The major scientific goal of this Pathway to Independence (K99/ROO) Career Development Award application is to understand the molecular pathophysiology of thrombotic and inflammatory disorders by studying novel mechanisms for cytoprotective actions of vitamin K-dependent coagulation proteases. This application focuses initially on the role of membrane receptors in the regulation of the cellular protein C pathway and later on the exploration of novel mechanisms for cytoprotective activities of coagulation proteases. The major career development goal of the applicant is to expand his technical and academic experience required for a successful transition into an independent investigator. These studies will provide the opportunity and solid basis to apply successfully for future independent NIH R01 funding focused on the molecular mechanistic studies centered on the crossroads of coagulation and inflammation. Novel hypotheses on the functional proteomics of cytoprotective actions by blood coagulation proteases will be tested using biochemical and cellular biology methods. The clinical and therapeutic implications of the proposed studies are clear from the large clinical trials, where activated protein C (ARC), but not other anticoagulants reduced mortality in severe sepsis patients and implied that the unique combination of APC's anticoagulant activity and direct activity on cells is the basis for APC's success. My published work and unpublished preliminary data lead directly to the proposed studies and provide strong support for my hypotheses. In testing these hypotheses, I propose: 1) To characterize the formation of endothelial cell membrane receptor complexes between thrombomodulin, endothelial protein C receptor and protease activated receptor-1 required for APC generation and APC's direct effects on cells; 2) To clarify the potential beneficial and detrimental functional properties of platelet factor 4 for APC generation and APC's direct effects on cells; 3) To identify novel themes and mechanisms for APC and fVlla cytoprotective actions on cells by exploration of the similarities and differences between APC and fVlla anti-apoptotic activities; and 4) To establish whether meizothrombin has anti-apoptotic activity, as predicted, and if this activity requires cofactor-dependent and PAR-dependent mechanisms. If the proposed studies are successful, they will increase our knowledge and may lead to improved treatment of a variety of disorders in which thrombosis, apoptosis and inflammation contribute to pathogenesis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Transition Award (R00)
Project #
4R00HL087618-02
Application #
7534741
Study Section
Special Emphasis Panel (NSS)
Program Officer
Sarkar, Rita
Project Start
2006-12-01
Project End
2011-01-31
Budget Start
2008-02-01
Budget End
2009-01-31
Support Year
2
Fiscal Year
2008
Total Cost
$248,999
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Wyseure, Tine; Mosnier, Laurent O; von Drygalski, Annette (2016) Advances and challenges in hemophilic arthropathy. Semin Hematol 53:10-9
Burnier, Laurent; Mosnier, Laurent O (2013) Novel mechanisms for activated protein C cytoprotective activities involving noncanonical activation of protease-activated receptor 3. Blood 122:807-16
von Drygalski, Annette; Furlan-Freguia, Christian; Ruf, Wolfram et al. (2013) Organ-specific protection against lipopolysaccharide-induced vascular leak is dependent on the endothelial protein C receptor. Arterioscler Thromb Vasc Biol 33:769-76
Mosnier, Laurent O; Sinha, Ranjeet K; Burnier, Laurent et al. (2012) Biased agonism of protease-activated receptor 1 by activated protein C caused by noncanonical cleavage at Arg46. Blood 120:5237-46
Mosnier, Laurent O (2011) Platelet factor 4 inhibits thrombomodulin-dependent activation of thrombin-activatable fibrinolysis inhibitor (TAFI) by thrombin. J Biol Chem 286:502-10
Niessen, Frank; Furlan-Freguia, Christian; Fernández, José A et al. (2009) Endogenous EPCR/aPC-PAR1 signaling prevents inflammation-induced vascular leakage and lethality. Blood 113:2859-66
Mosnier, Laurent O; Zampolli, Antonella; Kerschen, Edward J et al. (2009) Hyperantithrombotic, noncytoprotective Glu149Ala-activated protein C mutant. Blood 113:5970-8
Yang, Xia V; Banerjee, Yajnavalka; Fernandez, Jose A et al. (2009) Activated protein C ligation of ApoER2 (LRP8) causes Dab1-dependent signaling in U937 cells. Proc Natl Acad Sci U S A 106:274-9