Characterization of high molecular weight plasma protein complexes induced by clotting factor rFXIII‐treatment in the cynomolgus monkey

Characterization of high molecular weight plasma protein complexes induced by clotting factor rFXIII‐treatment in the cynomolgus monkey

Background: In cynomolgus monkeys, suprapharmacological doses of clotting recombinant factor XIII (rFXIII) cause a generalized coagulopathy, associated with formation of circulating high molecular weight protein complexes (HMEX). HMEX consist of plasma protein substrates cross‐linked by FXIII transg...

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Bibliographic Details
Published in:Journal of thrombosis and haemostasis Vol. 5; no. 10; pp. 2070 - 2078
Main Authors: SCHAAL‐JENSEN, R., KIEHR, B., BOESEN, H. T., KRABBE, J. S., SOMMER, C., JACOBSEN, H., OLEKSIEWICZ, M. B.
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Publishing Ltd 01-10-2007
Subjects:
Animals
biomarker
Blood Coagulation
Blood Coagulation Factors - chemistry
Blood Proteins - chemistry
cynomolgus monkey
Dose-Response Relationship, Drug
Factor XIII - chemistry
Factor XIII - pharmacology
Female
high molecular weight plasma protein complexes
Humans
Kinetics
Macaca fascicularis
Male
Models, Biological
Molecular Weight
recombinant factor XIII
Recombinant Proteins - chemistry
toxicity
Western blot
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Summary:Background: In cynomolgus monkeys, suprapharmacological doses of clotting recombinant factor XIII (rFXIII) cause a generalized coagulopathy, associated with formation of circulating high molecular weight protein complexes (HMEX). HMEX consist of plasma protein substrates cross‐linked by FXIII transglutaminase activity. Objective: To characterize HMEX, with a view to develop safety biomarker assays. Methods: Cynomolgus monkeys received single i.v. injections with vehicle or rFXIII at 1, 3 and 10 mg kg−1. Plasma HMEX were analyzed by sodium dodecylsulfate–polyacrylmide gel electrophoresis, silver staining, Western blotting and quantitative dissociation‐enhanced lanthanide fluoroimmunoassay. Plasma FXIII antigen was analyzed by quantitative ELISA. Human HMEX were made in vitro, by spiking plasma with thrombin‐activated rFXIII. Results: Maximal circulating HMEX levels were reached within 1 h of rFXIII treatment, and remained stable over 24 h. HMEX above 250 kDa contained fibrinogen α‐chains and fibronectin. Fibrinogen γ‐chain was detected only in HMEX below 250 kDa. The total plasma concentration of HMEX was in the low μg mL−1 range, distributed on less than 20 main species. Human and cynomolgus HMEX were similar. HMEX formation increased with rFXIII dose in a disproportionate manner, with 3‐fold and fortyfold increases in HMEX exposure associated with rFXIII dose increments from 1 to 3 and 3 to 10 mg kg−1, respectively. Conclusions: The disproportionate HMEX formation parallels the steep toxicity dose response previously reported for rFXIII in cynomolgus monkeys, supporting a mechanistical role for HMEX in the generalized coagulopathy seen in rFXIII toxicity. Our findings support that HMEX constitute candidate (potential) safety biomarkers in rFXIII treatment.
Bibliography:These authors contributed equally to this work.
ISSN:1538-7933
1538-7836
1538-7836
DOI:10.1111/j.1538-7836.2007.02695.x