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Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria

J Y Ahn1, S Datta2, E Bandeira1

  • 1Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America.

Biochimica Et Biophysica Acta. General Subjects
|April 3, 2020
PubMed
Abstract

Insights

Mitochondrial dysfunction in retinal pigment epithelial cells can be detected by elevated levels of miR-494-3p in extracellular vesicles. This finding suggests EV miR-494-3p as a potential biomarker for RPE health and retinal degeneration.

Area of Science:

  • Cell Biology
  • Mitochondrial Biology
  • Extracellular Vesicles

Background:

  • Mitochondrial function in retinal pigment epithelial (RPE) cells is linked to extracellular vesicle (EV) formation and release.
  • Lysosomal and exocytotic pathways manage intracellular material, including mitochondrial fragments, influencing EV content.
  • Impaired RPE mitochondria are hypothesized to release EVs containing specific mitochondrial microRNAs (miRNAs).

Purpose of the Study:

  • To identify specific miRNAs within mitochondria and released EVs in RPE cells.
  • To investigate the role of these miRNAs in mitochondrial function and EV release.
  • To determine if mitochondrial dysfunction is reflected in the miRNA cargo of EVs.

Main Methods:

  • ARPE-19 cells were screened for mitochondrial miRNAs present in EVs.
  • Cells were treated with rotenone to induce mitochondrial injury.
  • EVs were characterized by size, biomarkers (CD81, CD63, syntenin-1), miRNA cargo, and concentration.
  • miRNA localization to mitochondria and its functional impact were assessed via knockdown experiments.

Main Results:

  • miR-494-3p was found to be enriched in ARPE-19 cell mitochondria.
  • Knockdown of miR-494-3p impaired mitochondrial function, reducing ATP production and membrane potential.
  • Rotenone treatment increased EV release and elevated miR-494-3p levels in released EVs.

Conclusions:

  • miR-494-3p regulates mitochondrial function in ARPE-19 cells.
  • Elevated miR-494-3p in EVs signifies compromised mitochondrial capacity within RPE cells.
  • EV miR-494-3p shows potential as a biomarker for RPE mitochondrial dysfunction and retinal degeneration.
Keywords:
Age-related macular degenerationExtracellular vesiclesMitochondriaRetinal pigment epithelial cellsmiR-494-3pmiRNA

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