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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
Published on: April 14, 2015
Micro-array profiling exhibits remarkable intra-individual stability of human platelet micro-RNA
C Stratz1, T G Nührenberg, H Binder
1Bernd L. Fiebich, Department of Psychiatry and Psychotherapy, University of Freiburg Medical School, Hauptstr. 5, D-79014 Freiburg, Germany.
Abstract:
Platelets play an important role in haemostasis and thrombus formation. Latest research identified platelets harbouring so called microRNAs (miRNA). MiRNAs are short single-stranded RNAs modulating gene expression by targeting mRNAs. Limited data exist on inter-individual variability of platelet miRNA profile while no data are available on intra-individual variability. We assessed platelet miRNA profile in five volunteers at five time points over a time course of 10 days; 24 hours prior to the last blood sampling, subjects took 500 mg acetylsalicylic acid (ASA). Platelet miRNA was isolated from leucocyte-depleted platelet-rich plasma, and miRNA array-analysis was performed. Temporal patterns and ASA effect were explored by a linear mixed effects model for each miRNA. For the 20 most abundantly expressed platelet miRNAs, target gene search was performed and an annotation network was created. MiRNA expression profiling of 1,281 human miRNAs revealed relevant expression of 221 miRNAs consistently expressed in all samples at all time points. Correlation of platelet miRNA ranks was highly significant to other studies. Global distribution of miRNA expression was relatively similar in all subjects. No miRNA exhibited a significant effect of time at level 0.05. After 24 hours, no significant effect of ASA was found. Concerning functional implications of the 20 most abundantly expressed miRNAs, we found six functional themes. In conclusion, platelet miRNA profile is remarkably stable over the time period studied. Single-point analysis of platelet miRNA profile is reasonable when inter-individual differences are studied. The functional annotation network points toward extra-platelet effects of platelet miRNAs.
Insights
Platelet microRNA (miRNA) profiles are stable over time and across individuals. This stability supports single-point analysis for studying inter-individual differences in platelet miRNA expression.
Area of Science:
- Molecular Biology
- Hematology
- Genomics
Background:
- Platelets are crucial for hemostasis and thrombosis.
- Platelets contain microRNAs (miRNAs), which regulate gene expression.
- Limited data exist on platelet miRNA variability, especially intra-individual.
Purpose of the Study:
- To assess the intra-individual variability of platelet miRNA profiles over 10 days.
- To investigate the effect of acetylsalicylic acid (ASA) on platelet miRNA expression.
- To explore the functional implications of abundant platelet miRNAs.
Main Methods:
- Platelet miRNA was isolated from leucocyte-depleted platelet-rich plasma in five volunteers over 10 days.
- miRNA array analysis was performed on 1,281 human miRNAs.
- A linear mixed effects model analyzed temporal patterns and ASA effects; target gene networks were created for abundant miRNAs.
Main Results:
- 221 miRNAs were consistently expressed across all samples and time points.
- No significant effect of time or 24-hour acetylsalicylic acid (ASA) administration on miRNA expression was detected.
- Functional annotation revealed six themes for the 20 most abundant platelet miRNAs, suggesting extra-platelet roles.
Conclusions:
- Platelet miRNA profiles demonstrate remarkable stability over a 10-day period.
- Single-point analysis is a valid approach for studying inter-individual differences in platelet miRNA.
- Platelet miRNAs may exert functions beyond the platelet itself.
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