Quantitative proteomic analysis of aqueous humor from patients with drusen and reticular pseudodrusen in age-related macular degeneration

Autor: Kyu Hyung Park, Daehan Lim, Je Hyun Baek, Jonghyun Lee, Hyoik Jang, Jae Byoung Chae, Hye Won Chung
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Male
Proteomics
Proteome
genetic structures
Cathepsin D
Retinal Pigment Epithelium
Mass Spectrometry
chemistry.chemical_compound
0302 clinical medicine
lcsh:Ophthalmology
Electric Impedance
Fluorescent Antibody Technique
Indirect
Cells
Cultured
medicine.diagnostic_test
General Medicine
Blot
Phenotype
Female
Tomography
Optical Coherence
Extracellular matrix organization
Research Article
Reticular pseudodrusen
Blotting
Western
Complement
Enzyme-Linked Immunosorbent Assay
Retinal Drusen
Drusen
Immunofluorescence
Aqueous Humor
03 medical and health sciences
Geographic Atrophy
medicine
Humans
Eye Proteins
Aged
Aldehydes
business.industry
Age-related macular degeneration
Retinal
Macular degeneration
medicine.disease
Molecular biology
eye diseases
Ophthalmology
Oxidative Stress
030104 developmental biology
chemistry
lcsh:RE1-994
030221 ophthalmology & optometry
SWATH-MS
sense organs
business
Biomarkers
Zdroj: BMC Ophthalmology, Vol 18, Iss 1, Pp 1-13 (2018)
BMC Ophthalmology
ISSN: 1471-2415
Popis: Background To identify novel biomarkers related to the pathogenesis of dry age-related macular degeneration (AMD), we adopted a human retinal pigment epithelial (RPE) cell culture model that mimics some features of dry AMD including the accumulation of intra- and sub-RPE deposits. Then, we investigated the aqueous humor (AH) proteome using a data-independent acquisition method (sequential window acquisition of all theoretical fragment ion mass spectrometry) for dry AMD patients and controls. Methods After uniformly pigmented polarized monolayers of human fetal primary RPE (hfRPE) cells were established, the cells were exposed to 4-hydroxy-2-nonenal (4-HNE), followed by Western blotting, immunofluorescence analysis and ELISA of cells or conditioned media for several proteins of interest. Data-dependent acquisition for identification of the AH proteome and SWATH-based mass spectrometry were performed for 11 dry AMD patients according to their phenotypes (including soft drusen and reticular pseudodrusen [RPD]) and 2 controls (3 groups). Results Increased intra- and sub-RPE deposits were observed in 4-HNE-treated hfRPE cells compared with control cultures based on APOA1, cathepsin D, and clusterin immunoreactivity. Additionally, the differential abundance of proteins in apical and basal chambers with or without 4-HNE treatment confirmed the polarized secretion of proteins from hfRPE cells. A total of 119 proteins were quantified in dry AMD patients and controls by SWATH-MS. Sixty-five proteins exhibited significantly altered abundance among the three groups. A two-dimensional principal component analysis plot was generated to identify typical proteins related to the pathogenesis of dry AMD. Among the identified proteins, eight proteins, including APOA1, CFHR2, and CLUS, were previously considered major components or regulators of drusen. Three proteins (SERPINA4, LUM, and KERA proteins) have not been previously described as components of drusen or as being related to dry AMD. Interestingly, the LUM and KERA proteins, which are related to extracellular matrix organization, were upregulated in both RPD and soft drusen. Conclusions Differential protein expression in the AH between patients with drusen and RPD was quantified using SWATH-MS in the present study. Detailed proteomic analyses of dry AMD patients might provide insights into the in vivo biology of drusen and RPD. Electronic supplementary material The online version of this article (10.1186/s12886-018-0941-9) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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