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Thromb Haemost 2019; 119(08): 1283-1294
DOI: 10.1055/s-0039-1688907
Coagulation and Fibrinolysis
Georg Thieme Verlag KG Stuttgart · New York

Increased Accumulation and Retention of rhFVIIa (eptacog beta) in Knee Joints of Hemophilia A Mice Compared to Wild-Type Mice

Jhansi Magisetty
1   Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United States
,
Usha R. Pendurthi
1   Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United States
,
SubbaRao V. Madhunapantula
2   Center of Excellence in Molecular Biology and Regenerative Medicine Laboratory, Department of Biochemistry, JSS Medical College, JSS Academy of Higher Education & Research, Mysuru, Karnataka, India
,
Jerry Grandoni 
3   LFB USA, Framingham, Massachusetts, United States
,
L. Vijaya Mohan Rao
1   Department of Cellular and Molecular Biology, The University of Texas Health Science Center at Tyler, Tyler, Texas, United States
› Author Affiliations
Funding This work was supported by grants from HEMA Biologics, LLC and National Heart, Lung, and Blood Institute (HL107483) to L.V.M.R.
Further Information

Publication History

07 September 2018

31 March 2019

Publication Date:
26 May 2019 (online)

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Abstract

Our earlier studies showed that recombinant human factor VIIa (rhFVIIa) administered intravascularly in mice disappeared rapidly from the circulation. However, a small fraction of rhFVIIa that entered extravascular remained functionally active for an extended period. The present study aims to investigate the dose-dependency of rhFVIIa accumulation and retention in mouse knee joints and test whether the hemophilic condition affects rhFVIIa sequestration in joints. Wild-type and FVIII−/− mice were injected with three doses of rhFVIIa (eptacog beta, 90, 250, and 500 μg/kg) via the tail vein. At varying times following rhFVIIa administration, blood and knee joints were collected to measure FVIIa activity and antigen levels in plasma and joint tissues. Joint tissue sections were analyzed by immunohistochemistry for the presence of rhFVIIa. Vascular permeability was assessed by either Evans Blue dye or fluorescein dextran extravasation. The study showed that rhFVIIa accumulated in knee joints of wild-type and FVIII−/− mice in a dose-dependent manner. rhFVIIa antigen and FVIIa activity could be detectable in joints for at least 7 days. Significantly higher levels of rhFVIIa accumulation were observed in knee joints of FVIII−/− mice compared with that of wild-type mice. Immunohistochemical analyses confirmed higher levels of rhFVIIa retention in FVIII−/− mice compared with wild-type mice. Additional studies showed that FVIII−/− mice were more permissible to vascular leakage. In conclusion, the present data demonstrate a dose-dependent accumulation of rhFVIIa in knee joints, and the hemophilic condition enhances the entry of rhFVIIa from circulation to the extravascular. The present data will be useful in improving rhFVIIa prophylaxis.

Keywords

factor VIIa - eptacog beta - hemophilia - permeability - prophylaxis

Authors' Contributions

J.M. performed experiments and analyzed the data. J.G. provided rFVIIa. U.R.P. contributed in designing the study. S.V.M. had an overall supervisory role to J.M. L.V.M.R conceived and designed the research, analyzed the data, and wrote the manuscript. All authors contributed to the preparation of the manuscript.


Supplementary Material

 

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