Exosomal miR-4488 and miR-1273g-5p inhibit the epithelial-mesenchymal transition of transforming growth factor β2-mediated retinal pigment epithelial cells by targeting ATP-binding cassette A4

Abstract

Exosomal microRNAs (miRNAs) have been shown to be involved in the regulation of many disease progression, including proliferative vitreoretinopathy (PVR). However, the roles of exosomal miR-4488 and miR-1273g-5p in PVR progression have not been demonstrated. Transforming growth factor β2 (TGF-β2)-induced ARPE-19 cells were used to stimulate the epithelial-mesenchymal transition (EMT) of cells.Exosomes derived from TGF-β2-induced ARPE-19 cells were identified by transmission electron microscopy and nanoparticle tracking analysis. The expression levels of miR-4488, miR-1273g-5p and ATP-binding cassette A4 (ABCA4) were measured by quantitative real-time PCR. The promotion levels of exosomes markers, EMT markers, apoptosis markers and ABCA4 were determined by western blot analysis. The migration, invasion and apoptosis of cells were determined by transwell assay, wound healing assay and flow cytometry. Our data showed that miR-4488 and miR-1273g-5p were lowly expressed in TGF-β2-induced ARPE-19 cells.Overexpressed exosomal miR-4488 and miR-1273g-5p could inhibit the EMT, migration, invasion, and promote apoptosis in TGF-β2-induced ARPE-19 cells. In addition, ABCA4 was a target of miR-4488 and miR-1273g-5p. Overexpressed ABCA4 also could reverse the negatively regulation of exosomal miR-4488 and miR-1273g-5p on the EMT, migration, and invasion of TGF-β2-induced ARPE-19 cells. In conclusion, our data showed that exosomal miR-4488 and miR-1273g-5p could inhibit TGF-β2-stimulated EMT in ARPE-19 cells through targeting ABCA4.

A Microfluidic Platform to Monitor Real-Time Effects of Extracellular Vesicle Exchange between Co-Cultured Cells across Selectively Permeable Barriers

“…Matrigel barriers and Transwell filter systems have been utilized previously, both to collect secreted EVs that have selectively diffused through the pores and accumulated in the supernatant [ 21 ], and to use pre-isolated EVs for cell migration studies [ 37 ]. Transwell systems have also been extensively used with or without supplementary Matrigel layers to investigate the role of certain exosome populations in cell migration [ 38 , 39 , 40 , 41 , 42 ]. Our microfluidic system circumvents the EV isolation process required for the aforementioned Transwell exosome functional strategies.…”

Puerarin suppresses hypoxia-induced vascular endothelial growth factor upregulation in human retinal pigmented epithelial cells by blocking JAK2/STAT3 pathway

“…Human RPE cells (ARPE-19 and D407 cells) were purchased from the Bacterial Collection Center of Wuhan University. These cells were cultured in a DMEM medium (Thermo fisher) that was supplemented with 1% non-essential amino acids, 29 mM sodium bicarbonate, 10% FBS, 20 mM HEPES, 100 μg/ml streptomycin and 100 U/ml penicillin (Sigma Aldrich) [ 20 ]. Cells were cultured at 37°C, 5% CO 2 humidity condition.…”

“…One-way ANOVA with post-hoc Tukey test was used to determine the significant difference among multiple groups (>2 groups). P < 0.05 was considered a significant difference [ 20 ].…”

Salivary Exosomes in Health and Disease: Future Prospects in the Eye

“…In addition to non-ocular diseases, exosomes have been implicated in the development and/or progression of several ocular diseases [ 256 , 257 , 258 , 259 ]. The subsequent sections will highlight the roles of exosomes in diabetic retinopathy, corneal disease, age-related macular degeneration, uveal disease, and glaucoma.…”

“…Notably, miR-543 was found in the EMT-induced RPE exosomes and significantly induced the EMT of the recipient RPE cells, highlighting the role of exosomes in triggering PVR via EMT induction [ 341 ]. Interestingly, exosomal miR-4488 and miR-1273g-5p have been reported to inhibit the TGF-β2-stimulated EMT in ARPE-19 cells, by downregulating the ATP-binding cassette A4 (ABCA4) [ 258 ], which has been reported to be elevated in PVR tissue [ 342 ]. Overexpressed ABCA4 can counteract the inhibitory effect of miR-4488 and miR-1273g-5p on the proliferation, migration, and invasion of TGF-β2-stimulated ARPE-19 cells, suggesting that ABCA4-depleting therapies can slow PVR progression [ 258 ].…”