Description

Age-related macular degeneration (AMD) is a leading cause of blindness worldwide. Unfortunately, the early stages of this disease are poorly understood, which has led to limited treatment options. Investigating normal changes in tissues eventually affected by AMD can further elucidate the mechanisms of disease progression and lead to novel therapeutic targets. The primary cell layer affected in AMD is the retinal pigment epithelium (RPE), which forms the outer blood-retinal barrier (oBRB). Beneath the RPE lies Bruch’s membrane, a proteinaceous layer that naturally thickens and stiffens with age. These changes to Bruch’s membrane are also implicated in RPE dysfunction and AMD progression. To investigate the relationship between normal, age-related changes in Bruch’s membrane and AMD development, we engineered a tunable in vitro model of Bruch’s membrane to support primary porcine RPE cells. We performed transepithelial electrical resistance (TEER) measurements, viability assays, morphological analysis, immunocytochemistry, and enzyme-linked immunosorbent assays (ELISA) to evaluate monolayer integrity and angiogenic factor expression. Cells cultured on our aged model exhibited changes similar to those seen in AMD, including reduced monolayer integrity, the formation of sub-RPE deposits, and eventual cell death. Notably, apolipoprotein E (ApoE), a known drusen component and Alzheimer’s disease marker, was overexpressed prior to deposit accumulation and cell death. Regions of ApoE overexpression corresponded with disrupted expression of zonula occludens-1, a junctional protein. While most angiogenic factors remained unchanged, tissue inhibitor of metalloproteinases-1 (TIMP-1) was transiently overexpressed before cell death. These findings suggest that ApoE and TIMP-1 may play key roles in early AMD pathogenesis and represent potential targets for future therapeutic intervention

Author ORCID Identifier

Elizabeth Vargis  https://orcid.org/0000-0003-3141-9317

Document Type

Dataset

DCMI Type

Dataset

File Format

.xlsx

Publication Date

1-20-2026

Funder

National Institutes of Health;

Utah State University Alzheimer's Disease and Dementia Research Center;

Utah State University's Office of Research;

Utah NASA Space Grant Consortium;

BrightFocus Foundation;

Retinal Research Foundation

Publisher

Utah State University

Award Number

National Institutes of Health EY028732; BrightFocus Foundation M2019109

Methodology

Raw data is provided in this file. TEER data is organized into two sections, all of which include raw resistance values in Ohms. Data are separated into a preliminary section, which includes regular measurements that were converted into the values seen in the published manuscript, and the later section, which includes initial mature readings and a final reading taken at the end of the culture period. ELISA data includes concentrations for relevant angiogenic factors on various membranes at various time points. Viability data includes absorbance readings that were used to estimate percent viability in the manuscript.

Referenced by

D. Weatherston, J. A. Jones, E. Vargis, Changes in ApoE and TIMP-1 Expression Correlate with Outer Blood-Retinal Barrier Disruption in an In Vitro Model of Retinal Aging, Geroscience

Language

eng

Code Lists

see README

Disciplines

Biological Engineering

License

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.

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a4de9e8347316b8b2ab5d9df55f4b385

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Additional Files

Weaterhston_DigitalCommons_README.txt (6 kB)
md5 checksum: d730db3cb4a19a5459146ca2a67ee2da

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