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Changes in MicroRNA Expression Level of Circulating Platelets Contribute to Platelet Defect After Cardiopulmonary
Nobuhiro Mukai1, Yoshinobu Nakayama1, Sachiyo Ishi1
1Department of Anesthesiology and Critical Care, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Objectives:
Platelet defect mechanisms after cardiopulmonary bypass remain unclear. Our hypothesis microRNA expressions in circulating platelets significantly change between pre and post cardiopulmonary bypass, and consequent messenger RNA and protein expression level alterations cause postcardiopulmonary bypass platelet defect.
Design:
Single-center prospective observational study.
Setting:
Operating room of Kyoto Prefectural University of Medicine.
Patients:
Twenty-five adult patients scheduled for elective cardiac surgeries under cardiopulmonary bypass.
Interventions:
None.
Measurements And Main Results:
In the initial phase, changes in microRNA expression between pre and post cardiopulmonary bypass underwent next generation sequencing analysis (10 patients). Based on the results, we focused on changes in mir-10b and mir-96, which regulate glycoprotein 1b and vesicle-associated membrane protein 8, respectively, and followed them until messenger RNA and protein syntheses (15 patients) using quantitative polymerase chain reaction and Western blotting. Seven microRNAs including mir-10b and mir-96 exhibited significant differences in the initial phase. In the subsequent phase, mir-10b-5p and mir-96-5p overexpressions were confirmed, and glycoprotein 1b and vesicle-associated membrane protein 8 messenger RNA levels were significantly decreased after cardiopulmonary bypass: fold differences (95% CI): mir-10b-5p: 1.35 (1.05-2.85), p value equals to 0.01; mir-96-5p: 1.59 (1.06-2.13), p value equals to 0.03; glycoprotein 1b messenger RNA: 0.46 (0.32-0.60), p value of less than 0.001; and vesicle-associated membrane protein messenger RNA: 0.70 (0.56-0.84), p value of less than 0.001. Glycoprotein 1b and vesicle-associated membrane protein 8 were also significantly decreased after cardiopulmonary bypass: glycoprotein 1b: 82.6% (71.3-93.8%), p value equals to 0.005; vesicle-associated membrane protein 8: 79.0% (70.7-82.3%), p value of less than 0.001.
Conclusions:
Expressions of several microRNAs in circulating platelets significantly changed between pre and post cardiopulmonary bypass. Overexpressions of mir-10b and mir-96 decreased glycoprotein 1b and vesicle-associated membrane protein 8 messenger RNA as well as protein, possibly causing platelet defect after cardiopulmonary bypass.
Insights
Platelet microRNA expressions change after cardiopulmonary bypass, leading to decreased glycoprotein 1b and vesicle-associated membrane protein 8. These alterations in microRNA, messenger RNA, and protein levels likely cause post-cardiopulmonary bypass platelet dysfunction.
Area of Science:
- Cardiovascular Surgery
- Molecular Biology
- Hematology
Background:
- Platelet dysfunction after cardiopulmonary bypass (CPB) is a significant clinical issue with unclear underlying mechanisms.
- MicroRNAs (miRNAs) are key regulators of gene expression and may play a role in CPB-induced platelet defects.
Purpose of the Study:
- To investigate changes in circulating platelet miRNA expression before and after CPB.
- To determine if altered miRNA expression affects the levels of target messenger RNAs (mRNAs) and proteins involved in platelet function.
Main Methods:
- Prospective observational study involving 25 adult patients undergoing elective cardiac surgery with CPB.
- Next-generation sequencing was used to analyze miRNA expression changes in platelets pre- and post-CPB in 10 patients.
- Quantitative PCR and Western blotting were employed to validate changes in specific miRNAs (miR-10b, miR-96) and their target genes (glycoprotein 1b, vesicle-associated membrane protein 8) in 15 patients.
Main Results:
- Seven miRNAs, including miR-10b and miR-96, showed significant expression changes post-CPB.
- Overexpression of miR-10b-5p and miR-96-5p was confirmed post-CPB.
- Significant decreases in glycoprotein 1b and vesicle-associated membrane protein 8 mRNA and protein levels were observed after CPB.
Conclusions:
- Circulating platelet miRNA expression is significantly altered following cardiopulmonary bypass.
- Overexpression of specific miRNAs (miR-10b, miR-96) correlates with reduced levels of key platelet proteins (glycoprotein 1b, vesicle-associated membrane protein 8).
- These molecular changes provide a potential mechanism for the platelet defects observed after cardiopulmonary bypass.
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