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18F-labelled annexin V: a PET tracer for apoptosis imaging

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Abstract

Annexin V can be used to detect apoptotic cells in vitro and in vivo, based on its ability to identify extracellular phosphatidylserine, which arises during apoptosis. In the present study, we examined the synthesis of fluorine-18 labelled annexin V as a positron emission tomography tracer for apoptosis imaging. The distribution of [18F]annexin V and technetium-99m labelled annexin V, a well-characterised SPET tracer for apoptosis imaging, was compared. [18F]annexin V was synthesised using N-succinimidyl 4-[18F]fluorobenzoate as an 18F labelling reagent. Synthesised and purified [18F]annexin V was confirmed by SDS-PAGE. In an ex vivo imaging experiment, [18F]annexin V was intravenously injected into rats 24 h after the induction of myocardial ischaemia, and accumulation in the left ventricle was examined. [18F]annexin V accumulated in the infarct area of the left ventricle, where apoptotic cells were observed. In separate experiments, [18F]annexin V or [99mTc]annexin V was intravenously injected into ischaemic or normal animals, and the distribution of the tracers was compared. In ischaemic animals, accumulation of [18F]annexin V and [99mTc]annexin V in the infarct area was about threefold higher than in the non-infarct area. Furthermore, the ratio of accumulation in the normal heart to the blood radioactivity was not significantly different between the tracers. In normal animals, however, the uptake of [18F]annexin V in the liver, spleen and kidney was much lower than that of [99mTc]annexin V. The low uptake of [18F]annexin V in these organs might represent an advantage over [99mTc]annexin V.

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References

  1. Raynal P, Pollard HB. Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins. Biochim Biophys Acta 1994; 1197: 63–93.

    Article  CAS  PubMed  Google Scholar 

  2. Tait JF, Gibson D, Fujikawa K. Phospholipid binding properties of human placental anticoagulant protein-I, a member of the lipocortin family. J Biol Chem 1989; 264: 7944–7949.

    CAS  PubMed  Google Scholar 

  3. Andree HA, Reutelingsperger CP, Hauptmann R, Hemker HC, Hermens WT, Willems GM. Binding of vascular anticoagulant alpha (VAC alpha) to planar phospholipid bilayers. J Biol Chem 1990; 265:4923–4928.

    CAS  PubMed  Google Scholar 

  4. Thiagarajan P, Tait JF. Binding of annexin V/placental anticoagulant protein I to platelets. Evidence for phosphatidylserine exposure in the procoagulant response of activated platelets. J Biol Chem 1990; 265:17420–17423.

    CAS  PubMed  Google Scholar 

  5. Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol 1992; 148:2207–2216.

    CAS  PubMed  Google Scholar 

  6. Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 1994; 84:1415–1420.

    CAS  PubMed  Google Scholar 

  7. Martin SJ, Reutelingsperger CP, McGahon AJ, et al. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 1995; 182:1545–1556.

    CAS  PubMed  Google Scholar 

  8. Blankenberg FG, Katsikis PD, Tait JF, et al. In vivo detection and imaging of phosphatidylserine expression during programmed cell death. Proc Natl Acad Sci U S A 1998; 95:6349–6354.

    Article  CAS  PubMed  Google Scholar 

  9. Tait JF, Brown DS, Gibson DF, Blankenberg FG, Strauss HW. Development and characterization of annexin V mutants with endogenous chelation sites for (99m)Tc. Bioconjug Chem 2000; 11:918–925.

    Article  CAS  PubMed  Google Scholar 

  10. Blankenberg FG, Katsikis PD, Tait JF, et al. In vivo detection and imaging of phosphatidylserine expression during programmed cell death. Proc Natl Acad Sci U S A 1998; 95:6349–6354.

    Article  CAS  PubMed  Google Scholar 

  11. Blankenberg FG, Katsikis PD, Tait JF, et al. Imaging of apoptosis (programmed cell death) with99mTc annexin V. J Nucl Med 1999; 40:184–191.

    CAS  PubMed  Google Scholar 

  12. Blankenberg FG, Robbins RC, Stoot JH, et al. Radionuclide imaging of acute lung transplant rejection with annexin V. Chest 2000; 117:834–840.

    CAS  PubMed  Google Scholar 

  13. Russell J, O’Donoghue JA, Finn R, et al. Iodination of annexin V for imaging apoptosis. J Nucl Med 2002; 43:671–677.

    Google Scholar 

  14. Vriens PW, Blankenberg FG, Stoot JH, et al. The use of technetium Tc 99m annexin V for in vivo imaging of apoptosis during cardiac allograft rejection. J Thorac Cardiovasc Surg 1998; 116:844–853.

    CAS  PubMed  Google Scholar 

  15. Garg PK, Garg S, Zalutsky MR. Fluorine-18 labeling of monoclonal antibodies and fragments with preservation of immunoreactivity. Bioconjug Chem 1991; 2:44–49.

    CAS  PubMed  Google Scholar 

  16. Guhlke S, Coenen HH, Stocklin G. Fluoroacylation agents based on small n.c.a. [18F]fluorocarboxylic acids. Appl Radiat Isot 1994; 45:715–727.

    CAS  Google Scholar 

  17. Herman LW, Fischman AJ, Tompkins RG, et al. The use of pentafluorophenyl derivatives for the18F labelling of proteins. Nucl Med Biol 1994; 21:1005–1010.

    CAS  PubMed  Google Scholar 

  18. Kilbourn MR, Dence CS, Welch MJ, Mathias CJ. Fluorine-18 labeling of proteins. J Nucl Med 1987; 28:462–470.

    CAS  PubMed  Google Scholar 

  19. Lang L, Eckelman WC. One-step synthesis of18F labeled [18F]-N-succinimidyl 4-(fluoromethyl)benzoate for protein labeling. Appl Radiat Isot 1994; 45:1155–1163.

    Article  CAS  PubMed  Google Scholar 

  20. Shai Y, Kirk KL, Channing MA, et al.18F-labeled insulin: a prosthetic group methodology for incorporation of a positron emitter into peptides and proteins. Biochemistry 1989; 28:4801–4806.

    CAS  PubMed  Google Scholar 

  21. Vaidyanathan G, Zalutsky MR. Labeling proteins with fluorine-18 using N-succinimidyl 4-[18F]fluorobenzoate. Int J Rad Appl Instrum B 1992; 19:275–281.

    CAS  PubMed  Google Scholar 

  22. Wester HJ, Hamacher K, Stocklin G. A comparative study of N.C.A. fluorine-18 labeling of proteins via acylation and photochemical conjugation. Nucl Med Biol 1996; 23:365–372.

    Article  CAS  PubMed  Google Scholar 

  23. Haka MS, Kilbourn MR, Watkins GL, Toorongian SA. Aryltrimethylammonium trifluoromethanesulfonates as precursor to aryl [18F]fluorides: improved synthesis of [18F]GBR-13119. J. Label Compd Radiopharm 1988; 27:823–833.

    Google Scholar 

  24. Kajstura J, Cheng W, Reiss K. Apoptotic and necrotic myocyte cell deaths are independent contributing variables in infarct size in rats. Lab Invest 1996; 74:86–107.

    CAS  PubMed  Google Scholar 

  25. Fliss H, Gattinger D. Apoptosis in ischemic and reperfused rat myocardium. Circ Res 1996; 79:949–956.

    CAS  PubMed  Google Scholar 

  26. Bialik S, Geenen DL, Sasson IE, et al. Myocyte apoptosis during acute myocardial infarction in the mouse localizes to hypoxic regions but occurs independently of p53. J Clin Invest 1997; 100:1363–1372.

    CAS  PubMed  Google Scholar 

  27. Dumont EA, Hofstra L, van Heerde WL, et al. Cardiomyocyte death induced by myocardial ischemia and reperfusion: measurement with recombinant human annexin-V in a mouse model. Circulation 2000; 102:1564–1568.

    CAS  PubMed  Google Scholar 

  28. Hofstra L, Liem IH, Dumont EA, et al. Visualisation of cell death in vivo in patients with acute myocardial infarction. Lancet 2000; 356:209–212.

    Article  CAS  PubMed  Google Scholar 

  29. Albertini A, Zucchini P, Noera G, Cadossi R, Napoleone CP, Pierangeli A. Protective effect of low frequency low energy pulsing electromagnetic fields on acute experimental myocardial infarcts in rats. Bioelectromagnetics 1999; 20:372–377.

    Article  CAS  PubMed  Google Scholar 

  30. Aye NN, Komori S, Hashimoto K. Effects and interaction of cariporide and preconditioning on cardiac arrhythmias and infarction in rat in vivo. Br J Pharmacol 1999; 127:1048–1055.

    CAS  PubMed  Google Scholar 

  31. Bernauer W. The effect of beta-adrenoceptor blocking agents on evolving myocardial necrosis in coronary ligated rats with and without reperfusion. Naunyn Schmiedebergs Arch Pharmacol 1985; 328:288–294.

    CAS  PubMed  Google Scholar 

  32. 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; 8:2766–2774.

    CAS  PubMed  Google Scholar 

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Acknowledgement

The authors thank Toshikazu Ogawa for assistance in the pathophysiological study. The authors also thank Shigeo Hayashi for operation of the cyclotron.

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Correspondence to Yoshihiro Murakami.

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Murakami, Y., Takamatsu, H., Taki, J. et al. 18F-labelled annexin V: a PET tracer for apoptosis imaging. Eur J Nucl Med Mol Imaging 31, 469–474 (2004). https://doi.org/10.1007/s00259-003-1378-8

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  • DOI: https://doi.org/10.1007/s00259-003-1378-8

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