Dataset Information

Quantitative phosphoproteomic analysis of ARPE-19 cells in response to LPS

ABSTRACT: Overwhelming and persistent inflammation of retinal pigment epithelium (RPE) induces destructive changes in retinal environment by invoking immune activation. In this study, we aimed to investigate RPE-specific biological and metabolic response against intense inflammation, and identify molecular characteristics determining pathological progression. Here, we performed quantitative analyses of phosphoproteome of lipopolysaccharide (LPS)-stimulated human derived RPE cell line ARPE-19 using the latest isobaric TMT labeling approach coupled with high-resolution mass spectrometry.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Retinal Pigment Epithelium Cell Line, Epithelial Cell

SUBMITTER: Dohyun Han

LAB HEAD: Dohyun Han

PROVIDER: PXD018260 | Pride | 2021-09-09

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20191212_ARPE19_LPS_TMT10_TI02_F1.raw	Raw
	20191212_ARPE19_LPS_TMT10_TI02_F10.raw	Raw
	20191212_ARPE19_LPS_TMT10_TI02_F11.raw	Raw
	20191212_ARPE19_LPS_TMT10_TI02_F12.raw	Raw
	20191212_ARPE19_LPS_TMT10_TI02_F2.raw	Raw

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Publications

A Comprehensive Proteomic and Phosphoproteomic Analysis of Retinal Pigment Epithelium Reveals Multiple Pathway Alterations in Response to the Inflammatory Stimuli.

Song Juha J Han Dohyun D Lee Heonyi H Kim Da Jung DJ Cho Joo-Youn JY Park Jae-Hak JH Seok Seung Hyeok SH

International journal of molecular sciences 20200425 9

Overwhelming and persistent inflammation of retinal pigment epithelium (RPE) induces destructive changes in the retinal environment. However, the precise mechanisms remain unclear. In this study, we aimed to investigate RPE-specific biological and metabolic responses against intense inflammation and identify the molecular characteristics determining pathological progression. We performed quantitative analyses of the proteome and phosphoproteome of the human-derived RPE cell line ARPE-19 after tr ...[more]

PMID: 32344885

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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).