Tags

Type your tag names separated by a space and hit enter

Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria.
Biochim Biophys Acta Gen Subj. 2021 04; 1865(4):129598.BB

Abstract

BACKGROUND

Mitochondrial function in retinal pigmented epithelial (RPE) cells and extracellular vesicle (EV) formation/release are related through the lysosomal and exocytotic pathways that process and eliminate intracellular material, including mitochondrial fragments. We propose that RPE cells with impaired mitochondria will release EVs containing mitochondrial miRNAs that reflect the diminished capacity of mitochondria within these cells.

METHODS

We screened ARPE-19 cells for miRNAs that localize to the mitochondria, exhibit biological activity, and are present in EVs released by both untreated cells and cells treated with rotenone to induce mitochondrial injury. EVs were characterized by vesicle size, size distribution, presence of EV biomarkers: CD81, CD63, and syntenin-1, miRNA cargo, and number concentration of EVs released per cell.

RESULTS

We found that miR-494-3p was enriched in ARPE-19 mitochondria. Knockdown of miR-494-3p in ARPE-19 cells decreased ATP production and mitochondrial membrane potential in a dose-dependent manner, and decreased basal oxygen consumption rate and maximal respiratory capacity. Increased number of EVs released per cell and elevated levels of miR-494-3p in EVs released from ARPE-19 cells treated with rotenone were also measured.

CONCLUSIONS

ARPE-19 mitochondrial function is regulated by miR-494-3p. Elevated levels of miR-494-3p in EVs released by ARPE-19 cells indicate diminished capacity of the mitochondria within these cells.

GENERAL SIGNIFICANCE

EV miR-494-3p is a potential biomarker for RPE mitochondrial dysfunction, which plays a central role in non-neovascular age-related macular degeneration, and may be a diagnostic biomarker for monitoring the spread of degeneration to neighboring RPE cells in the retina.

Links

Publisher Full Text (DOI)

Authors+Show Affiliations

Ahn JY
Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America.
Datta S
Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Bandeira E
Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America.
Cano M
Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Mallick E
Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Rai U
Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America.
Powell B
Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Tian J
Biostatistics Center, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America.
Witwer KW
Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Handa JT
Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Paulaitis ME
Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, United States of America. Electronic address: michaelp@jhmi.edu.

MeSH

Cell LineExtracellular VesiclesHumansMacular DegenerationMicroRNAsMitochondriaRetinal Pigment Epithelium

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32240720

Citation

Ahn, J Y., et al. "Release of Extracellular Vesicle miR-494-3p By ARPE-19 Cells With Impaired Mitochondria." Biochimica Et Biophysica Acta. General Subjects, vol. 1865, no. 4, 2021, p. 129598.
Ahn JY, Datta S, Bandeira E, et al. Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria. Biochim Biophys Acta Gen Subj. 2021;1865(4):129598.
Ahn, J. Y., Datta, S., Bandeira, E., Cano, M., Mallick, E., Rai, U., Powell, B., Tian, J., Witwer, K. W., Handa, J. T., & Paulaitis, M. E. (2021). Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria. Biochimica Et Biophysica Acta. General Subjects, 1865(4), 129598. https://doi.org/10.1016/j.bbagen.2020.129598
Ahn JY, et al. Release of Extracellular Vesicle miR-494-3p By ARPE-19 Cells With Impaired Mitochondria. Biochim Biophys Acta Gen Subj. 2021;1865(4):129598. PubMed PMID: 32240720.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Release of extracellular vesicle miR-494-3p by ARPE-19 cells with impaired mitochondria. AU - Ahn,J Y, AU - Datta,S, AU - Bandeira,E, AU - Cano,M, AU - Mallick,E, AU - Rai,U, AU - Powell,B, AU - Tian,J, AU - Witwer,K W, AU - Handa,J T, AU - Paulaitis,M E, Y1 - 2020/03/30/ PY - 2019/10/05/received PY - 2020/01/31/revised PY - 2020/03/09/accepted PY - 2020/4/3/pubmed PY - 2021/6/29/medline PY - 2020/4/3/entrez KW - Age-related macular degeneration KW - Extracellular vesicles KW - Mitochondria KW - Retinal pigment epithelial cells KW - miR-494-3p KW - miRNA SP - 129598 EP - 129598 JF - Biochimica et biophysica acta. General subjects JO - Biochim Biophys Acta Gen Subj VL - 1865 IS - 4 N2 - BACKGROUND: Mitochondrial function in retinal pigmented epithelial (RPE) cells and extracellular vesicle (EV) formation/release are related through the lysosomal and exocytotic pathways that process and eliminate intracellular material, including mitochondrial fragments. We propose that RPE cells with impaired mitochondria will release EVs containing mitochondrial miRNAs that reflect the diminished capacity of mitochondria within these cells. METHODS: We screened ARPE-19 cells for miRNAs that localize to the mitochondria, exhibit biological activity, and are present in EVs released by both untreated cells and cells treated with rotenone to induce mitochondrial injury. EVs were characterized by vesicle size, size distribution, presence of EV biomarkers: CD81, CD63, and syntenin-1, miRNA cargo, and number concentration of EVs released per cell. RESULTS: We found that miR-494-3p was enriched in ARPE-19 mitochondria. Knockdown of miR-494-3p in ARPE-19 cells decreased ATP production and mitochondrial membrane potential in a dose-dependent manner, and decreased basal oxygen consumption rate and maximal respiratory capacity. Increased number of EVs released per cell and elevated levels of miR-494-3p in EVs released from ARPE-19 cells treated with rotenone were also measured. CONCLUSIONS: ARPE-19 mitochondrial function is regulated by miR-494-3p. Elevated levels of miR-494-3p in EVs released by ARPE-19 cells indicate diminished capacity of the mitochondria within these cells. GENERAL SIGNIFICANCE: EV miR-494-3p is a potential biomarker for RPE mitochondrial dysfunction, which plays a central role in non-neovascular age-related macular degeneration, and may be a diagnostic biomarker for monitoring the spread of degeneration to neighboring RPE cells in the retina. SN - 1872-8006 UR - https://www.unboundmedicine.com/medline/citation/32240720/Release_of_extracellular_vesicle_miR_494_3p_by_ARPE_19_cells_with_impaired_mitochondria_ DB - PRIME DP - Unbound Medicine ER -