Vascular Endothelial Growth Factor (VEGF)-D Stimulates VEGF-A, Stanniocalcin-1, and Neuropilin-2 and Has Potent Angiogenic Effects

Abstract

Objective-The mature form of human vascular endothelial growth factor-D (hVEGF-D ⌬N⌬C) is an efficient angiogenic factor, but its full mechanism of action has remained unclear. We studied the effects of hVEGF-D ⌬N⌬C in endothelial cells using gene array, signaling, cell culture, and in vivo gene transfer techniques. Methods and Results-Concomitant with the angiogenic and proliferative responses, hVEGF-D⌬N⌬C enhanced the phosphorylation of VEGF receptor-2, Akt, and endothelial nitric oxide synthase. Gene arrays, quantitative reverse transcription-polymerase chain reaction, and Western blot revealed increases in VEGF-A, stanniocalcin-1 (STC1), and neuropilin (NRP) 2 expression by hVEGF-D ⌬N⌬C stimulation, whereas induction with hVEGF-A 165 altered the expression of STC1 and NRP1, another coreceptor for VEGFs. The effects of hVEGF-D ⌬N⌬C were seen only under high-serum conditions, whereas for hVEGF-A 165 , the strongest response was observed under low-serum conditions. The hVEGF-D ⌬N⌬C -induced upregulation of STC1 and NRP2 was also evident in vivo in mouse skeletal muscle treated with hVEGF-D ⌬N⌬C by adenoviral gene delivery. The importance of NRP2 in hVEGF-D ⌬N⌬C signaling was further studied with NRP2 small interfering RNA and NRP antagonist, which were able to block hVEGF-D ⌬N⌬C -induced survival of endothelial cells. Key Words: angiogenesis Ⅲ endothelium Ⅲ gene expression Ⅲ growth factors Ⅲ vascular biology T herapeutic angiogenesis is a potential tool for the treatment of cardiovascular diseases. Delivery of angiogenic factors can be used to stimulate the formation of new capillaries and collateral vessels in ischemic myocardium and lower limbs. By this method, it may be possible to increase blood perfusion in ischemic muscles, especially in those patients for whom conventional revascularization strategies, such as angioplasty and bypass surgery, are not suitable. 1,2 Vascular endothelial growth factors (VEGFs) are effective compounds for therapeutic angiogenesis. They are dimeric glycoproteins that regulate endothelial cell behavior, angiogenesis, and lymphangiogenesis. The VEGF family contains at least 6 members: VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, and placental growth factor. Their signaling is mediated primarily via 3 tyrosine kinase receptors: VEGF receptor (VEGFR)-1 (Flt-1), VEGFR-2 (Flk-1/KDR), and VEGFR-3 (Flt-4). In addition, 2 coreceptors for VEGFs, neuropilin (NRP)-1 and NRP2, have been identified. [1][2][3][4] A novel member of the VEGF family, VEGF-D, was initially described as a c-fos-induced growth factor. 5,6 It is synthesized as a large precursor protein that is proteolytically processed by plasmin and some other serine proteases to release the C-and N-terminal peptides. The precursor form of VEGF-D binds to and preferentially activates VEGFR-3, but after proteolytic cleavage, the binding affinity to the VEGFR-2 is increased many fold. 6 -9 Unlike VEGF-A, VEGF-D is not regulated by hypoxia, but regulation by cell-to-cell contacts mediated by cadherin-11 has been reported in fibroblasts. ...

Synergistic effect of p53 on TSA-induced stanniocalcin 1 expression in human nasopharyngeal carcinoma cells, CNE2

“…A role of STC1 in the tumor microenvironment has also been suggested (Yeung et al 2012). This presumption is strengthened by the involvement of STC1 in local inflammation (Sheikh--Hamad 2010) as well as the identification of STC1 as a hypoxia and angiogenic responsive gene (Yeung et al 2005, Law et al 2010, Jauhiainen et al 2011, Roch & Sherwood 2011. To date, biological roles of STC1 in carcinogenesis have been suggested, including cell motility, invasion, and apoptosis (Lai et al 2007, Law et al 2008, Liu et al 2010.…”

Nanoparticle-Mediated Delivery of Pioglitazone Enhances Therapeutic Neovascularization in a Murine Model of Hindlimb Ischemia

“…20 Blood flow recovery by Pio-NP was accompanied by arteriogenesis in ischemic hindlimb muscle, which is demonstrated as arterioles with α-smooth muscle actin-positive pericytes. Polymerase chain reaction array revealed that Pio-NP induced multiple genes that are involved in endothelial proliferation and migration (VEGF-A, VEGF-B, VEGF-D, and epidermal growth factor), [21][22][23][24][25][26] as well as vascular smooth muscle cell migration and proliferation (FGF and Gna13). [27][28][29][30] This coordinated induction of endothelial cells and VSMCs is critical for Pio-NP-induced mature arteriogenesis to form functional collaterals.…”

Semaphorin 3B Gene Suppresses Tumor Growth Through the p53 Signaling Pathway and Neuropilin Receptors

“…It has been shown that SEMA3B protein competitively binds with NP-1 and inhibits VEGFl65-induced tumor growth by inhibiting tumor angiogenesis and reducing the microvessel density (MVD) [22]. A study also found that the expression of SEMA3B mRNA in lung cancer tissue was negatively correlated with the MVD and the expression of VEGF mRNA [23].…”