Nano-seq analysis reveals different functional tendency between exosomes and microvesicles derived from hUMSC.

Area of Science:

• Stem Cell Biology
• Extracellular Vesicles
• RNA Biology
• Nanopore Sequencing

Background:

• Extracellular vesicles (EVs) from human umbilical cord mesenchymal stem cells (hUMSCs) are key for cell-free therapy.
• Understanding EV RNA composition is crucial for safe and precise applications.
• Current RNA analysis methods (NGS) limit comprehensive profiling of long transcripts.

Purpose of the Study:

• To profile the long transcriptomic landscape of hUMSC-EVs using nanopore sequencing.
• To compare the transcriptomic profiles of different hUMSC-EV subtypes (exosomes and microvesicles).
• To identify distinct RNA signatures and potential functions of EV subtypes.

Main Methods:

• Isolation and identification of exosome and microvesicle subtypes from hUMSCs via density gradient centrifugation.
• Systematic comparison of long transcriptomic profiles using nanopore sequencing.
• Bioinformatic analysis of transcriptomic data.

Main Results:

• Nanopore sequencing identified abundant transcript variants, with 69.34% being fragmented.
• Exosomes showed a higher proportion of intact/near-complete transcripts compared to microvesicles.
• Distinct gene enrichment pathways were observed: neurodegenerative diseases in microvesicles and neutrophil extracellular trap formation in exosomes.

Conclusions:

• This study provides novel insights into the distinct transcriptomic profiles of hUMSC-EV subtypes.
• Findings suggest different RNA sorting mechanisms and functional roles for exosomes and microvesicles.
• Different EV subtypes from the same source exhibit unique physiological functions, indicating diverse clinical application prospects.