The relationship between thrombospondin 2 (TSP2) and endothelial nitric oxide synthase (eNOS) are not known. Activation of VEGFR-2 via VEGF or by fluid shear stress activates the protein kinase Akt and the endothelial specific Akt substrate, endothelial nitric oxide synthase (eNOS), leading to arteriogenesis and angiogenesis. As inferred from exciting preliminary studies from our laboratories, TSP2 serves as a negative regulator of the Akt-1/eNOS pathway. Previously, we have reported that TSP2 knockout mice (TSP2-K0) demonstrate enhanced arteriogenesis and blood flow recovery in response to limb ischemia and augmented tissue healing. Interestingly, our preliminary studies show that TSP2 levels are elevated in eNOS-KO and Akt1-KO mice, which both display marked impairments in arteriogenesis and angiogenesis and NO, per se, negatively regulates TSP2 levels. Moreover, TSP2/eN0S double knockout (DKO) mice exhibit substantial improvements in blood flow recovery and tissue repair. Thus, we hypothesize that regulation of TSP2 expression by eNOS-derived NO constitutes a previously undefined pro-arteriogenic and pro-angiogenic property of NO. To test this idea, we will: 1. test the hypothesis that the Akt1/eNOS axis mediates arteriogenic and angiogenic responses in vivo, in part, by repressing TSP2 levels;2. determine the mechanism(s) through which the Akt1/eNOS axis and crosstalk with ERK regulates TSP2 and arteriogenesis;and 3. dissect the regulation of the N0/TSP2 pathway and other endothelial cell functions during in vitro angiogenesis. Collectively, these experiments will allow us to delve deeply into the functional antagonism of arteriogenesis and angiogenesis by TSP2, and to delineate how NO regulates TSP2 gene expression and blood flow recovery after limb ischemia.

Public Health Relevance

Peripheral arterial disease is a common disease effecting 10-25% of patients over the age of 55 years old. We propose to investigate the crosstalk between the molecular and biomechanical signals that trigger vascular remodeling and the secretion of regulatory molecules that mediate these effects. Completion of this project should lead to a greater understanding of vascular remodeling and identification of novel therapeutic strategies for arterial disease.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
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Heart, Lung, and Blood Initial Review Group (HLBP)
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Yale University
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Kofler, Natalie; Simons, Michael (2016) The expanding role of neuropilin: regulation of transforming growth factor-β and platelet-derived growth factor signaling in the vasculature. Curr Opin Hematol 23:260-7
Sawyer, Andrew J; Kyriakides, Themis R (2016) Matricellular proteins in drug delivery: Therapeutic targets, active agents, and therapeutic localization. Adv Drug Deliv Rev 97:56-68
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Siragusa, Mauro; Fröhlich, Florian; Park, Eon Joo et al. (2015) Stromal cell-derived factor 2 is critical for Hsp90-dependent eNOS activation. Sci Signal 8:ra81
Bancroft, Tara; Bouaouina, Mohamed; Roberts, Sophia et al. (2015) Up-regulation of thrombospondin-2 in Akt1-null mice contributes to compromised tissue repair due to abnormalities in fibroblast function. J Biol Chem 290:409-22
Eelen, Guy; de Zeeuw, Pauline; Simons, Michael et al. (2015) Endothelial cell metabolism in normal and diseased vasculature. Circ Res 116:1231-44
Simons, Michael; Eichmann, Anne (2015) Molecular controls of arterial morphogenesis. Circ Res 116:1712-24

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