In this new proposal, we investigate our novel finding that a unique combination of hematopoietic stem cell cytokines (i.e. stem cell factor-SCF;interleukin 3- IL-3;and stromal-derived factor-1 alpha- SDF-1?) control human endothelial cell (EC) tube morphogenesis and EC-pericyte tube coassembly under defined serum-free conditions (and in the absence of phorbol ester) in 3D collagen matrices. Interestingly, VEGF has no influence by itself and requires the synergistic action of these hematopoietic factors in order to exert an effect on EC sprouting, its major morphogenic influence. We propose the hypothesis that SCF, IL-3 and SDF-1? are critical regulators of vascular morphogenesis which act in a synergistic manner and are necessary cofactors for the action of known mediators of developmental vascularization (e.g. VEGF and BMP-4). Combined administration of c-Kit (the SCF receptor) and CXCR4 (the SDF-1? receptor) inhibitors or blocking antibodies to SCF and IL-3 leads to vascular hemorrhage phenotypes in developing quail embryos that relates to defects in vessel formation and remodeling. Also, we show that the three factors synergize to activate Src, B-Raf and Erk kinases and induce the expression of the ?2?1 integrin and Pak2 which are known regulators of vascular morphogenesis. We will elucidate the molecular basis for the signaling synergism of the three cytokines in ECs, how these factors interface with VEGF, BMP-4, and pericytes to control EC sprouting and how they affect both ECs and pericytes during tube coassembly and maturation events in vitro and in vivo. We propose three specific aims which are;
Specific Aim #1. To elucidate the signaling mechanisms by which SCF, IL-3 and SDF-1? synergize to stimulate vasculogenic EC tube assembly in 3D extracellular matrices.
Specific Aim #2. To determine how SCF, IL-3 and SDF-1? affect VEGF and BMP-4 signaling to control EC sprouting responses in 3D extracellular matrices.
Specific Aim #3. To determine how SCF, IL-3 and SDF-1? act to regulate pericyte-induced EC sprouting responses and vasculogenic EC-pericyte tube coassembly in 3D extracellular matrices.

Public Health Relevance

This work focuses on the ability of factors which are known to support the growth of immature blood cells to also be able to support the formation of blood vessels which carry blood cells. These factors bind to endothelial cells, which line blood vessels and to pericytes, a supporting cell that helps blood vessels form and become stable. A basic understanding of the mechanisms underlying how blood vessels form and become stabilized is critical in efforts to stimulate or inhibit the process in the context of various human diseases such as cardiovascular disease, diabetes or cancer.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL105606-02
Application #
8207865
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Gao, Yunling
Project Start
2011-01-03
Project End
2014-12-31
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
2
Fiscal Year
2012
Total Cost
$378,750
Indirect Cost
$128,750
Name
University of Missouri-Columbia
Department
Pharmacology
Type
Schools of Medicine
DUNS #
153890272
City
Columbia
State
MO
Country
United States
Zip Code
65211
Kim, Dae Joong; Martinez-Lemus, Luis A; Davis, George E (2013) EB1, p150Glued, and Clasp1 control endothelial tubulogenesis through microtubule assembly, acetylation, and apical polarization. Blood 121:3521-30
Davis, George E; Kim, Dae Joong; Meng, Chun-Xia et al. (2013) Control of vascular tube morphogenesis and maturation in 3D extracellular matrices by endothelial cells and pericytes. Methods Mol Biol 1066:17-28
Morin, Kristen T; Smith, Annie O; Davis, George E et al. (2013) Aligned human microvessels formed in 3D fibrin gel by constraint of gel contraction. Microvasc Res 90:12-22
Stratman, Amber N; Davis, George E (2012) Endothelial cell-pericyte interactions stimulate basement membrane matrix assembly: influence on vascular tube remodeling, maturation, and stabilization. Microsc Microanal 18:68-80
Sacharidou, Anastasia; Stratman, Amber N; Davis, George E (2012) Molecular mechanisms controlling vascular lumen formation in three-dimensional extracellular matrices. Cells Tissues Organs 195:122-43
Davis, George E; Stratman, Amber N; Sacharidou, Anastasia et al. (2011) Molecular basis for endothelial lumen formation and tubulogenesis during vasculogenesis and angiogenic sprouting. Int Rev Cell Mol Biol 288:101-65
Stratman, Amber N; Davis, Michael J; Davis, George E (2011) VEGF and FGF prime vascular tube morphogenesis and sprouting directed by hematopoietic stem cell cytokines. Blood 117:3709-19