Choriocapillaris Degeneration in Geographic Atrophy

Sohn, Elliott H.; Flamme-Wiese, Miles J.; Whitmore, S. Scott; Workalemahu, Grefachew; Marneros, Alexander G.; Boese, Erin A.; Kwon, Young H.; Wang, Kai; Abràmoff, Michael D.; Tucker, Budd A.; Stone, Edwin M.; Mullins, Robert F.

doi:10.1016/j.ajpath.2019.04.005

Cited by 51 publications

(36 citation statements)

References 52 publications

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“…There is mounting evidence that damage to the choroidal vasculature, the blood supply for the retinal pigment epithelium and outer retina, plays a key role in the pathogenesis of age-related macular degeneration (AMD) [1][2][3][4][5] and is altered in other diseases such as diabetic retinopathy, central serous retinopathy, and myopia. [6][7][8][9][10] There are relatively robust methods for evaluating morphometric abnormalities of the choroid in humans using histology and fluorescent labeling of donor eyes that can discriminate between intact and ghost vessels 1,2,[11][12][13][14] as well as in vivo imaging techniques such as optical coherence tomography and optical coherence tomography angiography. [15][16][17] Such rapid and reliable methods for evaluating the choroid in mice are more challenging especially for whole mount, en face visualization.…”

Section: Introductionmentioning

confidence: 99%

Visualization of Mouse Choroidal and Retinal Vasculature Using Fluorescent Tomato Lectin Perfusion

Jiao

Adler

Liu

et al. 2020

Trans. Vis. Sci. Tech.

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Purpose: To develop a reliable and simplified method to assess choroid and retinal vasculature on whole mount and cross sections in mice using tomato lectin (TL; Lycopersicon esculentum). Methods: Albino mice (n = 27) received 1 mg/mL of TL (conjugated to Dylight-594) intravascularly through the tail vein, jugular vein, or cardiac left ventricle. Whole mounts of the retina and choroid were evaluated using fluorescence microscopy. Perfusion with GSL-IB4 conjugated to Dylight-594 and fluorescein isothiocyanate was performed to compare against labeling with TL. Co-labeling of choroidal endothelial cells with perfused TL on cross-sections with antibodies directed against the choriocapillarisrestricted endothelial cell marker CA4 was performed. The percentage of perfused choroidal and retinal vessels was assessed semiquantitatively. One mouse was subjected to thermal laser damage before perfusion to cause retinal and choroidal vasculature ablation. Results: Intravascular injection of TL led to consistent, robust labeling of retinal and choroidal vascular walls. On cross-sections, choriocapillaris was co-labeled with CA4 and TL. On flat mount, TL perfusion resulted in better labeling of choroidal vessels using tail/jugular vein injection compared with cardiac perfusion (P < .01). More consistent labeling of the choroidal and retinal vascular trees was observed with TL than with GSL-IB4. Vascular damage caused by laser ablation was detected readily using this method. Conclusions: TL injection intravascularly can reliably label normal and ablated choroid and retinal vasculature in mouse in a quick, simple manner. Translational Relevance: These data will help to facilitate modeling in rodents for diseases such as age-related macular degeneration, diabetes, and other ischemic/angiogenic processes that can also be used for treatment evaluation.

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Section: Introductionmentioning

confidence: 99%

Visualization of Mouse Choroidal and Retinal Vasculature Using Fluorescent Tomato Lectin Perfusion

Jiao

Adler

Liu

et al. 2020

Trans. Vis. Sci. Tech.

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“…Therefore the stated values might not be absolute, which has to be accepted as a limitation. The involvement of choroidal and choriocapillary vessel changes in the pathophysiology of GA was reported 4,43 ; however, this additional issue was not part of this study's investigation. Future studies will have to combine vascular changes with OCT structural findings.…”

Section: Discussionmentioning

confidence: 84%

Subretinal Drusenoid Deposits and Photoreceptor Loss Detecting Global and Local Progression of Geographic Atrophy by SD-OCT Imaging

Reiter

Told

Schranz

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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To investigate the impact of subretinal drusenoid deposits (SDD) and photoreceptor integrity on global and local geographic atrophy (GA) progression. METHODS. Eighty-three eyes of 49 patients, aged 50 years and older with GA secondary to age-related macular degeneration (AMD), were prospectively included in this study. Participants underwent spectral-domain optical coherence tomography (SD-OCT) and fundus autofluorescence (FAF) imaging at baseline and after 12 months. The junctional zone and presence of SDD were delineated on SD-OCT and FAF images. Linear mixed models were calculated to investigate the association between GA progression and the junctional zone area, baseline GA area, age, global and local presence of SDD and unifocal versus multifocal lesions. RESULTS. The area of the junctional zone was significantly associated with the progression of GA, both globally and locally (all P < 0.001). SDD were associated with faster growth in the overall model (P = 0.039), as well as in the superior-temporal (P = 0.005) and temporal (P = 0.002) sections. Faster progression was associated with GA baseline area (P < 0.001). No difference was found between unifocal and multifocal lesions (P > 0.05). Age did not have an effect on GA progression (P > 0.05). CONCLUSIONS. Photoreceptor integrity and SDD are useful for predicting global and local growth in GA. Investigation of the junctional zone is merited because this area is destined to become atrophic. Photoreceptor loss visible on SD-OCT might lead to new structural outcome measurements visible before irreversible loss of retinal pigment epithelium occurs.

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“…As indicated above, choriocapillaris loss occurs early in the pathogenesis of AMD, often preceding degeneration of the overlying RPE and photoreceptor cells [ 8 – 10 ]. As such, autologous iPSC-derived CEC replacement early in the disease course could be advantageous in slowing AMD progression, including the atrophic dry form of the disease, which currently has no treatment.…”

Section: Discussionmentioning

confidence: 99%

“…Immunohistochemical and gene expression analyses of human donor eyes demonstrate that endothelial cells lining the choriocapillaris are lost prior to RPE degeneration, creating empty lumens of extracellular matrix termed ghost vessels [ 8 , 9 ]. Morphometric analysis of the choroidal vasculature in eyes with AMD further supports choriocapillaris vessel loss in early AMD, but similar differences are not readily apparent in the deeper choroidal vessels, suggesting vascular dropout may occur specifically in the capillaries [ 10 ]. Standard methods of visualizing choroidal vascular anatomy in vivo include indocyanine green angiography, but this test is invasive, requiring an intravenous injection of dye [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Stepwise differentiation and functional characterization of human induced pluripotent stem cell-derived choroidal endothelial cells

et al. 2020

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Background Endothelial cells (ECs) are essential regulators of the vasculature, lining arteries, veins, and capillary beds. While all ECs share a number of structural and molecular features, heterogeneity exists depending on their resident tissue. ECs lining the choriocapillaris in the human eye are lost early in the pathogenesis of age-related macular degeneration (AMD), a common and devastating form of vision loss. In order to study the mechanisms leading to choroidal endothelial cell (CEC) loss and to develop reagents for repairing the choroid, a reproducible in vitro model, which closely mimic CECs, is needed. While a number of protocols have been published to direct induced pluripotent stem cells (iPSCs) into ECs, the goal of this study was to develop methods to differentiate iPSCs into ECs resembling those found in the human choriocapillaris specifically. Methods We transduced human iPSCs with a CDH5p-GFP-ZEO lentiviral vector and selected for transduced iPSCs using blasticidin. We generated embryoid bodies (EBs) from expanded iPSC colonies and transitioned from mTESR™1 to EC media. One day post-EB formation, we induced mesoderm fate commitment via addition of BMP-4, activin A, and FGF-2. On day 5, EBs were adhered to Matrigel-coated plates in EC media containing vascular endothelial cell growth factor (VEGF) and connective tissue growth factor (CTGF) to promote CEC differentiation. On day 14, we selected for CECs using either zeocin resistance or anti-CD31 MACS beads. We expanded CECs post-selection and performed immunocytochemical analysis of CD31, carbonic anhydrase IV (CA4), and RGCC; tube formation assays; and transmission electron microscopy to access vascular function. Results We report a detailed protocol whereby we direct iPSC differentiation toward mesoderm and utilize CTGF to specify CECs. The CDH5p-GFP-ZEO lentiviral vector facilitated the selection of iPSC-derived ECs that label with antibodies directed against CD31, CA4, and RGCC; form vascular tubes in vitro; and migrate into empty choroidal vessels. CECs selected using either antibiotic selection or CD31 MACS beads showed similar characteristics, thereby making this protocol easily reproducible with or without lentiviral vectors. Conclusion ECs generated following this protocol exhibit functional and biochemical characteristics of CECs. This protocol will be useful for developing in vitro models toward understanding the mechanisms of CEC loss early in AMD.

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Choriocapillaris Degeneration in Geographic Atrophy

Cited by 51 publications

References 52 publications

Visualization of Mouse Choroidal and Retinal Vasculature Using Fluorescent Tomato Lectin Perfusion

Visualization of Mouse Choroidal and Retinal Vasculature Using Fluorescent Tomato Lectin Perfusion

Subretinal Drusenoid Deposits and Photoreceptor Loss Detecting Global and Local Progression of Geographic Atrophy by SD-OCT Imaging

Stepwise differentiation and functional characterization of human induced pluripotent stem cell-derived choroidal endothelial cells

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