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Normalization strategies differently affect circulating miRNA profile associated with the training status.

Martina Faraldi1, Marta Gomarasca2, Veronica Sansoni2

  • 1Laboratory of Experimental Biochemistry & Molecular Biology, IRCCS Istituto Ortopedico Galeazzi, Milano, Italy. martina.faraldi@grupposandonato.it.

Scientific Reports
|February 9, 2019
PubMed
Summary
This summary is machine-generated.

Standardizing microRNA (miRNA) analysis is crucial for biomarker development. This study identified hsa-miR-320d as a reliable reference miRNA for analyzing circulating miRNA profiles in athletes versus sedentary individuals.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Exercise Physiology

Background:

  • MicroRNAs (miRNAs) are key regulators of adaptive responses.
  • Circulating miRNAs show potential as biomarkers for physiological status.
  • Standardized analytical procedures are lacking for miRNA biomarker use.

Purpose of the Study:

  • To compare normalization methods for RT-qPCR analysis of circulating miRNAs.
  • To identify a reliable normalization strategy for sedentary and highly-trained subjects.
  • To investigate circulating miRNA profiles associated with physical activity.

Main Methods:

  • Plasma RNA extraction followed by RT-qPCR assay of 179 miRNAs.
  • Application of various normalization strategies using endogenous miRNAs and exogenous oligonucleotides.
  • Comparison of data outputs from different normalization approaches.

Main Results:

  • hsa-miR-320d demonstrated the lowest technical variability, making it a suitable reference miRNA.
  • Significant differences in circulating miRNA profiles were observed between sedentary and highly-trained individuals.
  • Physically active status is associated with specific skeletal muscle- and bone-related circulating miRNAs.

Conclusions:

  • Accurate normalization is essential to avoid misinterpretation of circulating miRNA expression data.
  • hsa-miR-320d is a reliable reference for analyzing circulating miRNAs in exercise studies.
  • Circulating miRNA profiles reflect epigenetic modifications related to physical activity and tissue homeostasis.