patients with head and neck cancer treated with chemoradiotherapy. Eur J Nucl Med Mol Imaging 2008, 35: 509–518. 10.1007/s00259-007-0624-xPubMedCentralPubMedCrossRef

26.

Marzola P, Degrassi A, Calderan L, Farace P, Nicolato E, Crescimanno C, Sandri M, Giusti A, Pesenti E, Terron A, Sbarbati A, Osculati F: Early antiangiogenic activity of SU11248 evaluated in vivo by dynamic contrast-enhanced magnetic resonance imaging in an experimental model of colon carcinoma. Clin Cancer Res 2005, 11: 5827–5832. 10.1158/1078-0432.CCR-04-2655PubMedCrossRef

27.

Marzola P, Degrassi A, Calderan L, Farace P, Crescimanno C, Nicolato E, Giusti A, Pesenti E, Terron A, Sbarbati A, Abrams T, Murray L, Osculati F: In vivo assessment of antiangiogenic activity of SU6668 in an experimental colon carcinoma model. Clin Cancer Res 2004, 10: 739–750. 10.1158/1078-0432.CCR-0828-03PubMedCrossRef

Title: Failure of annexin-based apoptosis imaging in the assessment of antiangiogenic therapy effects
Authors: Wiltrud Lederle
Susanne Arns
Anne Rix
Felix Gremse
Dennis Doleschel
Jörn Schmaljohann
Felix M Mottaghy
Fabian Kiessling
Moritz Palmowski
Publication date: 01-12-2011
Publisher: Springer Berlin Heidelberg
Published in: EJNMMI Research / Issue 1/2011
Electronic ISSN: 2191-219X
DOI: https://doi.org/10.1186/2191-219X-1-26

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Failure of annexin-based apoptosis imaging in the assessment of antiangiogenic therapy effects

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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