Oxidative Stress and Dysfunctional Intracellular Traffic Linked to an Unhealthy Diet Results in Impaired Cargo Transport in the Retinal Pigment Epithelium (RPE)

• Published in: Molecular nutrition & food research 2019-08, Vol.63 (15), p.e1800951-n/a
• Main Authors: Keeling, Eloise, Chatelet, David S., Johnston, David A., Page, Anton, Tumbarello, David A., Lotery, Andrew J., Ratnayaka, J. Arjuna
• Format: Article
• Language: English
• Subjects: Algorithms; Autophagosomes - metabolism; Autophagy; Biological Transport; cargo trafficking; Cargo transportation; Cell Line; Diet; Diet - adverse effects; Epithelium; Humans; Hydrogen peroxide; Hydrogen Peroxide - pharmacology; Immunofluorescence; Intracellular; Intracellular Space - metabolism; Lysosomal Membrane Proteins - metabolism; Lysosomal-Associated Membrane Protein 2 - metabolism; Lysosomes; Macrolides - pharmacology; Macular degeneration; Microtubule-Associated Proteins - metabolism; Nutrition; Oxidative stress; Oxidative Stress - drug effects; Phagocytosis; phagosome and autophagy‐lysosomal pathways; Phagosomes; rab GTP-Binding Proteins - metabolism; rab5 GTP-Binding Proteins - metabolism; rab7 GTP-Binding Proteins; Retina; Retinal Photoreceptor Cell Outer Segment - metabolism; Retinal pigment epithelium; Retinal Pigment Epithelium - metabolism; Retinal Pigment Epithelium - pathology; Vesicles
• ISSN: 1613-4125; 1613-4133
• DOI: 10.1002/mnfr.201800951

Scope Oxidative stress and dysregulated intracellular trafficking are associated with an unhealthy diet which underlies pathology. Here, these effects on photoreceptor outer segment (POS) trafficking in the retinal pigment epithelium (RPE), a major pathway of disease underlying irreversible sight‐loss, are studied. Methods and results POS trafficking is studied in ARPE‐19 cells using an algorithm‐based quantification of confocal‐immunofluorescence data supported by ultrastructural studies. It is shown that although POS are tightly regulated and trafficked via Rab5, Rab7 vesicles, LAMP1/2 lysosomes and LC3b‐autophagosomes, there is also a considerable degree of variation and flexibility in this process. Treatment with H2O2 and bafilomycin A1 reveals that oxidative stress and dysregulated autophagy target intracellular compartments and trafficking in strikingly different ways. These effects appear limited to POS‐containing vesicles, suggesting a cargo‐specific effect. Conclusion The findings offer insights into how RPE cells cope with stress, and how mechanisms influencing POS transport/degradation can have different outcomes in the senescent retina. These shed new light on cellular processes underlying retinopathies such as age‐related macular degeneration. The discoveries reveal how diet and nutrition can cause fundamental alterations at a cellular level, thus contributing to a better understanding of the diet‐disease axis. Oxidative stress and dysfunctional intracellular membrane trafficking are disease processes linked with an unhealthy diet. Here, it is demonstrated how these adversely affect photoreceptor outer segment trafficking in the Retinal Pigment Epithelium (RPE); a cell‐layer critical to vision. The discoveries provide new insights into how diet‐induced pathogenic mechanisms damage RPE cells which results in irreversible sight loss. .