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Published on: May 27, 2016
Exosomal miRNA Profiling in Liquid Biopsy of Vitreous in Proliferative Diabetic Retinopathy
Bin Yan1,2, Jianing Qiu1,2, Yan Yang1,2
1Department of Ophthalmology, the Second Xiangya Hospital, Central South University, Changsha, China.
Purpose:
Proliferative diabetic retinopathy (PDR) is a severe microvascular complication of diabetes mellitus. Although exosomes participate in various pathologic processes, their role in ocular pathologies is unclear. This study profiled exosomal microRNAs (exo-miRNAs) in the vitreous humor (VH) of patients with PDR to elucidate their regulatory roles in PDR pathogenesis.
Methods:
VH samples were obtained from 23 patients with PDR and 17 non-diabetic controls. Exosomes were isolated using ultracentrifugation and characterized via transmission electron microscopy, nanoparticle tracking analysis and Western blot. High-throughput sequencing identified differentially expressed miRNAs (DEMs), followed by target gene prediction and bioinformatic analyses. The top DEMs were validated using quantitative real-time polymerase chain reaction (qRT-PCR).
Results:
Exosomes were isolated and characterized from VH samples. Sequencing analysis identified 843 unique miRNAs, with 60 showing significant differential expression between PDR and control groups (28 upregulated and 32 downregulated). The miR-451a and miR-486-5p were most upregulated; miR-204-5p and miR-211-5p were most downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed DEM target genes enriched in endocrine, metabolic, signaling transduction, and intercellular adhesion pathways, notably cGMP-PKG, mTOR, and cAMP pathways. Network analysis identified miR-486-5p and miR-451a as key hubs. Validation by qRT-PCR confirmed the differential expression in the sequencing analysis.
Conclusions:
This study provides the comprehensive profile of vitreous exo-miRNAs in patients with PDR, identifying key DEMs and their potential regulatory networks. These findings indicate that vitreous exo-miRNAs, specifically miR-204-5p and miR-486-5p, represent promising diagnostic biomarkers and potential therapeutic targets for PDR.
Insights
Researchers identified specific exosomal microRNAs (exo-miRNAs) in vitreous humor linked to proliferative diabetic retinopathy (PDR). These exo-miRNAs, including miR-204-5p and miR-486-5p, show potential as diagnostic biomarkers and therapeutic targets for PDR.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Proliferative diabetic retinopathy (PDR) is a severe complication of diabetes mellitus.
- The role of exosomes and exosomal microRNAs (exo-miRNAs) in ocular pathologies like PDR is not well understood.
- Understanding these roles is crucial for developing new diagnostic and therapeutic strategies.
Purpose of the Study:
- To profile exo-miRNAs in the vitreous humor (VH) of patients with PDR.
- To elucidate the potential regulatory roles of these exo-miRNAs in PDR pathogenesis.
- To identify potential diagnostic biomarkers and therapeutic targets for PDR.
Main Methods:
- Vitreous humor samples were collected from patients with PDR and non-diabetic controls.
- Exosomes were isolated and characterized using ultracentrifugation, transmission electron microscopy, nanoparticle tracking analysis, and Western blot.
- High-throughput sequencing identified differentially expressed miRNAs (DEMs), followed by bioinformatic analyses and qRT-PCR validation.
Main Results:
- Sequencing identified 843 unique miRNAs, with 60 significantly differentially expressed between PDR and control groups.
- Key upregulated exo-miRNAs included miR-451a and miR-486-5p; downregulated included miR-204-5p and miR-211-5p.
- DEM target genes were enriched in metabolic and signaling pathways, with miR-486-5p and miR-451a identified as key regulatory hubs.
Conclusions:
- This study provides a comprehensive profile of vitreous exo-miRNAs in PDR patients.
- Specific exo-miRNAs, notably miR-204-5p and miR-486-5p, are implicated in PDR pathogenesis.
- These exo-miRNAs represent promising diagnostic biomarkers and potential therapeutic targets for PDR.
