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Objective- Pathological neovascularization is crucial for progression and morbidity of serious diseases such as cancer, diabetic retinopathy, and age-related macular degeneration. While mechanisms of ongoing pathological neovascularization have been extensively studied, the initiating pathological vascular remodeling (PVR) events, which precede neovascularization remains poorly understood. Here, we identify novel molecular and cellular mechanisms of preneovascular PVR, by using the adult choriocapillaris as a model. Approach and Results- Using hypoxia or forced overexpression of VEGF (vascular endothelial growth factor) in the subretinal space to induce PVR in zebrafish and rats respectively, and by analyzing choriocapillaris membranes adjacent to choroidal neovascular lesions from age-related macular degeneration patients, we show that the choriocapillaris undergo robust induction of vascular intussusception and permeability at preneovascular stages of PVR. This PVR response included endothelial cell proliferation, formation of endothelial luminal processes, extensive vesiculation and thickening of the endothelium, degradation of collagen fibers, and splitting of existing extravascular columns. RNA-sequencing established a role for endothelial tight junction disruption, cytoskeletal remodeling, vesicle- and cilium biogenesis in this process. Mechanistically, using genetic gain- and loss-of-function zebrafish models and analysis of primary human choriocapillaris endothelial cells, we determined that HIF (hypoxia-induced factor)-1α-VEGF-A-VEGFR2 signaling was important for hypoxia-induced PVR. Conclusions- Our findings reveal that PVR involving intussusception and splitting of extravascular columns, endothelial proliferation, vesiculation, fenestration, and thickening is induced before neovascularization, suggesting that identifying and targeting these processes may prevent development of advanced neovascular disease in the future. Visual Overview- An online visual overview is available for this article.

Purpose: Scavenging of VEGF by specific antibodies is a promising way to treat ocular conditions connected with neovascularization. Intravitreal injections of Avastin (bevacizumab) are performed frequently as a treatment of such conditions. In this study, the authors examine whether the retinal function in wild-type mice is affected by an intravitreal injection of Avastin., Methods: Electroretinography was performed in four different experimental groups of wild-type C57BL/6 mice before treatment and 1, 4, 12, and 25 days afterwards. The first group was injected intravitreally with BSS, the second one received injections of a vehicle solution, and the third group was injected with the commercial Avastin solution. In a fourth group, sham surgery was performed. Immunohistochemistry was performed in some eyes to evaluate penetration of the bevacizumab molecule through the retina., Results: In all four groups, a similar behavior of the ERG parameters could be detected. One day after the injections, the amplitudes showed a clear decrease. Later on, they recovered gradually. No difference could be seen between eyes injected with Avastin or vehicle solution. Bevacizumab immunoreactivity was already present in the whole retina half an hour after the intravitreal injection and was not detectable 25 days later. Moreover, binding of bevacizumab to endogenous mouse VEGF could be shown., Conclusions: Based on the electroretinographic findings, the authors conclude that bevacizumab does not have any toxic effects on the mouse retina and its function. The bevacizumab molecule penetrates the retina quickly. Therefore, it can act safely and very quickly, also in deeper retinal layers after its injection.

Background: Transplantation of autologous iris pigment epithelium (IPE) into the subretinal space has been suggested as one approach for the treatment of age-related macular degeneration. Autologous rabbit IPE cells were transplanted to the subretinal space to define the technique of transplantation and examine the survival of the transplanted cells., Methods: Autologous IPE cells were harvested by iridectomy and transplanted directly to the subretinal space of the fellow eye in 25 rabbits, using the parsplana approach. Animals were killed over a period of 5 months, and the retinas were examined morphologically by light and electron microscopy., Results: Autologous IPE cells survived and formed a polarized monolayer above the retinal pigment epithelium in the subretinal space, with apical microvilli adjacent to photoreceptors. Fragments of phagocytosed photoreceptor rod outer segments were observed in phagosomes in the cytoplasm of IPE cells. Adjacent rod outer segments remained healthy throughout the experimental period. No signs of a cell-mediated immunologic response were observed., Conclusions: Our results show that in rabbits, autologous IPE cells transplanted to the subretinal space survive and do not adversely affect the photoreceptors. These results suggest that in humans, IPE cells might provide a substitute for retinal pigment epithelium cells as autologous transplants for the treatment of age-related macular degeneration.

Pancreata of normoglycemic diabetes-prone (dp) and diabetes-resistant (dr) BB rats and normal Wistar rats were screened for macrophage infiltration by immunohistochemistry and by electron microscopy. Inflammatory macrophages were found in the endocrine as well as the exocrine part of the pancreata from dp BB rats. In the exocrine tissue they had a different phenotype (ED1+, ED2+, W3/25+, Ox17+) from that found in pancreata from dr BB and Wistar rats (predominantly ED1-, ED2+, W3/25+, Ox17-). The number of macrophages in exocrine portions of pancreata from the various rat strains were not different. By electron microscopy scattered macrophage-associated tissue lesions and phagocytosis of cell debris were found throughout the exocrine tissue and in islets of dp BB rats. Such lesions were low or absent in biopsies of animals that later did not develop diabetes. We conclude that macrophage-mediated cytotoxicity during the early phases of diabetes development in BB rats is not restricted to islets, but is a generalized, pan-pancreatic event.