APR3 modulates oxidative stress and mitochondrial function in ARPE‐19 cells

Abstract

Impairment of retinal pigment epithelial (RPE) cells is considered a key contributor to the development of age-related macular degeneration. Apoptosis-related protein 3 (APR3) was recently discovered after treatment with all- trans retinoic acid, a pivotal molecule in RPE cells. However, the function of APR3 remains poorly understood. In the present study, we found that APR3 could interact with nuclear factor (erythroid-derived 2)-like 2, which is a regulator of phase II enzymes, and that knockdown of APR3 promoted nuclear factor (erythroid-derived 2)-like 2 nuclear translocation and activated expression of phase II enzymes, which was accompanied by improved redox status and mitochondrial activity. Overexpression of APR3 revealed its mitochondrial localization and induced a robust production of reactive oxygen species that was accompanied by impaired mitochondrial oxygen consumption, complex activity, and lower ATP content, resulting in significant changes in mitochondrial structure, which may contribute to cell apoptosis. High doses of all- trans retinoic acid treatment were found to significantly induce APR3 expression, increase reactive oxygen species levels, and decrease ATP content, which were abolished by knockdown of APR3. These results indicate that APR3 plays a vital role in regulating redox status and mitochondrial activity and thus suggest APR3 might be a potential novel target for study of treatment of age-related macular degeneration.-Li, Y., Zou, X., Gao, J., Cao, K., Feng, Z., Liu, J. APR3 modulates oxidative stress and mitochondrial function in ARPE-19 cells.

S-Allylmercapro-N-Acetylcysteine Attenuates the Oxidation-Induced Lens Opacification and Retinal Pigment Epithelial Cell Death In Vitro

“…The ARPE-19 human RPE cell line was purchased from the American Type Culture Collection (Manassas, VA, USA). ARPE-19 cells are an accepted cell model for the study of oxidative stress and anti-oxidants effects on RPE cells [23,24,25,26]. Cells were cultured in DMEM/F-12 (1:1) supplemented with 10% heat-inactivated Fetal Calf Serum, 2 mM l -Glutamine, 100 U/ml Penicillin, 100 μg/mL Streptomycin and 50 μg/mL Neomycin and passages 3 to 7 were used.…”

Functional characterization of all‐trans retinoic acid‐induced differentiation factor (ATRAID)

“…Shortly thereafter, it was recognized in a cDNA library derived from CD34+ hematopoietic stem/progenitor cells (HSPCs) and named HSPC013 [ 22 ]. Various isoforms of ATRAID have been used in previous studies, most often Iso B [ 5 , 6 , 7 , 8 , 10 , 23 , 24 ]. In NCBI, three RefSeq transcripts of ATRAID can be found, NM_016085 (Iso A), NM_080592 (Iso B), and NM_001170795 (Iso C).…”

“…In transfection experiments with N‐terminal‐tagged ATRAID Iso B, Li and coworkers observed that ATRAID interacted with NRF2 and KEAP1 in the mitochondria of ARPE‐19 cells [ 10 ]. In transduced cell lines expressing C‐terminal Myc‐Flag‐tagged ATRAID Iso C, we did not observe any colocalization between ATRAID and the mitochondrial protein TOMM20 (Fig.…”

“…All‐trans retinoic acid‐induced differentiation factor has been observed in mitochondria in transfected arising retinal pigment epithelial cells 19 (ARPE‐19) cells, involved in regulating mitochondrial phase II enzymes and the cellular redox status by binding to the complex of nuclear factor erythroid 2–related factor 2 (NRF2) and Kelch‐like ECH‐associated protein 1 (KEAP1) [ 10 ]. Han and coworkers reported increased the expression of ATRAID in RPE cells from aged mice.…”