Comprehensive Analysis of Serum Small Extracellular Vesicles-Derived Coding and Non-Coding RNAs from Retinoblastoma...

Area of Science:

• Biochemistry
• Molecular Biology
• Genomics

Background:

• Small extracellular vesicles (sEVs) are implicated in intercellular communication.
• Dysregulation of sEVs is observed in various diseases, including Retinoblastoma (RB).
• Understanding the RNA cargo within RB-derived sEVs is crucial for diagnostic and therapeutic insights.

Purpose of the Study:

• To comprehensively characterize serum-derived sEVs from RB patients.
• To identify differentially expressed coding and non-coding RNAs within these sEVs.
• To elucidate the regulatory networks and functional roles of these RNAs in RB.

Main Methods:

• Nanoparticle tracking analysis, transmission electron microscopy, and immunoblotting for sEV characterization.
• RNA sequencing and quantitative real-time PCR for differential RNA expression analysis.
• Bioinformatics for functional analysis and construction of regulatory networks (miRNA-mRNA, lncRNA-miRNA-mRNA, protein-protein interaction).

Main Results:

• Characterized sEVs: round-shaped (<150 nm), high concentration (5.3 × 10^11 particles/mL), expressing exosome markers (CD9, CD81, TSG101).
• Identified significant differential expression in 6514 mRNAs, 123 miRNAs, and 3634 lncRNAs.
• Revealed regulatory networks where ncRNAs (e.g., MALAT1, AFAP1-AS1) target cell cycle genes (e.g., CDKNI1A, c-MYC) and identified eye development-related mRNA functions.

Conclusions:

• Provides a detailed molecular profile of RNAs within RB patient serum sEVs.
• Highlights the regulatory roles of specific ncRNAs in cell cycle progression and eye development pathways.
• Establishes a foundation for understanding RB pathogenesis through sEV-mediated RNA communication.