In Vitro Platelet Binding Compared with In Vivo Thrombus Imaging Using αIIbβ3-Targeted Radioligands

Linda C. Knight; Jan E. Romano; Alan H. Maurer

doi:10.1055/s-0037-1615369

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1998; 80(05): 845-851
DOI: 10.1055/s-0037-1615369

Review Article

Schattauer GmbH

In Vitro Platelet Binding Compared with In Vivo Thrombus Imaging Using α_IIbβ₃-Targeted Radioligands

Linda C. Knight

¹From the Nuclear Medicine Division, Department of Diagnostic Imaging and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA

,

Jan E. Romano

¹From the Nuclear Medicine Division, Department of Diagnostic Imaging and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA

,

Alan H. Maurer

¹From the Nuclear Medicine Division, Department of Diagnostic Imaging and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA, USA

› Author Affiliations

Abstract

Summary

Radioligands for the αIIbβ3 integrin on platelets are being studied for their ability to image venous thrombi and pulmonary emboli. One such radioligand, ¹²³I-bitistatin, was previously shown to have higher thrombotic uptake in an animal model than other disintegrins, but the reason for this difference was not clear. The purpose of this study was to evaluate three labeled disintegrins, bitistatin, kistrin and barbourin, to look for in vitro differences in platelet binding which could explain the in vivo behavior.

Disintegrins labeled with ¹²⁵I were compared in vitro for extent of binding to platelets and rates of binding and dissociation. These findings were related to organ distribution and image quality for imaging thrombotic lesions, following administration of ¹²³I-disintegrins in an animal model. Fibrinogen at 8.8 μmol/l was able to displace ¹²⁵Ibarbourin and ¹²⁵I-kistrin more rapidly from ADP-stimulated platelets, with half-times of 3.5 and 10.7 min, compared with ¹²⁵I-bitistatin (31.6 min). At equivalent concentrations in whole blood, a higher percentage of bitistatin bound to platelets compared with the other two. In vivo, kistrin and barbourin had significantly lower thrombus:muscle and pulmonary embolus:lung ratios in images compared with bitistatin. There was evidence of more metabolic deiodination of labeled kistrin and barbourin in vivo compared with bitistatin. A surprising finding was that conventional in vitro platelet binding studies did not predict the relative in vivo behavior of labeled disintegrins.

The results suggest that labeled bitistatin has improved targeting of thrombi because it is less easily displaced from stimulated platelets, permitting longer lesion retention. It also appears to have a greater association with resting platelets in the blood, which may increase bioavailability and delay metabolic breakdown.

Full Text

References

References
1 Carter C, Gent M, Leclerc JR. The epidemiology of venous thrombosis. In: Hemostasis and Thrombosis. Basic principles and clinical practice. 2nd ed. Colman RW, Hirsh J, Marder VJ, Salzman EW, eds. Philadelphia: J B Lippincott; 1987: 1185-98.
2 PIOPED.. Value of the ventilation/perfusion scan in acute pulmonary embolism. Results of the prospective investigation of pulmonary embolism diagnosis (PIOPED). JAMA 1990; 263: 2753-9.
3 Hirsh J, Marder VJ, Salzman EW, Hull RD. Treatment of venous thromboembolism. In: Hemostasis and Thrombosis. Basic principles and clinical practice. 2nd ed. Colman RW, Hirsh J, Marder VJ, Salzman EW eds. Philadelphia: J B Lippincott; 1987: 1266-72.
4 Bennett JS, Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine. J Clin Invest 1979; 64: 1393-401.
5 Wagner CL, Mascelli MA, Neblock DS, Weisman HF, Coller BS, Jordan RE. Analysis of GPIIb/IIIa receptor number by quantification of 7E3 binding to human platelets. Blood 1996; 88: 907-14.
6 Coller BS. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of platelet GPIIb/IIIa complex. J Clin Invest 1985; 76: 101-8.
7 Bai Y, Durbin H, Hogg N. Monoclonal antibodies specific for platelet glycoproteins react with human monocytes. Blood 1984; 64: 139-46.
8 Peters AM, Lavender JP, Needham SG, Loutfi I, Snook D, Epenetos AA, Lumley P, Keery RJ, Hogg N. Imaging thrombus with radiolabelled monoclonal antibody to platelets. Brit Med J 1986; 293: 1525-7.
9 Oster ZH, Srivastava SC, Som P, Meinken GE, Scudder LE, Yamamoto K, Atkins HL, Brill AB, Coller BS. Thrombus radioimmunoscintigraphy: An approach using monoclonal antiplatelet antibody. Proc Natl Acad Sci USA 1985; 82: 3465-3468.
10 Knight LC, Radcliffe R, Maurer AH, Rodwell JD, Alvarez VL. Thrombus imaging with technetium-99m synthetic peptides based upon the binding domain of a monoclonal antibody to activated platelets. J Nucl Med 1994; 35: 282-8.
11 Lister-James J, Knight LC, Maurer AH, Bush LR, Moyer BR, Dean RT. Thrombus imaging with a technetium-99m-labeled, activated platelet receptor-binding peptide. J Nucl Med 1996; 37: 775-81.
12 Dennis MS, Henzel WJ, Pitti RM, Lipari MT, Napier MA, Deisher TA, Bunting S, Lazarus RA. Platelet glycoprotein IIb-IIIa protein antagonists from snake venoms: Evidence for a family of platelet-aggregation inhibitors. Proc Natl Acad Sci USA 1990; 87: 2471-5.
13 Gould RJ, Polokoff MA, Friedman PA, Huang T-F, Holt JC, Cook JJ, Niewiarowski S. Disintegrins: A famity of integrin inhibitory proteins from viper venoms. Proc Soc Exp Biol Med 1990; 195: 168-71.
14 Scarborough RM, Rose JW, Hsu MA, Phillips DR, Fried VA, Campbell AM, Nannizzi L, Charo IF. Barbourin. A GPIIb-IIIa-specific integrin antagonist from the venom of Sistrurus m. barbouri . J Biol Chem 1991; 266: 9359-62.
15 Shebuski RJ, Stabilito IJ, Sitko GR, Polokoff MH. Acceleration of recombinant tissue-type plasminogen activator-induced thrombolysis and prevention of reocclusion by the combination of heparin and the Arg-GlyAsp-containing peptide bitistatin in a canine model of coronary thrombosis. Circulation 1990; 82: 169-77.
16 Yasuda T, Gold HK, Leinbach RC, Yaoita H, Fallon JT, Guerrero L, Napier MA, Bunting S, Collen D. Kistrin, a polypeptide platelet GPIIb/IIIa receptor antagonist, enhances and sustains coronary arterial thrombolysis with recombinant tissue-type plasminogen activator in a canine preparation. Circulation 1991; 83: 1038-47.
17 Bernabei A, Gikakis N, Kowalska MA, Niewiarowski S, Edmunds LH. Iloprost and echistatin protect platelets during simulated extracorporeal circulation. Ann Thorac Surg 1995; 59: 149-53.
18 Knight LC, Maurer AH, Romano JE. Comparison of Iodine-123-disinte-grins for imaging thrombi and emboli in a canine model. J Nucl Med 1996; 37: 476-82.
19 Shebuski RJ, Ramjit DR, Bencen GH, Polokoff MA. Characterization and platelet inhibitory activity of bitistatin, a potent arginine-glycine-aspartic acid-containing peptide from the venom of the viper Bitis arietans. J Biol Chem 1989; 264: 21550-6.
20 Fraker PJ, Speck JC. Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3α, 6α-diphenylglycoluril. Biochem Biophys Res Commun 1978; 80: 849-857.
21 Parker RC. Methods of Tissue Culture. 3rd ed. New York: Hoeber Medical Division, Harper and Row; 1962
22 Burt DR. Receptor Binding Methodology and Analysis. In: Receptor Binding in Drug Research. R.A. O’Brien, eds. New York: Marcel Dekker; 1986: 4-29.
23 Knight LC, Maurer AH, Ammar IA, Epps LA, Dean RT, Pak KY, Berger HJ. ^99mTc-Antifibrin Fab’ fragments for imaging venous thrombi: Evaluation in a canine model. Radiology 1989; 173: 163-9.
24 Knight LC. Scintigraphic methods for detecting vascular thrombus. J Nucl Med 1993; 34: 554-61.
25 McLane MA, Kowalska MA, Silver L, Shattil SJ, Niewiarowski S. Interaction of disintegrins with the alpha IIb beta 3 receptor on resting and activated human platelets. Biochemical Journal 1994; 301: 429-36.
26 Scheffel U, Tsan M-F, Mitchell TG, Camargo EE, Braine H, Ezekowitz MD, Nickoloff EL, Hill-Zobel R, McIntyre PA. Human platelets labeled with In-111 8-hydroxyquinoline: Kinetics, distribution, and estimates of radiation dose. J Nucl Med 1982; 23: 149-56.
27 Calvete JJ. Clues for understanding the structure and function of a prototypic human integrin: the platelet glycoprotein IIb/IIIa complex. Thromb Haemost 1994; 72: 1-15.
28 Cerqueira MD, Stratton JR, Vracko R, Schaible TF, Ritchie JL. Noninvasive arterial thrombus imaging with ^99mTc monoclonal antifibrin antibody. Circulation 1992; 85: 298-304.
29 Knight LC, Maurer AH, Ammar IA, Shealy DJ, Mattis JA. Evaluation of ¹¹¹In-labeled anti-fibrin antibody for imaging vascular thrombi. J Nucl Med 1988; 29: 494-502.
30 deFaucal P, Peltier P, Planchon B, Dupas B, Touze M-D, Baron D, Schaible T, Berger HJ, Chatal J-F. Evaluation of 111In-labeled antifibrin monoclonal antibody for the diagnosis of venous thrombotic disease. J Nucl Med 1991; 32: 785-91.
31 Muto P, Lastoria S, Varrella P, Vergara E, Salvatore M, Morgano G, Lister-James J, Bernardy JD, Dean RT, Wencker D, Borer JS. Detecting deep venous thrombosis with technetium-99m-labeled synthetic peptide P280. J Nucl Med 1995; 36: 1384-91.
32 DeNardo SJ, DeNardo GL. Iodine-123 fibrinogen scintigraphy. Semin Nucl Med 1977; 7: 245-51.
33 Becker W, Börner W, Borst U. ^99mTc hexamethylpropyleneamineoxime (HMPAO) as a platelet label: evaluation of labelling parameters and first in vivo results. Nuclear Medicine Communications 1988; 9: 831-42.