bims-mideyd 2023-11-19 papers

Cell Mol Biol (Noisy-le-grand). 2023 Oct 31. 69(10): 17-22

MicroRNA-626 inhibits mTOR pathways activity of retinal pigment epithelial cells by targeting SLC7A5 in human ARPE-19 Cells.

Çilem Ercan, Ahmet Elbay, Elif Sibel Aslan, Fahri Akbaş, Hakan Ozdemir, Nehir Ozdemir Ozgentürk.

Recent studies have shown that miRNAs are associated with the pathological process involved in age-related macular degeneration (AMD). However, the microRNA-mediated post-transcriptional regulation in human retinal pigment epithelium (RPE) cells has not been adequately investigated. We investigated how miR-626 inhibits mTOR activity pathways and pathway-related genes in retinal pigment epithelial cells by targeting the solute carrier family seven-member 5 (SLC7A5) in ARPE19 cells. We transfected mir-626 mimic, mir-626 inhibitör and siRNA in human retinal pigment epithelial cell line was examined using RT-PCR and western blot, respectively. We knocked down mir-626 levels and overexpression by mir-626-siRNA transfection of human RPE cell lines, and using an MTT assay, we assessed the role of SLC7A5 on RPE cell proliferation. We additionally measured the expression of mTOR, Akt1, caspase 3, Bax, SLC17A7, SLC17A8, Creb1, Pten, HIF1A, HIFI. The findings demonstrate that mir-626 inhibits SLC7A5 gene expression and proliferation of ARPE-19 cells. Short interfering RNA (siRNA) mediated suppression of SLC7A5, a predicted target of mir-626, has the same effect on ARPE-19 cells. We identified how miR-626 causes apoptosis and macula degeneration in RPE cells by targeting SLC7A5 through the mTOR signaling pathway. miR-626 was an essential regulator of the expression of the Slc7a5 gene. Importantly, we determined that miR-626 is essential to play a role in AMD. This research project shows that SLC7A5 is a direct target of mir-626 in ARPE-19 cells for the first time.

DOI: https://doi.org/10.14715/cmb/2023.69.10.3

Invest Ophthalmol Vis Sci. 2023 Nov 01. 64(14): 18

Endoplasmic Reticulum Stress Disrupts Mitochondrial Bioenergetics, Dynamics and Causes Corneal Endothelial Cell Apoptosis.

Saba Qureshi, Stephanie Lee, William Steidl, Lukas Ritzer, Michael Parise, Ananya Chaubal, Varun Kumar.

Purpose: Endoplasmic reticulum (ER) and mitochondrial stress are independently associated with corneal endothelial cell (CEnC) loss in many corneal diseases, including Fuchs' endothelial corneal dystrophy (FECD). However, the role of ER stress in mitochondrial dysfunction contributing to CEnC apoptosis is unknown. The purpose of this study is to explore the crosstalk between ER and mitochondrial stress in CEnC.
Methods: Human corneal endothelial cell line (HCEnC-21T) and human corneal endothelial tissues were treated with ER stressor tunicamycin. ER stress-reducing chemical 4-phenyl butyric acid (4-PBA) was used in HCEnC-21T after tunicamycin. Fuchs' corneal endothelial cell line (F35T) was used to determine differential activation of ER stress with respect to HCEnC-21T at the baseline. ER stress, mitochondrial-mediated intrinsic apoptotic, mitochondrial fission, and fusion proteins were determined using immunoblotting and immunohistochemistry. Mitochondrial bioenergetics were assessed by mitochondrial membrane potential (MMP) loss and ATP production at 48 hours after tunicamycin. Mitochondria dynamics (shape, area, perimeter) were also analyzed at 24 hours using transmission electron microscopy.
Results: Treatment of HCEnC-21T cell line with tunicamycin activated three ER stress pathways (PERK-eIF2α-CHOP, IRE1α-XBP1, and ATF6), reduced cell viability, upregulated mitochondrial-mediated intrinsic apoptotic molecules (cleaved caspase 9, caspase 3, PARP, Bax, cytochrome C), downregulated anti-apoptotic Bcl-2 protein, initiated mitochondrial dysfunction by loss of MMP and lowering of ATP production, and caused mitochondrial swelling and fragmentation with increased expression of mitochondrial fission proteins (Fis1 and p-Drp1). Fuchs' CEnC (F35T) cell line also showed activation of the ER stress-related proteins (p-eIF2α, GRP78, CHOP, XBP1) compared to HCEnC-21T at the baseline. The 4-PBA ameliorated cell loss and reduced cleaved caspase 3 and 9, thereby rescuing tunicamycin-induced cell death but not mitochondrial bioenergetics in HCEnC-21T cell line.
Conclusions: Tunicamycin-induced ER stress disrupts mitochondrial bioenegetics, dynamics and contributes to the loss of CEnC viability. This novel study highlights the importance of ER-mitochondria crosstalk and its contribution to CEnCs apoptosis, seen in many corneal diseases, including FECD.

DOI: https://doi.org/10.1167/iovs.64.14.18

Eur J Ophthalmol. 2023 Nov 17. 11206721231214297

Current trends in targeting the oxidative stress in glaucoma (Review).

Raluca Eugenia Iorga, Andreea Dana Moraru, Dănuț Costin, Răzvana Sorina Munteanu-Dănulescu, Daniel Constantin Brănișteanu.

Glaucoma is a progressive optic neuropathy characterised by retinal ganglion cell degeneration and visual field loss. Glaucoma is considered to be the leading cause of blindness in the industrialised countries. Oxidative damage is an important pathogenic factor in glaucoma, which triggers trabecular meshwork (TM) degeneration, which then leads to intraocular hypertension. Neurodegenerative insults during glaucomatous neurodegeneration initiate an immune response to restore tissue homeostasis. However, the oxidative stress (OS) that develops during the pathogenic processes of glaucoma, along with the agerelated OS, plays a critical role in shifting the physiological equilibrium. In the TM from glaucoma donors, proinflammatory markers were found, which were induced by the activation of a stress response. Chronic changes in the composition of antioxidants found in aqueous humour may induce alterations in TM as well as in the optic nerve head cells. Highlighting the pathogenic role of reactive oxygen species (ROS) in glaucoma has implications in preventing this disease. Various clinical trials are available to test the efficacy of antioxidant drugs in glaucoma management. In this review, we discuss the OS as a therapeutic target, suggesting that the modulation of a pro-oxidant/antioxidant status might be a relevant target for glaucoma prevention and therapy.

Keywords: Glaucoma; neuroprotection; oxidative stress; reactive oxygen species; retinal ganglion cells; trabecular meshwork

DOI: https://doi.org/10.1177/11206721231214297

Exp Eye Res. 2023 Nov 14. pii: S0014-4835(23)00349-4. [Epub ahead of print] 109728

Regulated cell death pathways in the sodium iodate model: Insights and implications for AMD.

Mala Upadhyay, Vera L Bonilha.

The sodium iodate (NaIO3) model of increased oxidative stress recapitulates dry AMD features such as patchy RPE loss, secondary photoreceptors, and underlying choriocapillaris death, allowing longitudinal evaluation of the retinal structure. Due to the time- and dose-dependent degeneration observed in diverse animal models, this preclinical model has become one of the most studied models. The events leading to RPE cell death post- NaIO3 injection have been extensively studied, and here we have reviewed different modalities of cell death, including apoptosis, necroptosis, ferroptosis, and pyroptosis with a particular focus on findings associated with in vivo and in vitro NaIO3 studies on RPE cell death. Because the fundamental cause of vision loss in patients with dry AMD is the death of these same cells affected by NaIO3, studies using NaIO3 can provide valuable insights into RPE and photoreceptor cell death mechanisms and can help understand mechanisms behind RPE degeneration in AMD.

Keywords: AMD; Cell death; Photoreceptor; RPE; Retinal degeneration; Sodium iodate

DOI: https://doi.org/10.1016/j.exer.2023.109728