Role of Endomucin in Hypoxia-Induced Retinopathy of Prematurity

Retinopathy of prematurity (ROP) is a major cause of blindness among premature, low birth weight infants as a result of pathological angiogenesis. Angiogenesis, the growth of new blood vessels from preexisting vessels, occurs in the veins and capillaries of the body. The process is highly regulated during early development and maturation. However, under abnormal conditions such as a decrease in oxygen levels or hypoxia, angiogenesis can become dysregulated and pathogenic. Currently, the best treatment for ROP is laser therapy, which does not significantly improve vision. Alternatively, glycoproteins are believed to play an important role in angiogenesis. Endomucin (EMCN), a glycoprotein, has been shown to be expressed by the venous and capillary endothelium. EMCN is believed to be associated with angiogenesis and could be a potential target for treatment of ROP. Thus, we hypothesize that EMCN is regulated by hypoxia and plays an important role in pathological angiogenesis. Human retinal endothelial cells (HRECs), representative of endothelial cells involved in retinal angiogenesis, were deprived of oxygen using a hypoxia chamber. We established the optimal oxygen dosage, determined the optimal cell density, and monitored EMCN expression at different time points after exposure to hypoxia. Changes in gene expression in response to hypoxia were compared to control cells. Our preliminary data indicates that EMCN is regulated by hypoxia. Currently, we are investigating whether EMCN has similar effects in regulating revascularization in vivo. Taken together, our study indicates a novel role for EMCN during hypoxia-induced angiogenesis which may serve as a therapeutic target

VEGF\u3csub\u3e164\u3c/sub\u3e-Mediated Inflammation is Required for Pathological, but Not Physiological, Ischemia-Induced Retinal Neovascularization

Hypoxia-induced VEGF governs both physiological retinal vascular development and pathological retinal neovascularization. In the current paper, the mechanisms of physiological and pathological neovascularization are compared and contrasted. During pathological neovascularization, both the absolute and relative expression levels for VEGF164 increased to a greater degree than during physiological neovascularization. Furthermore, extensive leukocyte adhesion was observed at the leading edge of pathological, but not physiological, neovascularization. When a VEGF164-specific neutralizing aptamer was administered, it potently suppressed the leukocyte adhesion and pathological neovascularization, whereas it had little or no effect on physiological neovascularization. In parallel experiments, genetically altered VEGF164-deficient (VEGF120/188) mice exhibited no difference in physiological neovascularization when compared with wild-type (VEGF+/+) controls. In contrast, administration of a VEGFR-1/Fc fusion protein, which blocks all VEGF isoforms, led to significant suppression of both pathological and physiological neovascularization. In addition, the targeted inactivation of monocyte lineage cells with clodronate-liposomes led to the suppression of pathological neovascularization. Conversely, the blockade of T lymphocyte–mediated immune responses with an anti-CD2 antibody exacerbated pathological neovascularization. These data highlight important molecular and cellular differences between physiological and pathological retinal neovascularization. During pathological neovascularization, VEGF164 selectively induces inflammation and cellular immunity. These processes provide positive and negative angiogenic regulation, respectively. Together, new therapeutic approaches for selectively targeting pathological, but not physiological, retinal neovascularization are outlined

VEGF and TGF-beta are required for the maintenance of the choroid plexus and ependyma.

Although the role of vascular endothelial growth factor (VEGF) in developmental and pathological angiogenesis is well established, its function in the adult is less clear. Similarly, although transforming growth factor (TGF) beta is involved in angiogenesis, presumably by mediating capillary (endothelial cell [EC]) stability, its involvement in quiescent vasculature is virtually uninvestigated. Given the neurological findings in patients treated with VEGF-neutralizing therapy (bevacizumab) and in patients with severe preeclampsia, which is mediated by soluble VEGF receptor 1/soluble Fms-like tyrosine kinase receptor 1 and soluble endoglin, a TGF-beta signaling inhibitor, we investigated the roles of VEGF and TGF-beta in choroid plexus (CP) integrity and function in adult mice. Receptors for VEGF and TGF-beta were detected in adult CP, as well as on ependymal cells. Inhibition of VEGF led to decreased CP vascular perfusion, which was associated with fibrin deposition. Simultaneous blockade of VEGF and TGF-beta resulted in the loss of fenestrae on CP vasculature and thickening of the otherwise attenuated capillary endothelium, as well as the disappearance of ependymal cell microvilli and the development of periventricular edema. These results provide compelling evidence that both VEGF and TGF-beta are involved in the regulation of EC stability, ependymal cell function, and periventricular permeability