Down Syndrome Candidate Region 1,a Downstream Target of VEGF, Participa tes in Endothelial Cell Migration and Angiogenesis

Abstract

Vascular endothelial growth factor (VEGF) is a principal stimulator of angiogenesis. However, the downstream targets of VEGF in endothelial cells (ECs) are not entirely clarified. Survey of downstream targets of VEGF in human ECs identified a number of genes, including Down syndrome candidate region 1 (DSCR1). Here, we confirmed the inducible expression of DSCR1 in ECs by Northern and Western blottings. Moreover, VEGF-stimulated induction of DSCR1 was blocked by anti-VEGF receptor-2 monoclonal antibody (mAb), or the specific calcineurin inhibitors cyclosporin A and FK506. The expression of DSCR1 in ECs of neovessels was further shown by immunohistochemical analysis. We therefore examined whether DSCR1 played any roles in angiogenesis. The specific downregulation of DSCR1 expression by antisense oligonucleotide (AS-ODN) inhibited VEGF-stimulated migration of ECs as well as angiogenesis in vivo. AS-ODN inhibited the spreading of ECs on vitronectin, as well as on the immobilized anti-αvβ3 mAb, but not on anti-αvβ5 mAb. Moreover, AS-ODN inhibited tyrosine phosphorylation of focal adhesion kinase when ECs were plated on a vitronectin-coated dish. Immunoprecipitation followed by Western blotting showed the coimmunoprecipitation of DSCR1 and integrin αvβ3. These results suggest that DSCR1 is involved in angiogenesis by regulating adhesion and migration of ECs via the interaction with integrin αvβ3.

Down Syndrome Candidate Region 1-like 1 (DSCR1-L1) Mimics the Inhibitory Effects of DSCR1 on Calcineurin Signaling in Endothelial Cells and Inhibits Angiogenesis

“…Prior studies have demonstrated that DSCR1 inhibits endothelial cell proliferation, migration, and tube formation in vitro (14)(15)(16)(17). To confirm these findings and to compare the effects of DSCR1 to DSCR1-L1, HUVEC were infected with our retroviral construct containing the DSCR1 gene (pMX.myc.DSCR1).…”

“…Activated calcineurin dephosphorylates and induces the nuclear import of NF-ATc1 and NF-ATc2 in endothelial cells, resulting in the expression of angiogenesis-related genes including Cox-2 and E-selectin (11)(12)(13). Studies have demonstrated that DSCR1 is upregulated in response to VEGF in a calcineurin-dependent manner (14)(15)(16)(17), and that over-expression of DSCR1 in endothelial cells inhibits NF-AT translocation, NF-AT-mediated gene expression, and endothelial cell function (14)(15)(16)(17).…”

Luteinization of porcine preovulatory follicles leads to systematic changes in follicular gene expression

“…Two of the enzymes in this group (MAOB and LAP3) are vasodilatory; monoamine oxidase metabolizes vasoactive amines to inactive forms and is found in high amounts within the preovulatory follicle and corpus luteum (Yoshimoto et al 1986) and LAP3 (also known as cysteine amino peptidase or oxitocinase) metabolizes oxytocin, vasopressin, and angiotensin III (Tsujimoto & Hattori 2005) and is a marker for malignant gynecological tumors (Ino et al 2004). The importance of VEGF in the angiogenic cascade that occurs in preovulatory luteinized follicles is well documented (Tamanini & De Ambrogi 2004) and DSCR1, a differentially expressed gene shown here, is an early response gene for VEGF (Iizuka et al 2004). PLANH1, the primary inhibitor of plasminogen activator that has been implicated in tumor growth and cellular remodeling (Stefansson et al 2003) and annexin a2 (ANXA2), a plasminogen-plasminogen activator co-receptor with angiogenic properties (Kim & Hajjar 2002), were also found to be differentially expressed.…”

Two key genes closely implicated with the neuropathological characteristics in Down syndrome: DYRK1A and RCAN1

“…Although RCAN1-4 is induced by an angiogenic factor VEGF, the overexpression of RCAN1-4 attenuated tube formation and cell cycle progression of ECs, thus inhibiting angiogenesis (70,93,94). When RCAN1 was knocked down, VEGF-stimulated migration of ECs and angiogenesis was also inhibited (95). In the previous model of cardiac hypertrophy and angiogenesis, RCAN1 deficiency inhibited calcineurin.…”