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Quantitative proteomic analysis of polarized RPE secretome

ABSTRACT: To investigate the impact of serine protease HtrA1 on the polarized RPE secretome.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Chia-yi Chen

LAB HEAD: Oliver Schilling

PROVIDER: PXD007691 | Pride | 2018-04-09

REPOSITORIES: Pride

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Publications

N-Terminomics identifies HtrA1 cleavage of thrombospondin-1 with generation of a proangiogenic fragment in the polarized retinal pigment epithelial cell model of age-related macular degeneration.

Chen Chia-Yi CY Melo Esther E Jakob Peter P Friedlein Arno A Elsässer Brigitta B Goettig Peter P Kueppers Verena V Delobel Frederic F Stucki Corinne C Dunkley Tom T Fauser Sascha S Schilling Oliver O Iacone Roberto R

Matrix biology : journal of the International Society for Matrix Biology 20180320

Age-related macular degeneration (AMD) is the leading cause of irreversible blindness in the elderly population. Variants in the HTRA1-ARMS2 locus have been linked to increased AMD risk. In the present study we investigated the impact of elevated HtrA1 levels on the retina pigment epithelial (RPE) secretome using a polarized culture system. Upregulation of HtrA1 alters the abundance of key proteins involved in angiogenesis and extracellular matrix remodeling. Thrombospondin-1, an angiogenesis mo ...[more]

PMID: 29572155

Similar Datasets

N-terminomic analysis of RPE secretome

Project description:Identification of HtrA1 substrate in the context of RPE secretome

2018-04-09 | PXD007139 | Pride

Murine RPE proteome and phosphoproteome

Project description:One of the major biological functions accomplished by the retinal pigmented epithelium (RPE) is the clearance of shed photoreceptor outer segments (POS) through a multistep process referred to as phagocytosis. Phagocytosis helps maintain the viability of photoreceptors which otherwise could succumb to the high metabolic flux and photo-oxidative stress associated with visual processing. Regulatory mechanisms underlying phagocytosis in the RPE are not fully understood, although dysfunction of this process contributes to the pathogenesis of multiple human retinal degenerative disorders, including age-related macular degeneration (AMD). Here we present an integrated analysis of phagocytosing cultured-RPE cells.

2017-10-27 | PXD006124 | Pride

Polarized RPE Secretome Preserves Photoreceptors in Retinal Dystrophic RCS Rats.

Project description:Retinal degenerative diseases, including age-related macular degeneration (AMD) and retinitis pigmentosa, lack effective therapies. Conventional monotherapeutic approaches fail to target the multiple affected pathways in retinal degeneration. However, the retinal pigment epithelium (RPE) secretes several neurotrophic factors addressing diverse cellular pathways, potentially preserving photoreceptors. This study explored human embryonic stem cell-derived, polarized RPE soluble factors (PRPE-SF) as a combination treatment for retinal degeneration. PRPE-SF promoted retinal progenitor cell survival, reduced oxidative stress in ARPE-19 cells, and demonstrated critical antioxidant and anti-inflammatory effects for preventing retinal degeneration in the Royal College of Surgeons (RCS) rat model. Importantly, PRPE-SF treatment preserved retinal structure and scotopic b-wave amplitudes, suggesting therapeutic potential for delaying retinal degeneration. PRPE-SF is uniquely produced using biomimetic membranes for RPE polarization and maturation, promoting a protective RPE secretome phenotype. Additionally, PRPE-SF is produced without animal serum to avoid immunogenicity in future clinical development. Lastly, PRPE-SF is a combination of neurotrophic factors, potentially ameliorating multiple dysfunctions in retinal degenerations. In conclusion, PRPE-SF offers a promising therapeutic candidate for retinal degenerative diseases, advancing the development of effective therapeutic strategies for these debilitating conditions.

| S-EPMC10340490 | biostudies-literature

Cathepsin B and Cathepsin Z tumor secretome of murine breast cancers

Project description:MS-based proteomics study of the cancer secretome and tumor interstitial fluid of tumor cell/macrophage co-cultures and breast tumors in cells and mice lacking both cathepsin B and cathepsin Z

2021-09-08 | PXD015209 | Pride

RPE Redifferentiation (miRNA)

Project description:Retinal Pigment Epithelial (RPE) cells are located behind the retina and are critical for photoreceptor survival. Loss of RPE is associated with several pathogenic conditions such as Age Related Macular Degeneration and Retinitis Pigmentosa. RPE derived from human embryonic stem cells (hESC) offer a potential source for producing these cells for therapy. Here we report the molecular and cellular characterization of RPE differentiated from hESC. hESC derived RPE are capable of proliferation and lose their epithelial characteristics before becoming confluent and re-differentiating back into their typical pigmented, cobblestoned appearance. During the proliferative phase, they adopt a mesenchymal morphology and express mesenchymal markers. Our results demonstrate that this apparent Epithelial-Mesenchymal Transition is not regulated by the classical EMT transcription factors SNAIL and SLUG. Furthermore, it is possible to regulate RPE de-differentiation and re-differentiation by modulating the Wnt and BMP pathway respectively. These findings further our understanding of the genesis and expansion of RPE which is essential for their therapeutic use.

2013-01-17 | E-MTAB-960 | biostudies-arrayexpress

MicroRNAs of the RPE are essential for RPE differentiation and photoreceptor maturation (mRNA)

Project description:Dysfunction of the retinal pigmented epithelium (RPE) results in degeneration of photoreceptors and vision loss and is correlated with common blinding disorders in humans. Although many protein-coding genes are known to be expressed in RPEs and important for their development and maintenance, virtually nothing is known about the in vivo roles of non-protein coding transcripts in RPEs. The expression patterns of microRNAs (miRNAs) have been analyzed in a variety of ocular tissues, and few were implicated to play role in RPE based on studies in cell lines. Herein, through RPE specific conditional mutagenesis of Dicer1 or DGCR8, the importance of miRNA for RPE differentiation was uncovered. Interestingly, miRNAs were found to be dispensable for maintaining the RPE fate and survival, and yet they are essential for acquisition of important RPE properties such as the expression of genes involved in the visual cycle pathway, pigmentation and cell adhesion. Importantly miRNAs of the RPE were found to be required for maturation of the adjacent photoreceptors, specifically for the morphogenesis of the outer segments. The profiles of miRNA and mRNA altered in the Dicer1 deficient RPE point to a key role of miR-204 in regulation of RPE differentiation program in vivo and uncovers the importance of additional novel RPE miRNAs. The study exposes the combined regulatory activity of miRNAs of the RPE, which is required for RPE differentiation and for the development of the adjacent neuroretina. Effect of Dicer 1 deficiency on RPE miRNA and mRNA.

2015-06-16 | E-GEOD-69881 | biostudies-arrayexpress

MicroRNAs of the RPE are essential for RPE differentiation and photoreceptor maturation (microRNA)

2015-06-16 | E-GEOD-69882 | biostudies-arrayexpress

Expresion data from primary retinal pigment epithelium (RPE) and immortalized RPE

Project description:We used microarrays to detail the global gene expression of primary RPE and immortalized RPE. We investigated highly expression genes in immortalized RPE.

2017-09-07 | GSE80985 | GEO

Human RPE LC-MSMS

Project description:AMD is a complex disorder related to environmental factors and genetic polymorphisms. We explored the interplay between the two factors by comparing the differences between HTRA1/ARMS2 high-risk and low-risk AMD alleles after exposure to smokers’ serum using iTRAQ-based proteomics. Primary human retinal pigment epithelial (hRPE) cells were obtained from donor eyes and screened for ARMS2/HTRA1 AMD risk genotypes, wild type allele as control. A gene ontology annotation analysis and KEGG metabolic pathway analysis were employed to obtain a global functional view of differentially expressed (DE) proteins. Significant DE proteins were screened and validated by real-time PCR, immunofluorescence and western blot. The relationship between HTRA1 and caveolin-1 protein was explored by HTRA1-overexpressing lentivirus-transduced ARPE-19 cells. A total of 400 DE proteins were detected for the high-risk allele after cigarette sera stimulation, which was much higher than the low-risk allele. The caveolin-1 protein was significantly up-regulated when HTRA1-overexpressing cells were exposed to smokers’ serum and the smoking effect was stronger than AMD high-risk allele. There were interplays between AMD high-risk allele and cigarette smoking. Smokers’ sera also accelerated the phagocytosis of hRPE cells. In addition, caveolin-1 may play an important role in the disease process, but its molecular mechanism requires further study.

2022-02-22 | PXD009309 | Pride

RPE-1 and RPE-1 trisomic AHA p-c and steady state data

Project description:We developed a pulse-chase method in where fully SILAC (heavy, medium-heavy and Light) labelled cells were pulsed with Azidohomoalanine (AHA) and then chased for different length of times. In addition, steady state protein levels comparing the RPE- and RPE-1 trisomic cells by standard SILAC labeling was also acquired. These datasets contain data for the human RPE-1 and RPE-1 trisomic cell lines.

2017-03-31 | MSV000080831 | MassIVE