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Stability of Circulating Exosomal miRNAs in Healthy Subjects

David Sanz-Rubio1, Inmaculada Martin-Burriel2, Ana Gil3

  • 1Translational Respiratory Research Unit, IISAragon & CIBERES, Hospital Universitario Miguel Servet, Zaragoza, Spain. davidsanzrubio91@gmail.com.

Scientific Reports
|July 10, 2018
PubMed

Insights

This study compared exosome isolation methods and assessed plasma exosomal microRNA (miRNA) stability. Freezing plasma before isolation yielded higher miRNA levels and lower variability, establishing exosomal miRNAs as reliable biomarkers.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Exosomes are crucial for intercellular communication and contain microRNAs (miRNAs).
  • Circulating exosomal miRNAs are promising biomarkers, but their isolation and stability require further investigation.
  • Variability in exosomal miRNA cargo and isolation methods impacts biomarker reliability.

Purpose of the Study:

  • To compare two exosome isolation methods for plasma.
  • To assess the stability of eleven plasma exosomal miRNAs over time.
  • To evaluate the effect of pre-isolation plasma freezing versus post-isolation exosome freezing on miRNA stability.

Main Methods:

  • Exosome isolation from plasma using two different kits.
  • Quantification of eleven exosomal miRNAs via quantitative PCR.
  • Analysis of miRNA stability across four consecutive weekly blood draws from 7 healthy subjects.
  • Evaluation of inter-individual and intra-individual miRNA variability.

Main Results:

  • One isolation kit demonstrated superior amplification signals.
  • Isolating exosomes from previously frozen plasma resulted in higher miRNA levels.
  • Pre-isolation plasma freezing led to lower intra-subject and inter-subject miRNA variability.
  • Selected miRNAs exhibited stability across consecutive weekly extractions.

Conclusions:

  • Exosome isolation from frozen plasma enhances miRNA yield and stability.
  • This study demonstrates the temporal stability of plasma exosomal miRNAs.
  • These findings are critical for establishing exosomal miRNAs as robust diagnostic biomarkers.

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