Correlation of Histologic 2-Dimensional Reconstruction and Confocal Scanning Laser Microscopic Imaging of Choroidal Neovascularization in Eyes With Age-Related Maculopathy

Abstract

To topographically localize vascular channels, macrophages, and retinal pigment epithelium and other components of choroidal neovascularization (CNV) associated with age-related maculopathy. Methods: Two postmortem eyes with age-related maculopathy and CNV were evaluated. The formalin-fixed CNV complex was excised and processed for confocal scanning laser microscopy including immunostaining for factor VIII-related antigen and incubation with Ig fluorescein isothiocyanate. After confocal microscopy, the specimens were serial step sectioned, stained, and 2-dimensional topographic reconstructions were made. The confocal images were compared with the 2-dimensional reconstructions. Results: Both specimens contained central disciform scars surrounded by areas of intact retinal pigment epithelium. The first specimen was more atrophic and contained fewer choroidal neovascular channels than the second specimen. The topographic arrangement of the CNV and retinal pigmentepithelialchangesintheconfocalimagescorresponded with the 2-dimensional reconstructions. Macrophages were concentrated around areas of vascularization. Conclusion: Confocal scanning laser microscopy of excised CNV simulates fluorescein angiography and topographic localization of the components of CNV provides insight into the pathogenesis of CNV.

Targeting cell-type-specific, choroid-peripheral immune signaling to treat age-related macular degeneration

“…Previous studies have shown that monocyte-derived macrophages are an abundant immune cell localized to CNV lesions. 17 , 22 , 32 , 33 Consistent with this, we found that there were many CCR2+ monocyte-derived macrophages in close proximity to CYR61 expressed by endothelial cells in choroidal neovascular lesions ( Figure 4 B). Therefore, we hypothesized that the CYR61 produced by endothelial cells in CNV lesions could signal to and recruit or activate monocyte-derived macrophages.…”

“…More specifically, we found that CCR2+ monocyte-derived macrophages were present in the CYR61-rich neovascular lesion ( Figure 4 B), consistent with previous studies. 17 , 22 , 32 , 33 Taken together, these findings support that CYR61 expressed specifically by endothelial cells in CNV lesions could signal to nearby immune cells, including monocyte-derived macrophages.…”

Targeting cell-type-specific, choroid-peripheral immune signaling to treat age-related macular degeneration

“…Previous studies have shown that monocyte-derived macrophages are an abundant immune cell localized to CNV lesions. 17 , 22 , 32 , 33 Consistent with this, we found that there were many CCR2+ monocyte-derived macrophages in close proximity to CYR61 expressed by endothelial cells in choroidal neovascular lesions ( Figure 4 B). Therefore, we hypothesized that the CYR61 produced by endothelial cells in CNV lesions could signal to and recruit or activate monocyte-derived macrophages.…”

“…More specifically, we found that CCR2+ monocyte-derived macrophages were present in the CYR61-rich neovascular lesion ( Figure 4 B), consistent with previous studies. 17 , 22 , 32 , 33 Taken together, these findings support that CYR61 expressed specifically by endothelial cells in CNV lesions could signal to nearby immune cells, including monocyte-derived macrophages.…”

Anti‐VEGF Aptamer (Pegaptanib) Therapy for Ocular Vascular Diseases

“…As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization. 58,79,80 In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups. [88][89][90][91] While the specific role of VEGF in attracting the monocyte/macrophage cells has not been established in these experimental models, it is noteworthy that VEGF is a chemoattractant for these cells, 92 with VEGF 165 again proving to be especially potent in this regard.…”

“…Finally, one intriguing feature of pathological neovascularization 74 is the potential role of macrophages in enhancing this process. As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization 58,79,80 . In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups 88–91 .…”

Visualizing Retinal Pigment Epithelium Phenotypes in the Transition to Geographic Atrophy in Age-Related Macular Degeneration

“…While not contradicting well-established roles of macrophages 67, 68 and inner-retina-derived microglia 61, 69, 70 in advanced AMD, our data point to a role for transdifferentiated RPE that remains to be determined. Further, whether treatments should promote survival or suppression of these cells is unknown.…”

Targeting cell-type-specific, choroid-peripheral immune signaling to treat age-related macular degeneration

“…Previous studies have shown that monocyte-derived macrophages are an abundant immune cell localized to CNV lesions. 17 , 22 , 32 , 33 Consistent with this, we found that there were many CCR2+ monocyte-derived macrophages in close proximity to CYR61 expressed by endothelial cells in choroidal neovascular lesions ( Figure 4 B). Therefore, we hypothesized that the CYR61 produced by endothelial cells in CNV lesions could signal to and recruit or activate monocyte-derived macrophages.…”

“…More specifically, we found that CCR2+ monocyte-derived macrophages were present in the CYR61-rich neovascular lesion ( Figure 4 B), consistent with previous studies. 17 , 22 , 32 , 33 Taken together, these findings support that CYR61 expressed specifically by endothelial cells in CNV lesions could signal to nearby immune cells, including monocyte-derived macrophages.…”

Anti‐VEGF Aptamer (Pegaptanib) Therapy for Ocular Vascular Diseases

“…As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization. 58,79,80 In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups. [88][89][90][91] While the specific role of VEGF in attracting the monocyte/macrophage cells has not been established in these experimental models, it is noteworthy that VEGF is a chemoattractant for these cells, 92 with VEGF 165 again proving to be especially potent in this regard.…”

“…Finally, one intriguing feature of pathological neovascularization 74 is the potential role of macrophages in enhancing this process. As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization 58,79,80 . In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups 88–91 .…”

Visualizing Retinal Pigment Epithelium Phenotypes in the Transition to Geographic Atrophy in Age-Related Macular Degeneration

“…While not contradicting well-established roles of macrophages 67, 68 and inner-retina-derived microglia 61, 69, 70 in advanced AMD, our data point to a role for transdifferentiated RPE that remains to be determined. Further, whether treatments should promote survival or suppression of these cells is unknown.…”

Targeting cell-type-specific, choroid-peripheral immune signaling to treat age-related macular degeneration

“…Previous studies have shown that monocyte-derived macrophages are an abundant immune cell localized to CNV lesions. 17 , 22 , 32 , 33 Consistent with this, we found that there were many CCR2+ monocyte-derived macrophages in close proximity to CYR61 expressed by endothelial cells in choroidal neovascular lesions ( Figure 4 B). Therefore, we hypothesized that the CYR61 produced by endothelial cells in CNV lesions could signal to and recruit or activate monocyte-derived macrophages.…”

“…More specifically, we found that CCR2+ monocyte-derived macrophages were present in the CYR61-rich neovascular lesion ( Figure 4 B), consistent with previous studies. 17 , 22 , 32 , 33 Taken together, these findings support that CYR61 expressed specifically by endothelial cells in CNV lesions could signal to nearby immune cells, including monocyte-derived macrophages.…”

Anti‐VEGF Aptamer (Pegaptanib) Therapy for Ocular Vascular Diseases

“…As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization. 58,79,80 In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups. [88][89][90][91] While the specific role of VEGF in attracting the monocyte/macrophage cells has not been established in these experimental models, it is noteworthy that VEGF is a chemoattractant for these cells, 92 with VEGF 165 again proving to be especially potent in this regard.…”

“…Finally, one intriguing feature of pathological neovascularization 74 is the potential role of macrophages in enhancing this process. As described above, macrophages are commonly seen in clinical cases of pathological ocular vascularization 58,79,80 . In the retinal ischemia model system, 74 inactivation of monocyte lineage cells dramatically inhibited the formation of the pathological neovascularization, and similar findings have been reported by several other groups 88–91 .…”

Visualizing Retinal Pigment Epithelium Phenotypes in the Transition to Geographic Atrophy in Age-Related Macular Degeneration

“…While not contradicting well-established roles of macrophages 67, 68 and inner-retina-derived microglia 61, 69, 70 in advanced AMD, our data point to a role for transdifferentiated RPE that remains to be determined. Further, whether treatments should promote survival or suppression of these cells is unknown.…”