Our goal is to understand the underlying mechanisms regulating lineage specification of hematopoietic stem cells (HSC) and vascular angioblast cells from mesoderm during embryogenesis. As an entry point to achieve this we have chosen to study two molecular factors that are expressed during the early stages of blood and vessel development in zebrafish, ETSRP and DUSP5. The main goal of this proposal is to identify the molecular targets of a novel transcription factor expressed in hemangioblasts, ETSRP, by adopting the newly developed Chromatin Immunoprecipitation-Sequencing (ChlP-Seq) approach, whereby the chromatin regions bound by a transcription factor are immunoprecipitated, amplified and quantified by direct sequencing. Chromatin regions with reads above a predetermined threshold are then computationally mapped to their locations in the genome to identify the targets. This approach is applicable to zebrafish as its genome sequence has recently become available through public databases. Zebrafish is also a superb system for such an application because through subsequent bioinformatics, expression, and functional studies, the potential targets can be validated in vivo and placed in the context of developmental genetic networks. As an example of this validation process, we will examine one genetic entity that is genetically downstream of ETSRP and whose expression pattern is blood and vessel specific, Dual Specific Phosphatase, DUSP5, Both loss and gain of function approaches will be used for this purpose, and it's functional relevance will be examined if it is found to be relevant to circulatory development. These projects bear significant relevance to public health because various clinical disorders such as cancer, rhematoid arthritis, and ischemias either rely on circulatory formation or are caused by problems with circulation. Medicinal approaches to alleviate such disorders depend on careful and methodical research, and basic scientific research such as the work proposed here is focused on providing future therapeutic development with valuable information on the biology of circulation

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31HL091713-05
Application #
8314001
Study Section
Special Emphasis Panel (ZRG1-DIG-E (29))
Program Officer
Meadows, Tawanna
Project Start
2008-09-01
Project End
2013-08-12
Budget Start
2012-09-01
Budget End
2013-08-12
Support Year
5
Fiscal Year
2012
Total Cost
$30,056
Indirect Cost
Name
University of California Los Angeles
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Gomez, Gustavo; Lee, Jae-Hyung; Veldman, Matthew B et al. (2012) Identification of vascular and hematopoietic genes downstream of etsrp by deep sequencing in zebrafish. PLoS One 7:e31658
Ren, Xi; Gomez, Gustavo A; Zhang, Bo et al. (2010) Scl isoforms act downstream of etsrp to specify angioblasts and definitive hematopoietic stem cells. Blood 115:5338-46