I, Vladimir Bogdanov, am applying for this research development award in order to finalize my transition into an independent research scientist. The immediate goals are: 1) to acquire new laboratory skills and generate novel research data that would enable me to compete for regular research project grant support (RO1), 2) to acquire first-hand experience in being a principal investigator, and 3) to make substantial progress in elucidating the mechanisms involved in regulation of human tissue factor (TF) pre-mRNA processing in monocytic cells. Specifically, I propose to study regulatory mechanisms responsible for generation of alternatively spliced human tissue factor (asHTF) in monocytes. asHTF, a recently identified molecule, is a soluble, biologically active form of human tissue factor generated by alternative splicing. It is present in circulating blood and thrombi and interacts with platelets under shear conditions. Full-length membrane-bound TF and asHTF are both present in arterial and ex-vivo thrombi. Initial results show that circulating monocytes serve as one of the major sources of asHTF protein in human plasma. Thus, mechanics of tissue factor pre-mRNA splicing in monocytic cells is the primary research objective of the proposed study. In addition, I propose to achieve selective down-regulation of full-length membrane bound TF and asHTF mRNA in monocytic cells by a novel technique of RNA interference and assess the impact of this down-regulation on the total coagulant potential of monocytic cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01DK065752-02
Application #
7015080
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Bishop, Terry Rogers
Project Start
2005-02-07
Project End
2009-01-31
Budget Start
2006-02-01
Budget End
2007-01-31
Support Year
2
Fiscal Year
2006
Total Cost
$113,477
Indirect Cost
Name
Mount Sinai School of Medicine
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
Chandradas, Sajiv; Deikus, Gintaras; Tardos, Jonathan G et al. (2010) Antagonistic roles of four SR proteins in the biosynthesis of alternatively spliced tissue factor transcripts in monocytic cells. J Leukoc Biol 87:147-52
Bogdanov, Vladimir Y; Osterud, Bjarne (2010) Cardiovascular complications of diabetes mellitus: The Tissue Factor perspective. Thromb Res 125:112-8
Eisenreich, Andreas; Bogdanov, Vladimir Y; Zakrzewicz, Andreas et al. (2009) Cdc2-like kinases and DNA topoisomerase I regulate alternative splicing of tissue factor in human endothelial cells. Circ Res 104:589-99
van den Berg, Y W; van den Hengel, L G; Myers, H R et al. (2009) Alternatively spliced tissue factor induces angiogenesis through integrin ligation. Proc Natl Acad Sci U S A 106:19497-502
Bogdanov, V Y; Cimmino, G; Tardos, J G et al. (2009) Assessment of plasma tissue factor activity in patients presenting with coronary artery disease: limitations of a commercial assay. J Thromb Haemost 7:894-7
Tardos, J G; Eisenreich, A; Deikus, G et al. (2008) SR proteins ASF/SF2 and SRp55 participate in tissue factor biosynthesis in human monocytic cells. J Thromb Haemost 6:877-84
Bogdanov, Vladimir Y (2006) Blood coagulation and alternative pre-mRNA splicing: an overview. Curr Mol Med 6:859-69
Bogdanov, V Y; Kirk, R I; Miller, C et al. (2006) Identification and characterization of murine alternatively spliced tissue factor. J Thromb Haemost 4:158-67