Serum-deprivation response of ARPE-19 cells; expression patterns relevant to age-related macular degeneration

PLoS One. 2024 Sep 26;19(9):e0293383. doi: 10.1371/journal.pone.0293383. eCollection 2024.

Abstract

ARPE-19 cells are derived from adult human retinal pigment epithelium (RPE). The response of these cells to the stress of serum deprivation mimics some important processes relevant to age-related macular degeneration (AMD). Here we extend the characterization of this response using RNASeq and EGSEA gene set analysis of ARPE-19 cells over nine days of serum deprivation. This experiment confirmed the up-regulation of cholesterol and lipid-associated pathways that increase cholesterol levels in these cells. The gene expression analysis also identified other pathways relevant to AMD progression. There were significant changes in extracellular matrix gene expression, notably a switch from expression of collagen IV, a key component of Bruch's membrane (part of the blood-retina barrier), to expression of a fibrosis-like collagen type I matrix. Changes in the expression profile of the extracellular matrix led to the discovery that amelotin is induced in AMD and is associated with the development of the calcium deposits seen in late-stage geographic atrophy. The transcriptional profiles of other pathways, including inflammation, complement, and coagulation, were also modified, consistent with immune response patterns seen in AMD. As previously noted, the cells resist apoptosis and autophagy but instead initiate a gene expression pattern characteristic of senescence, consistent with the maintenance of barrier function even as other aspects of RPE function are compromised. Other differentially regulated genes were identified that open new avenues for investigation. Our results suggest that ARPE-19 cells maintain significant stress responses characteristic of native RPE that are informative for AMD. As such, they provide a convenient system for discovery and for testing potential therapeutic interventions.

MeSH terms

  • Cell Line
  • Cholesterol / metabolism
  • Culture Media, Serum-Free / pharmacology
  • Extracellular Matrix / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Humans
  • Macular Degeneration* / genetics
  • Macular Degeneration* / metabolism
  • Macular Degeneration* / pathology
  • Retinal Pigment Epithelium* / metabolism
  • Retinal Pigment Epithelium* / pathology
  • Transcriptome

Substances

  • Culture Media, Serum-Free
  • Cholesterol

Grants and funding

This research was supported by the Intramural Research Program of the NIH, National Eye Institute.