scholarly journals A Splice Variant in SLC16A8 Gene Leads to Lactate Transport Deficit in Human iPS Cell-Derived Retinal Pigment Epithelial Cells

Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 179
Author(s):  
Laurence Klipfel ◽  
Marie Cordonnier ◽  
Léa Thiébault ◽  
Emmanuelle Clérin ◽  
Frédéric Blond ◽  
...  
Keyword(s):  
Genome Wide Association Study ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Lactate Transport ◽  
Ips Cell ◽  
Risk Alleles ◽  
Age Related ◽  
A Genome

Age-related macular degeneration (AMD) is a blinding disease for which most of the patients remain untreatable. Since the disease affects the macula at the center of the retina, a structure specific to the primate lineage, rodent models to study the pathophysiology of AMD and to develop therapies are very limited. Consequently, our understanding relies mostly on genetic studies highlighting risk alleles at many loci. We are studying the possible implication of a metabolic imbalance associated with risk alleles within the SLC16A8 gene that encodes for a retinal pigment epithelium (RPE)-specific lactate transporter MCT3 and its consequences for vision. As a first approach, we report here the deficit in transepithelial lactate transport of a rare SLC16A8 allele identified during a genome-wide association study. We produced induced pluripotent stem cells (iPSCs) from the unique patient in our cohort that carries two copies of this allele. After in vitro differentiation of the iPSCs into RPE cells and their characterization, we demonstrate that the rare allele results in the retention of intron 2 of the SLC16A8 gene leading to the absence of MCT3 protein. We show using a biochemical assay that these cells have a deficit in transepithelial lactate transport.

scholarly journals Human iPS cell derived RPE strips for secure delivery of graft cells at a target place with minimal surgical invasion

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mitsuhiro Nishida ◽  
Yuji Tanaka ◽  
Yo Tanaka ◽  
Satoshi Amaya ◽  
Nobuyuki Tanaka ◽  
...  
Keyword(s):  
Target Location ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Cell Sheet ◽  
Invasive Procedure ◽  
Age Related Macular Degeneration ◽  
Target Area ◽  
Ips Cell ◽  
Age Related

AbstractSeveral clinical studies have been conducted into the practicality and safety of regenerative therapy using hESC/iPSC-retinal pigment epithelium (RPE) as a treatment for the diseases including age-related macular degeneration. These studies used either suspensions of RPE cells or an RPE cell sheet. The cells can be injected using a minimally invasive procedure but the delivery of an intended number of cells at an exact target location is difficult; cell sheets take a longer time to prepare, and the surgical procedure is invasive but can be placed at the target area. In the research reported here, we combined the advantages of the two approaches by producing a quickly formed hiPSC-RPE strip in as short as 2 days. The strip readily expanded into a monolayer sheet on the plate, and after transplantation in nude rats, it showed a potency to partly expand with the correct apical/basal polarity in vivo, although limited in expansion area in the presence of healthy host RPE. The strip could be injected into a target area in animal eyes using a 24G canula tip.

scholarly journals Hypoxia-induced metabolic stress in retinal pigment epithelial cells is sufficient to induce photoreceptor degeneration

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Toshihide Kurihara ◽  
Peter D Westenskow ◽  
Marin L Gantner ◽  
Yoshihiko Usui ◽  
Andrew Schultz ◽  
...  
Keyword(s):  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Metabolic Stress ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Photoreceptor Degeneration ◽  
Rpe Cells ◽  
Glucose And Lipid Metabolism ◽  
Age Related ◽  
Cellular Stresses

Photoreceptors are the most numerous and metabolically demanding cells in the retina. Their primary nutrient source is the choriocapillaris, and both the choriocapillaris and photoreceptors require trophic and functional support from retinal pigment epithelium (RPE) cells. Defects in RPE, photoreceptors, and the choriocapillaris are characteristic of age-related macular degeneration (AMD), a common vision-threatening disease. RPE dysfunction or death is a primary event in AMD, but the combination(s) of cellular stresses that affect the function and survival of RPE are incompletely understood. Here, using mouse models in which hypoxia can be genetically triggered in RPE, we show that hypoxia-induced metabolic stress alone leads to photoreceptor atrophy. Glucose and lipid metabolism are radically altered in hypoxic RPE cells; these changes impact nutrient availability for the sensory retina and promote progressive photoreceptor degeneration. Understanding the molecular pathways that control these responses may provide important clues about AMD pathogenesis and inform future therapies.

ATTACHMENT AND PROLIFERATION OF RETINAL PIGMENT EPITHELIAL CELLS ON SMALL INTESTINE SUBMUCOSA POWDER IMPREGNATED POLY(L-LACTIDE-CO-GLYCOLIDE) FILM

2011 ◽  
Vol 23 (02) ◽  
pp. 119-126 ◽  
Author(s):  
Eun Hye Jo ◽  
Ga Young Lee ◽  
Su Jin Cho ◽  
Hanna Yoo ◽  
On You Kim ◽  
...  
Keyword(s):  
Small Intestine ◽  
Messenger Rna ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Cellular Adhesion ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Immunocytochemical Staining ◽  
Small Intestine Submucosa ◽  
Age Related

The retinal pigment epithelium (RPE) closely interacts with photoreceptors in the maintenance of visual function. The native RPEs exist as a monolayer structure and have a mottled brown color due to the presence of melanin and other pigments including lipofuscin granules, which accumulate with age. In age-related macular degeneration (AMD), RPE's dysfunction and changes in Bruch's membrane occur. Thus, small intestine submucosa/poly(lactic-co-glycolic acid) (SIS/PLGA) film is a biomimetic transplant consisting of a layer of healthy RPE cells cultured on a support membrane. The goals of this study were to evaluate the effects of attachment and proliferation of RPEs on SIS/PLGA films. Porcine SIS is an acellular tissue and widely used as a biomaterial without immunorejection responses, whereas PLGA is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. We fabricated SIS/PLGA films using 20 wt% of SIS. We measured MTT to confirm cellular adhesion of cell number attached on film at 1, 3, 5, and 7 days. Morphology of cellular adhesion on films was confirmed by scanning electron microscopy at 1, 3, and 7 days. Further, reverse transcription polymerase chain reaction (RT-PCR) was conducted to confirm messenger RNA expression of RPE65 as RPE's marker and expression of cytokeratin, and RPE65 were determined by AEC immunocytochemical staining. These results suggest that SIS provides suitable surface to RPEs.

scholarly journals Mitophagy in the Retinal Pigment Epithelium of Dry Age-Related Macular Degeneration Investigated in the NFE2L2/PGC-1α-/- Mouse Model

2020 ◽  
Vol 21 (6) ◽  
pp. 1976 ◽  
Author(s):  
Iswariyaraja Sridevi Gurubaran ◽  
Johanna Viiri ◽  
Ali Koskela ◽  
Juha M.T. Hyttinen ◽  
Jussi J. Paterno ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Mouse Model ◽  
Macular Degeneration ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Double Knockout ◽  
Rpe Cells ◽  
Age Related

Increased oxidative stress and mitochondrial damage are observed in protein aggregation diseases, such as age-related macular degeneration (AMD). We have recently reported elevated levels of oxidative stress markers, damaged mitochondria, accumulating lysosomal lipofuscin and extracellular drusen-like structures in the retinal pigment epithelial cells (RPE) of the dry AMD-resembling NFE2L2/PGC1α double knockout (dKO) mouse model. Here, we provide evidence of a disturbance in the autolysosomal machinery handling mitochondrial clearance in the RPE cells of one-year-old NFE2L2/PGC1α-deficient mice. Confocal immunohistochemical analysis revealed an upregulation of autophagosome marker microtubule-associated proteins 1A/1B light chain 3B (LC3B) as well as numerous mitophagy markers, such as PTE-induced putative kinase 1 (PINK1) and E3 ubiquitin ligase (PARKIN) together with damaged mitochondria. However, we detected no evidence of increased autolysosome formation in transmission electron micrographs or of colocalization of lysosomal marker LAMP2 (lysosome-associated membrane protein 2) and the mitochondrial marker ATP synthase β in confocal micrographs. Interestingly, we observed an upregulation of late autolysosomal fusion Ras-related protein (Rab7) in the perinuclear space of RPE cells together with autofluorescence aggregates. Our results reveal that there is at least a relative decrease of mitophagy in the RPE cells of NFE2L2/PGC1α dKO mice. This further supports the hypothesis that mitophagy is a putative therapy target in AMD-like pathology.

scholarly journals Overview of how N32 and N34 elovanoids sustain sight by protecting retinal pigment epithelial cells and photoreceptors

2020 ◽  
pp. jlr.TR120001137 ◽  
Author(s):  
Nicolas G. Bazan
Keyword(s):  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Acyl Chain ◽  
Hexaenoic Acid ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Low Grade ◽  
Omega 3 ◽  
Age Related ◽  
And Function

The essential fatty acid DHA (22:6, omega-3 or n-3) is enriched in and required for the membrane biogenesis and function of photoreceptor cells (PRC), synapses, mitochondria, etc. of the CNS. PRC DHA becomes an acyl chain at the sn-2 of phosphatidylcholine (PC), amounting to more than 50% of the PRC outer segment phospholipids, where phototransduction takes place. Very long chain PUFAs (VLC-PUFAs,n-3, ≥ 28 carbons) are at the sn-1 of this PC molecular species and interact with rhodopsin. PRC shed their tips (DHA-rich membrane disks) daily, which in turn are phagocytized by the retinal pigment epithelium (RPE), where DHA is recycled back to PRC inner segments to be used for the biogenesis of new photoreceptor membranes. Here, we review the structures and stereochemistry of novel elovanoid (ELV)-N32 and ELV-N34 to be ELV-N32: (14Z,17Z,20R,21E,23E,25Z,27S,29Z)-20,27-dihydroxydo-triaconta-14,17,21,23,25,29-hexaenoic acid; ELV-N34: (16Z,19Z,22R,23E,25E,27Z,29S,31Z)-22,29-dihydroxytetra-triaconta-16,19,23,25,27,31-hexaenoic acid. ELVs are low-abundance, high-potency, protective mediators. Their bioactivity includes enhancing of anti-apoptotic and pro-survival protein expression with concomitant downregulation of pro-apoptotic proteins when RPE is confronted with uncompensated oxidative stress (UOS). ELVs also target PRC/RPE senescence gene programming, the senescence secretory phenotype in the interphotoreceptor matrix (IPM), as well as inflammaging (chronic, sterile, low-grade inflammation). An important lesson on neuroprotection is highlighted by the ELV mediators that target the terminally differentiated PRC and RPE, sustaining a beautifully synchronized renewal process. The role of ELVs in PRC and RPE viability and function uncovers insights on disease mechanisms and the development of therapeutics for age-related macular degeneration (AMD), Alzheimer’s disease (AD), and other pathologies.

scholarly journals The CD36 Ligand-Promoted Autophagy Protects Retinal Pigment Epithelial Cells from Oxidative Stress

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Marie-France Dorion ◽  
Mukandila Mulumba ◽  
Shuya Kasai ◽  
Ken Itoh ◽  
William D. Lubell ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Autophagic Flux ◽  
Cytoprotective Effect ◽  
Rpe Cells ◽  
Age Related ◽  
Cluster Of Differentiation

The retinal pigment epithelium (RPE) performs many functions that maintain photoreceptor health. Oxidative damage to the RPE is a critical component in the pathogenesis of eye diseases such as age-related macular degeneration (AMD). Ligands of the cluster of differentiation 36 (CD36) have previously preserved photoreceptor integrity in mouse models of AMD. The cytoprotective effect of the CD36 ligand MPE-001 on RPE cells has now been elucidated employing a model of oxidative stress. Sodium iodate (NaIO3) induced formation of reactive oxygen species and apoptosis in human RPE cells, which were decreased by MPE-001 without affecting antioxidant enzyme transcription. Immunoblotting and immunostaining assays showed a restorative effect of MPE-001 on the autophagic flux disrupted by NaIO3, which was associated with an increase in syntaxin 17-positive mature autophagosomes. The cytoprotective effect of MPE-001 was completely abolished by the autophagy inhibitors wortmannin and bafilomycin A1. In conclusion, we report for the first time an autophagy-dependent protection of RPE cells from oxidative stress by a CD36 ligand.

scholarly journals Gastrodin represses hydrogen peroxide-induced oxidative stress in retinal pigment epithelial cells through p38MAPK/iNOS pathway

2021 ◽  
Vol 13 (4) ◽  
pp. 24-30
Author(s):  
Xingli Zhou ◽  
Ximing Zhao
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Cell Viability ◽  
Cell Apoptosis ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
B Cell Lymphoma ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Age Related

Elevated reactive oxygen species (ROS) induce oxidative damage in retinal pigment epithelium (RPE) and con-tribute to the development of age-related macular degeneration (AMD). Gastrodin plays an antioxidant role in distinct diseases, such as epilepsy, cerebral ischemia, Alzheimer’s disease, and cardiovascular diseases. However, the function of gastrodin in AMD remains unclear. Human RPE (ARPE-19) cells were incubated with 300 μM hydrogen peroxide (H2O2) for 24 hours. The results showed that H2O2 decreased cell viability and promoted the cell apoptosis of ARPE-19 cells. H2O2-induced ARPE-19 cells were then treated with different concentrations of gastrodin. Gastrodin increased cell viability of H2O2-induced ARPE-19 cells, suppressed the cell apoptosis of H2O2-induced ARPE-19 cells with reduced B-cell lymphoma (Bcl)-2 like protein (Bax), and enhanced Bcl-2. The levels of ROS were enhanced, malondialdehyde (MDA) was up-regulated, and superoxide dismutase (SOD) and glutathione (GSH) were down-regulated in H2O2-induced ARPE-19 cells. However, gastrodin reduced the lev-els of ROS and MDA and elevated SOD and GSH in H2O2-induced ARPE-19 cells. Furthermore, H2O2-induced increase of inducible nitric oxide synthase (iNOS) and p-p38 proteins in ARPE-19 was reversed by gastrodin. In conclusion, gastrodin exerted antiapoptotic and antioxidant capacities to protect against H2O2-induced oxidative stress in RPE, thereby acting as a potential agent for managing AMD.

scholarly journals Comparative study of oxidant properties of oxidized and unoxidized bisretinoids of lipofuscine granules in the retinal pigment epithelium of human eye

2019 ◽  
pp. 66-71
Author(s):  
М.А. Яковлева ◽  
Н.Л. Сакина ◽  
И.Б. Кольчугина ◽  
П.М. Арбуханова ◽  
С.А. Борзенок ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Retinal Pigment Epithelium ◽  
Comparative Study ◽  
Degradation Products ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Age Related ◽  
Lipofuscin Granules

Актуальность. Недавно нами было показано, что при возрастной макулярной дегенерации сетчатки наблюдается повышенное содержание продуктов фотоокисления и фотодеградации бисретиноидов по сравнению с нормой. Поэтому на сегодняшний день вопрос о фототоксичности этих продуктов становится актуальным для решения проблемы поиска путей лечения и профилактики патологии. Цель. Провести сравнительное исследование фотосенсибилизирующего действия N-ретинилиден-N-ретинилэтаноламина (А2Е) и продуктов его фотоокисления и фотодеградации на индуцированную видимым светом пероксидацию липидов фоторецепторных мембран. Материалы и методы. При помощи метода высокоэффективной жидкостной хроматографии были получены отдельные фракции неокисленных и окисленных бисретиноидов в хлороформном экстракте липофусциновых гранул из ретинального пигментного эпителия кадаверных глаз. Результаты. Проведено сравнительное исследование фототоксических свойств неокисленных и окисленных бисретиноидов липофусциновых гранул из клеток ретинального пигментного эпителия глаза человека на пероксидацию липидов наружных сегментов фоторецепторных клеток. Выводы. Окисленные бисретиноиды липофусциновых гранул менее фототоксичны по сравнению с их неокисленными формами. Background. Recently we have shown that age-related macular degeneration is associated with higher than normal levels of bisretinoid photo-oxidation and photo-degradation products. Therefore, the issue of their phototoxicity currently becomes relevant for finding ways to treat and prevent this pathology. Aim. To conduct a comparative study of the photosensitizing effect of N-retinylidene-N-retinylethanolamine (A2E) and its photooxidation and photodegradation products on light-induced lipid peroxidation in photoreceptor membranes. Materials and methods. Using high-performance liquid chromatography fractions of unoxidized and oxidized bisretinoids were isolated in the chloroform extract of lipofuscin granules from the retinal pigment epithelium of cadaver eyes. Results. The study compared phototoxic effects of unoxidized and oxidized bisretinoids of lipofuscin granules from human retinal pigment epithelial cells on lipid peroxidation in rod outer segments. Conclusions. Oxidized bisretinoids of lipofuscin granules are less phototoxic compared to their unoxidized forms.

scholarly journals Madecassoside protects retinal pigment epithelial cells against hydrogen peroxide-induced oxidative stress and apoptosis through the activation of Nrf2/HO-1 pathway

2020 ◽  
Vol 40 (10) ◽  
Author(s):  
Jinzi Zhou ◽  
Fenghua Chen ◽  
Aimin Yan ◽  
Xiaobo Xia
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Triterpenoid Saponin ◽  
Sod Activity ◽  
Rpe Cells ◽  
Age Related

Abstract Age-related macular degeneration (AMD) is a progressive and degenerative ocular disease associated with oxidative stress. Madecassoside (MADE) is a major bioactive triterpenoid saponin that possesses antioxidative activity. However, the role of MADE in AMD has never been investigated. In the current study, we aimed to evaluate the protective effect of MADE on retinal pigment epithelium (RPE) cells under oxidative stress condition. We used hydrogen peroxide (H2O2) to induce oxidative damage in human RPE cells (ARPE-19 cells). Our results showed that H2O2-caused significant decrease in cell viability and increase in lactate dehydrogenase (LDH) release were dose-dependently attenuated by MADE. MADE treatment also attenuated H2O2-induced reactive oxygen species (ROS) and malondialdehyde (MDA) production in RPE cells. The reduced glutathione (GSH) level and superoxide dismutase (SOD) activity in H2O2-induced ARPE-19 cells were elevated after MADE treatment. MADE also suppressed caspase-3 activity and bax expression, as well as increased bcl-2 expression. Furthermore, H2O2-induced increase in expression levels of HO-1 and nuclear Nrf2 were enhanced by MADE treatment. Finally, knockdown of Nrf2 reversed the protective effects of MADE on H2O2-induced ARPE-19 cells. In conclusion, these findings demonstrated that MADE protected ARPE-19 cells from H2O2-induced oxidative stress and apoptosis by inducing the activation of Nrf2/HO-1 signaling pathway.

scholarly journals Piceatannol Protects Human Retinal Pigment Epithelial Cells against Hydrogen Peroxide Induced Oxidative Stress and Apoptosis through Modulating PI3K/Akt Signaling Pathway

Nutrients ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 1515 ◽  
Author(s):  
Yiming Hao ◽  
Jie Liu ◽  
Ziyuan Wang ◽  
Liangli (Lucy) Yu ◽  
Jing Wang
Keyword(s):  
Hydrogen Peroxide ◽  
Nuclear Translocation ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelium Cell ◽  
Retinal Pigment Epithelial Cells ◽  
Age Related Macular Degeneration ◽  
Ros Generation ◽  
Related Factor ◽  
Age Related

This study investigated the protective effect and the molecular mechanism of piceatannol on hydrogen peroxide (H2O2)-induced retinal pigment epithelium cell (ARPE-19) damage. Piceatannol treatment significantly inhibited H2O2-induced RPE cell death and reactive oxygen species (ROS) generation by 64.4% and 75.0%, respectively. Results of flow cytometry showed that H2O2-induced ARPE-19 cells apoptosis was ameliorated by piceatannol supplementation, along with decreased relative protein expressions of Bax/Bcl-2, Cleave-Caspase-3, and Cleave-PARP. Moreover, piceatannol treatment induced NF-E2-related factor 2 (Nrf2) signaling activation, which was evidenced by increased transcription of anti-oxidant genes, glutamate-cysteine ligase catalytic subunit (GCLc), SOD, and HO-1. Knockdown of Nrf2 through targeted siRNA alleviated piceatannol-mediated HO-1 transcription, and significantly abolished piceatannol-mediated cytoprotection. LY294002 (PI3K inhibitor) dramatically blocked piceatannol-mediated increasing of Nrf2 nuclear translocation, HO-1 expression, and cytoprotective activity, indicating the involvement of PI3K/Akt pathway in the cytoprotective effect of piceatannol. The results from this suggest the potential of piceatannol in reducing the risk of age-related macular degeneration.

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