The proteome of normal human retrobulbar optic nerve and sclera

Zhang, Pingbo; Karani, Rabia; Turner, Randi; Dufresne, Craig; Ferri, Sara; Eyk, Jennifer E. Van; Semba, Richard D.

doi:10.1002/pmic.201600229

Cited by 16 publications

(14 citation statements)

References 20 publications

(25 reference statements)

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“…They were subjected to blast analysis to find conserved retinal-specific proteins. [8][9][10]14 Further, ID match analysis was performed between dataset 1 (609 proteins) and dataset 2 (672 proteins) to identify dataset 3 (338 proteins) consisting of conserved corneal specific proteins. Further, the corresponding sequences were batch retrieved from Uniprot database.…”

Section: Peptide Designing Strategymentioning

confidence: 99%

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Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Amit¹,

Muralikumar²,

Janaki³

et al. 2019

IJN

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BackgroundFungal keratitis is a major cause of corneal blindness accounting for more than one-third of microbiologically proven cases. The management of fungal keratitis is through topical or systemic antifungal medications alone or in combination with surgical treatment. Topical medications such as natamycin and voriconazole pose major challenges due to poor penetration across the corneal epithelium. To address the issue various carrier molecules like nanoparticles, lipid vesicles, and cell penetrating peptides were explored. But the major drawback such as non-specificity and lack of bioavailability remains.PurposeIn this study, we have attempted to design corneal specific cell penetrating peptide using subtractive proteomic approach from the published literature and tried to improve its bioavailability through gelatin hydrogel delivery system.Material and MethodsUsing subtractive proteomic approach two peptides VRF005 and VRF007 were identified on the basis of solubility, cell permeability and amphipathicity. The peptides were modeled for three-dimensional structure and simulated for membrane penetration. The peptides were characterized using circular dichroism spectroscopy, dynamic light scattering and native polyacrylamide gel electrophoresis. Further uptake studies were performed on primary corneal epithelial cells and the stability was analyzed in corneal epithelial tissue lysates. Insilico prediction of peptides showed it to have antifungal activity which was further validated using colony forming assay and time killing kinetics. The duration of antifungal activity of peptide was improved using gelatin hydrogel through sustained delivery.ResultsVRF005 and VRF007 showed α-helical structure and was within the allowed region of Ramachandran plot. The simulation study showed their membrane penetration. The peptide uptake was found to be specific to corneal epithelial cells and also showed intracellular localization in Candida albicans and Fusarium solani. Peptides were found to be stable up to 2 hours when incubated with corneal epithelial tissue lysate. Dynamic light scattering, and native polyacrylamide gel electrophoresis revealed aggregation of peptides. VRF007 showed antifungal activity up to 24 hour whereas VRF005 showed activity up to 4 hours. Hence gelatin hydrogel-based delivery system was used to improve the activity. Actin staining of corneal epithelial cells showed that the cells were attached on gelatin hydrogel.ConclusionWe have designed corneal specific cell penetrating peptides using subtractive proteomic approach. Bioavailability and delivery of peptide was enhanced using gelatin hydrogel system.

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Section: Peptide Designing Strategymentioning

confidence: 99%

“…Semba et al 7 published a series of articles on human eye proteome [8][9][10] listing out the set of abundant proteins in different parts of the eye (choroid, retina, lens, iris, ciliary body, etc). Dyrlund et al 11 identified and quantified the list of proteins in the layer of the cornea.…”

Section: Introductionmentioning

confidence: 99%

Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Amit¹,

Muralikumar²,

Janaki³

et al. 2019

IJN

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show abstract

“…Nonetheless, this observation needs to be substantiated by further experiments. Further, we analyzed the schisis fluid-specific proteins with the proteome data sets of various ocular tissues such as the retina, ciliary body, iris, retinal pigment epithelium, choroid, sclera and optic nerve [23–25]. On comparison, we found that 257 (33%) schisis fluid proteins were not detected in any of the ocular tissues including the vitreous fluid.…”

Section: Discussionmentioning

confidence: 99%

Proteomic profiling of human intraschisis cavity fluid

et al. 2017

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BackgroundX-linked retinoschisis (XLRS) is a vitreoretinal degenerative disorder causing vision deterioration, due to structural defects in retina. The hallmark of this disease includes radial streaks arising from the fovea and splitting of inner retinal layers (schisis). Although these retinal changes are attributed to mutations in the retinoschisin gene, schisis is also observed in patients who do not carry mutations. In addition, the origin of intraschisis fluid, the triggering point of schisis formation and its progression are largely unknown still. So far, there is no report on the complete proteomic analysis of this fluid. Schisis fluid proteome could reflect biochemical changes in the disease condition, helping in better understanding and management of retinoschisis. Therefore it was of interest to investigate the intraschisis fluid proteome using high-resolution mass spectrometry.MethodsTwo male XLRS patients (aged 4 and 40 years) underwent clinical and genetic evaluation followed by surgical extraction of intraschisis fluids. The two fluid samples were resolved on a SDS-PAGE and the processed peptides were analyzed by Q-Exactive plus hybrid quadrupole-Orbitrap mass spectrometry. Functional annotation of the identified proteins was performed using Ingenuity pathway analysis software.ResultsMass spectrometry analysis detected 770 nonredundant proteins in the intraschisis fluid. Retinol dehydrogenase 14 was found to be abundant in the schisis fluid. Gene ontology based analysis indicated that 19% of the intraschisis fluid proteins were localized to the extracellular matrix and 15% of the proteins were involved in signal transduction. Functional annotation identified three primary canonical pathways to be associated with the schisis fluid proteome viz., LXR/RXR activation, complement system and acute phase response signalling, which are involved in immune and inflammatory responses. Collectively, our results show that intraschisis fluid comprises specific inflammatory proteins which highly reflect the disease environment.ConclusionBased on our study, it is suggested that inflammation might play a key role in the pathogenesis of XLRS. To our knowledge, this is the first report describing the complete proteome of intraschisis fluid, which could serve as a template for future research and facilitate the development of therapeutic modalities for XLRS.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-017-9148-y) contains supplementary material, which is available to authorized users.

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“…Earlier reports also used mass spectrometry‐based proteomics to profile the ocular surface. In particular, the Human Eye Proteome Project aims to characterize all sub‐proteomes of the healthy eye and profile the tear proteome . Additional studies have also profiled ocular protein level differences following chemical exposure and during macular degeneration .…”

Section: Introductionmentioning

confidence: 99%

Label‐Free Quantitative Proteomics Distinguishes General and Site‐Specific Host Responses to Pseudomonas aeruginosa Infection at the Ocular Surface

2020

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Mass spectrometry-based proteomics enables the unbiased and sensitive profiling of cellular proteomes and extracellular environments. Recent technological and bioinformatic advances permit identifying dual biological systems in a single experiment, supporting investigation of infection from both the host and pathogen perspectives. At the ocular surface, Pseudomonas aeruginosa is commonly associated with biofilm formation and inflammation of the ocular tissues, causing damage to the eye. The interaction between P. aeruginosa and the immune system at the site of infection describes limitations in clearance of infection and enhanced pathogenesis. Here, the extracellular environment (eye wash) of murine ocular surfaces infected with a clinical isolate of P. aeruginosa is profiled and neutrophil marker proteins are detected, indicating neutrophil recruitment to the site of infection. The first potential diagnostic markers of P. aeruginosa-associated keratitis are also identified. In addition, the deepest murine corneal proteome to date is defined and proteins, categories, and networks critical to the host response are detected. Moreover, the first identification of bacterial proteins attached to the ocular surface is reported. The findings are validated through in silico comparisons and enzymatic profiling. Overall, the work provides comprehensive profiling of the host-pathogen interface and uncovers differences between general and site-specific host responses to infection.

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The proteome of normal human retrobulbar optic nerve and sclera

Cited by 16 publications

References 20 publications

Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Designing and enhancing the antifungal activity of corneal specific cell penetrating peptide using gelatin hydrogel delivery system

Proteomic profiling of human intraschisis cavity fluid

Label‐Free Quantitative Proteomics Distinguishes General and Site‐Specific Host Responses to Pseudomonas aeruginosa Infection at the Ocular Surface

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