Dataset Information

Proteomic and phosphoproteomic profiling of ARPE19 cells and Abca4-/-Rdh8-/- mice exposed to stress

ABSTRACT: To obtain an unbiased, thorough understanding of the biological pathways implicated in atRAL-induced cytotoxicity and PDE inhibitor-mediated cytoprotection, we performed a mass spectrometry-based quantitative proteomic analysis of ARPE19 cells, using stable-isotope labeling of amino acids in cell culture (SILAC). We identified differentially expressed proteins in ARPE19 cells exposed to stress (60 μM atRAL or 5 mM H2O2) in the absence or presence of PDE inhibitor (10 μM BAY 60-7550, rolipram, or BC11-38), relative to vehicle-treated controls. We also performed combined label-free mass spectrometry-based quantitative proteomic and phosphoproteomic analyses of retinas from the Abca4-/-Rdh8-/- double-knockout (dKO) light-sensitive mouse model of photoreceptor degeneration. First, we identified differentially expressed proteins in retinas exposed to bright-light stress (10,000 lux for 30 min) relative to unstressed controls. Next, we identified differentially expressed proteins in PDE inhibitor-treated mice (2 mg/kg intraperitoneal BAY 60-7550, rolipram, or BC11-38) exposed to stress relative to DMSO vehicle-treated controls. Altogether, these analyses provided insights into the proteomic and phosphoproteomic changes induced by stress and modulated by PDE-inhibitor therapy.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Retinal Pigment Epithelial Cell, Retina

DISEASE(S): Retinal Degeneration

SUBMITTER: Fangyuan Gao

LAB HEAD: Krzysztof Palczewski

PROVIDER: PXD035841 | Pride | 2024-01-26

REPOSITORIES: Pride

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Publications

Stress resilience-enhancing drugs preserve tissue structure and function in degenerating retina via phosphodiesterase inhibition.

Luu Jennings C JC Saadane Aicha A Leinonen Henri H Choi Elliot H EH Gao Fangyuan F Lewandowski Dominik D Halabi Maximilian M Sander Christopher L CL Wu Arum A Wang Jacob M JM Singh Rupesh R Gao Songqi S Lessieur Emma M EM Dong Zhiqian Z Palczewska Grazyna G Mullins Robert F RF Peachey Neal S NS Kiser Philip D PD Tabaka Marcin M Kern Timothy S TS Palczewski Krzysztof K

Proceedings of the National Academy of Sciences of the United States of America 20230501 19

Chronic, progressive retinal diseases, such as age-related macular degeneration (AMD), diabetic retinopathy, and retinitis pigmentosa, arise from genetic and environmental perturbations of cellular and tissue homeostasis. These disruptions accumulate with repeated exposures to stress over time, leading to progressive visual impairment and, in many cases, legal blindness. Despite decades of research, therapeutic options for the millions of patients suffering from these disorders remain severely l ...[more]

PMID: 37126699

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