Fibrinogen is a multifunctional plasma protein that after conversion into fibrin contributes to hemostasis and other physiological and pathological processes through its interaction with different proteins and cell types. Among these processes is inflammation, which plays a pivotal role in the pathophysiology of cardiovascular diseases. Recruitment of leukocytes from the circulation to sites of inflammation is an integral part of the inflammatory response and transendothelial migration of leukocytes is a key step in such recruitment. Numerous data indicate that fibrinogen is involved in this process. According to the proposed hypotheses, fibrinogen or its degradation products induce leukocyte transmigration by bridging leukocytes to the endothelium through the interaction with the endothelial receptors ICAM-1 or VE-cadherin, respectively. We have recently discovered that fibrin ?N-domains interact with the VLDL receptor (VLDLR), another receptor on endothelial cells, and this interaction also promotes leukocyte transmigration. Based on this discovery and some preliminary data, we hypothesize that fibrin promotes transendothelial migration of leukocytes through its interaction with VLDLR and such fibrin-induced VLDLR-dependent leukocyte transmigration can be modulated by specific inhibitors targeting this interaction. The major goal of the present application is to test this hypothesis. This will be accomplished in the following specific aims.
The first aim i s to further prove that fibrin-VLDLR interaction promotes leukocyte transmigration by studying the effect of fibrin on this process.
The second aim i s to establish the molecular mechanism of this interaction by mapping the complementary binding sites using recombinant techniques and characterizing the interaction between them using biochemical and biophysical methods.
The third aim i s to develop novel specific inhibitors of this interaction based on knowledge obtained and test their anti-inflammatory properties and cardioprotective effect using in vivo mouse models. The proposed study will clarify the molecular mechanisms of fibrin-dependent inflammation and may result in novel therapeutics for treatment of inflammation-related cardiovascular diseases including myocardial ischemia-reperfusion injury.

Public Health Relevance

Inflammatory mechanisms play a pivotal role in the pathophysiology of cardiovascular diseases. They involve various factors including fibrin(ogen), which promotes transendothelial migration of leukocytes and thereby inflammation. The proposed studies will establish the molecular mechanism of the recently discovered VLDL receptor-fibrin interaction that modulates fibrin-dependent leukocyte transmigration, identify novel inhibitors of this process, and test their anti-inflammatory properties and cardioprotective effect using animal models.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL056051-14A1
Application #
8578983
Study Section
Hemostasis and Thrombosis Study Section (HT)
Program Officer
Link, Rebecca P
Project Start
1998-01-01
Project End
2017-05-31
Budget Start
2013-08-10
Budget End
2014-05-31
Support Year
14
Fiscal Year
2013
Total Cost
$376,288
Indirect Cost
$131,149
Name
University of Maryland Baltimore
Department
Biochemistry
Type
Schools of Medicine
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
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Riedel, Tomas; Suttnar, Jiri; Brynda, Eduard et al. (2011) Fibrinopeptides A and B release in the process of surface fibrin formation. Blood 117:1700-6
Tsurupa, Galina; Mahid, Ariza; Veklich, Yuri et al. (2011) Structure, stability, and interaction of fibrin ?C-domain polymers. Biochemistry 50:8028-37
Yakovlev, S; Gao, Y; Cao, C et al. (2011) Interaction of fibrin with VE-cadherin and anti-inflammatory effect of fibrin-derived fragments. J Thromb Haemost 9:1847-55
Koo, J; Rafailovich, M H; Medved, L et al. (2010) Evaluation of fibrinogen self-assembly: role of its ýýC region. J Thromb Haemost 8:2727-35
Tsurupa, Galina; Yakovlev, Sergiy; McKee, Patrick et al. (2010) Noncovalent interaction of alpha(2)-antiplasmin with fibrin(ogen): localization of alpha(2)-antiplasmin-binding sites. Biochemistry 49:7643-51
Tsurupa, Galina; Hantgan, Roy R; Burton, Robert A et al. (2009) Structure, stability, and interaction of the fibrin(ogen) alphaC-domains. Biochemistry 48:12191-201
Yakovlev, Sergiy; Medved, Leonid (2009) Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: localization of the fibrin-binding site within the third extracellular VE-cadherin domain. Biochemistry 48:5171-9
Medved, L; Weisel, J W; Fibrinogen and Factor XIII Subcommittee of Scientific Standardization Committee of International Society on Thrombosis and Haemostasis (2009) Recommendations for nomenclature on fibrinogen and fibrin. J Thromb Haemost 7:355-9

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