Dataset Information

Mouse retinal pigmented epithelium (RPE)-LC/MS/MS

ABSTRACT: In this experiment, we compared the phosphorylated protein profile of Cryba1 cKO (RPE-specific) RPE tissue and that of the age-matched control samples.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

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

DISEASE(S): Age Related Macular Degeneration

SUBMITTER: Peng Shang

LAB HEAD: Debasish Sinha

PROVIDER: PXD026777 | Pride | 2021-09-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	Appendix2_JHU-Sinha-TMT10plex-Phos-peptidesV02-PRIDE.xlsx	Xlsx
	JHU-Sinha-TMT10plex-Phos1.raw	Raw
	JHU-Sinha-TMT10plex-Phos2.raw	Raw
	JHU-Sinha-TMT10plex-Phos3.raw	Raw
	JHU-Sinha-TMT10plex-Phos4.raw	Raw

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Publications

βA3/A1-crystallin regulates apical polarity and EGFR endocytosis in retinal pigmented epithelial cells.

Shang Peng P Stepicheva Nadezda N Teel Kenneth K McCauley Austin A Fitting Christopher Scott CS Hose Stacey S Grebe Rhonda R Yazdankhah Meysam M Ghosh Sayan S Liu Haitao H Strizhakova Anastasia A Weiss Joseph J Bhutto Imran A IA Lutty Gerard A GA Jayagopal Ashwath A Qian Jiang J Sahel José-Alain JA Samuel Zigler J J Handa James T JT Sergeev Yuri Y Rajala Raju V S RVS Watkins Simon S Sinha Debasish D

Communications biology 20210708 1

The retinal pigmented epithelium (RPE) is a monolayer of multifunctional cells located at the back of the eye. High membrane turnover and polarization, including formation of actin-based apical microvilli, are essential for RPE function and retinal health. Herein, we demonstrate an important role for βA3/A1-crystallin in RPE. βA3/A1-crystallin deficiency leads to clathrin-mediated epidermal growth factor receptor (EGFR) endocytosis abnormalities and actin network disruption at the apical side th ...[more]

PMID: 34239035

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Project description:Purpose: Age-related degeneration (AMD) is a major cause of blindness in developed countries. The molecular pathogenesis of early events in AMD is poorly understood. We investigated differential gene expression in samples of human retinal pigment epithelium (RPE)/choroid from early AMD and control maculas using exon-based arrays. Methods: Gene expression levels in nine early AMD and nine control human donor eyes were assessed using Affymetrix Human Exon ST 1.0 arrays. Two controls did not pass quality control and were removed. Differentially expressed genes were annotated using DAVID, and gene set enrichment analysis (GSEA) was performed on RPE-specific and endothelium-associated gene sets. CFH genotype was also assessed and differential expression was analyzed with respect to high AMD risk (YH/HH) and low AMD risk (YY) genotypes. Results: Seventy-five genes were identified as differentially expressed (raw p-value < 0.01; >50% fold change, mean log2 expression level in AMD or control M-bM-^IM-% median of all average gene expression values); however, no genes were significant (adj. p-value < 0.01) after correction for multiple hypothesis testing. Of 52 genes with decreased expression in AMD (fold change < 0.5; raw p-value < 0.01), 18 genes were identified by DAVID analysis as associated with vision or neurological processes. GSEA of RPE-associated and endothelium-associated genes revealed a significant decrease in genes typically expressed by endothelial cells in the early AMD group compared to controls, consistent with previous histologic and proteomic studies. Analysis with respect to CFH genotype indicated decreased expression of ADAMTS9 in eyes with high-risk genotypes (fold change = -2.61; raw p-value = 0.0008). Conclusions: GSEA results suggest that RPE transcripts are preserved or elevated in early AMD, concomitant with loss of endothelial cell marker expression. These results are consistent with the notion that choroidal endothelial cell dropout occurs early in the pathogenesis of AMD. Using AltAnalyze (ver. 2.0.7 beta), we analyzed nine early AMD and nine control eyes using Affymetrix Human Exon ST 1.0 arrays. Following initial processing in AltAnalyze, two control arrays were identified as potential outliers by tests implement in arrayQualityMetrics, a package for R.