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Proteomic analysis of vitreal exosomes in patients with proliferative diabetic retinopathy

Eye volume 37, pages 2061–2068 (2023)Cite this article

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Abstract

Purpose

To determine the proteomic profiles of exosomes derived from vitreous humour (VH) obtained from proliferative diabetic retinopathy (PDR) patients and non-diabetic controls with idiopathic macular hole/epiretinal membrane.

Methods

Vitreal exosomes were isolated using differential ultracentrifugation, followed by characterisation performed using different techniques. A label-free proteomic analysis was conducted to determine the protein profiles of the exosomes. A parallel reaction monitoring (PRM) analysis was performed to verify the identified proteins and associated functional annotations were derived by gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Receiver operating characteristic (ROC) analysis was utilised to evaluate the diagnostic value of target proteins in distinguishing PDR from controls.

Results

Exosomes were successfully isolated from VH, and were well characterised by various techniques. The results of proteomic analysis showed that a total of 758 proteins were identified and 10 proteins were screened as differentially expressed proteins, significantly changed in the PDR group containing 4 elevated proteins and 6 reduced proteins. GO analysis indicated that these differential proteins were mainly involved in many metabolic pathways, including nicotinamide adenine dinucleotide metabolism, adenosine diphosphate metabolic process and glycolytic process. The KEGG analysis enriched the top five pathways including glycolysis/gluconeogenesis, fructose and mannose metabolism, biosynthesis of amino acids, hypoxia-inducible factor 1 signalling pathway and carbon metabolism. The differential proteins, namely, lactate dehydrogenase A, ficolin 3, apolipoprotein B and apolipoprotein M, were further verified by PRM and showed a consistent trend with label-free proteomic analysis. The ROC analysis identified these proteins as promising biomarkers for PDR diagnosis.

Conclusions

Vitreal exosomes from patients with PDR contained few proteins unique to PDR; thus, exosomal proteins have great potential as disease biomarkers and therapeutic targets for PDR.

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Fig. 1: An overview of the experimental design.
Fig. 2: Characterisation of isolated vitreous exosomes across groups.
Fig. 3: Hierarchical cluster and volcano plots of the differentially expressed proteins.
Fig. 4: Parallel reaction monitoring (PRM) analysis of the differentially expressed exosome proteins.

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Data availability

The data that support the findings of this study are not publicly available because they contain information that could compromise the privacy of patients but are available from the corresponding author upon reasonable request.

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Funding

This study was supported by the Jinan Clinical Medical Science and Technology Innovation Plan (202019113), Key Research and Development project of Shandong Province (2017GSF218033) and the National Natural Science Foundation of China (81700831).

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Author notes

  1. These authors contributed equally: Jiawei Wang, Zhenzhen Wang.

Authors and Affiliations

  1. Department of Ophthalmology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China

    Jiawei Wang, Zhenzhen Wang, Ying Zhang & Jianqiao Li

  2. Liaocheng Eye Hospital, Liaocheng, China

    Zhenzhen Wang

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  1. Jiawei Wang
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Contributions

All authors conceived of and designed the experimental protocol. JWW, ZZW and YZ collected the data. JWW and ZZW were involved in the analysis and interpretation of the data. JWW wrote the first draft of the manuscript. JQL reviewed and revised the manuscript and produced the final version. All authors read and approved the final manuscript.

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Correspondence to Jianqiao Li.

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Wang, J., Wang, Z., Zhang, Y. et al. Proteomic analysis of vitreal exosomes in patients with proliferative diabetic retinopathy. Eye 37, 2061–2068 (2023). https://doi.org/10.1038/s41433-022-02286-x

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