Dataset Information

Proteomic analysis of ARPE-19 cells exposed to chronic H2O2 treatment

ABSTRACT: We performed a proteomic analysis of a retinal pigment epithelium cell line (ARPE-19) exposed to long-term sublethal levels of H2O2 to mimic low-level pro-oxidative insult in age-related macular degeneration (AMD). using a wide range of biochemical analyses and cell culture techniques, we highlighted changes in energy metabolism, extracellular matrix organization and inflammation-related processes, which may help in understanding the molecular pathogenesis of AMD.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Retinal Pigment Epithelial Cell, Cell Culture

DISEASE(S): Age Related Macular Degeneration

SUBMITTER: Davide Chiasserini

LAB HEAD: Davide Chiasserini

PROVIDER: PXD022545 | Pride | 2021-01-04

REPOSITORIES: Pride

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Publications

Crosstalk between Long-Term Sublethal Oxidative Stress and Detrimental Inflammation as Potential Drivers for Age-Related Retinal Degeneration.

Macchioni Lara L Chiasserini Davide D Mezzasoma Letizia L Davidescu Magdalena M Orvietani Pier Luigi PL Fettucciari Katia K Salviati Leonardo L Cellini Barbara B Bellezza Ilaria I

Antioxidants (Basel, Switzerland) 20201229 1

Age-related retinal degenerations, including age-related macular degeneration (AMD), are caused by the loss of retinal pigmented epithelial (RPE) cells and photoreceptors. The pathogenesis of AMD, deeply linked to the aging process, also involves oxidative stress and inflammatory responses. However, the molecular mechanisms contributing to the shift from healthy aging to AMD are still poorly understood. Since RPE cells in the retina are chronically exposed to a pro-oxidant microenvironment throu ...[more]

PMID: 33383836

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Project description:Background: Age-related macular degeneration (AMD) is caused by the degeneration of the macular photoreceptors. This is preceded by development of subretinal protein aggregates (drusen) and degeneration of the macular retinal pigment epithelium (RPE). The underlying pathogenesis remains unknown, but the immune system is suspected to play a key role in it. Aging of the immune system, immunosenescence, includes changes in T cell sub-populations and results in low-level chronic inflammation. Because AMD exclusively occurs in patients over 55 years of age, we hypothesize that aging of the T cell compartment may be implicated in AMD pathogenesis. Methodology and principal findings: Using an in vitro co-culture system, we investigated the effects of activated T cells on a human RPE cell line (ARPE-19). Differential gene expression in the RPE cells of complement factor genes was identified using microarrays, and selected gene transcripts from the alternative complement pathway were validated by q-RT-PCR. Protein expression was determined by ELISA and immunoblotting. Co-culture with activated T cells increased RPE mRNA and protein expression of complement component C3, factors B, H, H-like 1, CD46, CD59, and clusterin, in a dose-dependent manner. Conclusions: RPE cells responded to inflammatory assault caused by exposure to T cell-derived cytokines by upregulating expression of many complement factors from the alternative pathway. This may have implications for RPE ability to deal with complement attack, and opsonization and removal of debris from the RPE-choroidal interface. We speculate that regulation of the complement system in response to inflammatory assault may play a vital role in RPE pathology and drusen biogenesis. Experiment Overall Design: 10 samples investigating variations of co-cultures of RPE cells and T-cells. Variable parameters include the cell type, the co-culturing of the counter-part cell type and the orientation of the system (apical vs basolateral).

Dataset Information

Proteomic analysis of ARPE-19 cells exposed to chronic H2O2 treatment

Publications

Crosstalk between Long-Term Sublethal Oxidative Stress and Detrimental Inflammation as Potential Drivers for Age-Related Retinal Degeneration.

Similar Datasets

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