S everal prevalent diseases are associated with abnormal angiogenesis and formation of a pathological neovasculature (PNV), notably cancers with solid tumors, diabetic retinopathy, and the exudative (wet) form of age-related macular degeneration (AMD). Two procedures have been described as potential treatments for PNV-associated diseases, an antiangiogenesis protocol to inhibit angiogenesis (1, 2) and an anti-PNV protocol to destroy selectively the PNV (3-6). Because a PNV usually has formed by the time the disease is diagnosed, destruction of the PNV probably will be necessary to achieve optimal therapeutic efficacy. Here we report the results of testing an anti-PNV procedure in a mouse model that simulates exudative AMD. The procedure involves administering a chimeric antibody-like molecule, called an Icon, which binds with high affinity and specificity to the receptor tissue factor (TF). TF is expressed on endothelial cells lining the luminal surface of a PNV but not of a normal vasculature (7, 8), thus providing a specific and accessible therapeutic target. The Icon is composed of factor VII (fVII), the natural ligand for TF, conjugated to the Fc domain of an IgG1 Ig. The Icon functions as an anti-TF antibody, with higher affinity and specificity than can be achieved with an anti-TF antibody. The TF-Icon complex activates a potent cytolytic immune attack mediated by natural killer cells and complement (9). Cytolysis of endothelial cells of the PNV, and possibly of other cells in the wall of a leaky PNV vessel that express TF, results in selective destruction of the PNV, as demonstrated in mouse models of solid tumors (3, 4).Exudative AMD involves pathological angiogenesis that originates from the choroid beneath the retina to form a PNV, called a choroidal neovasculature (CNV), containing abnormal blood vessels that can leak fluid and bleed, hence the name exudative AMD. The fluid and blood released from the CNV can damage the structure and function of the overlying retina, usually in the central macular area, leading to the loss of central vision. Several procedures are currently used to treat submacular CNV, notably photodynamic therapy (10), submacular surgery (11, 12), and macular translocation (13-15). However, most treated patients do not show visual improvement, and the surgical procedures can cause serious complications. The severe visual disability caused by AMD and the lack of an adequate treatment for the disease has motivated a search for new therapeutic strategies. Here we show that formation of a laser-induced CNV can be prevented at an early stage in a mouse model, by i.v. or intraocular (i.o.) injections of either an adenoviral vector encoding the Icon or Icon protein.
Materials and MethodsAnimals. C57BL͞6 mice 4-6 weeks old were purchased from The Jackson Laboratory, and pigs 10-12 weeks old were purchased from Professional Veterinary Research (Brownstown, IN). The animals were maintained in accord with the guidelines established by the Committee on Animals at the University of Louisville Me...