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Published on: January 31, 2018
MicroRNA 320a and Membrane Antigens as Tools to Evaluate the Pathophysiology of Platelets Stored in Blood Banks
Priscilla Cristina Moura Vieira1,2, Jersey Heitor da Silva Maués3, Letícia Martins Lamarão4
1Human Cytogenetics Laboratory, Biological Science Institute, Federal University of Pará, Belém 66075-110, PA, Brazil.
Abstract:
Our research group, through the analysis of miRNomes in platelet concentrates (PCs) stored in blood banks, identified and validated the miR-127 and miR-320a miRNAs as biomarkers of platelet storage lesions (PSLs) in PCs. In order to validate the miRNAs 127 and 320a methodologically, as PSL biomarkers in a large number of PC bags, we also evaluated important immunological markers involved in the platelet activation/aggregation process-the CD62P receptor (P-selectin), the surface glycoproteins (GP) IIb/IIIa, and the purinergic P2Y12 receptor-via flow cytometry. The miRNAs miR-127 and miR-320a were quantified by real-time quantitative PCR (RT-qPCR). To carry out this study, 500 collection tubes were used at the upper edge of the PC bags containing platelets. Each tube was divided into seven equal parts (totaling 3500 samples) for platelet analysis from 7 different storage days, where the 1st day represents the high-quality control, and the 7th day corresponds to the low-quality control of the platelets. After analyzing all parameters during storage days, it was concluded that the relative quantification of miR-320a below 0.50 and the CD62P receptor below 27.92% are reliable indicators of the absence of storage lesions in blood banks. We believe that the values found in the expression of the CD62P receptor legitimize the use of the miR-320a and miR-127 miRNAs to build a kit capable of accurately measuring whether the stored platelets are suitable for transfusion.
Insights
Researchers identified miR-320a and miR-127 as reliable biomarkers for detecting platelet storage lesions (PSLs) in blood banks. Low levels of miR-320a and CD62P receptor indicate safe platelets for transfusion.
Area of Science:
- Biochemistry
- Molecular Biology
- Hematology
Background:
- Platelet concentrates (PCs) are crucial for transfusions, but storage can lead to platelet storage lesions (PSLs).
- Accurate biomarkers are needed to assess PC quality and ensure transfusion safety.
- Current methods for evaluating PC quality are limited.
Purpose of the Study:
- To identify and validate microRNAs (miRNAs) as biomarkers for PSLs in stored PCs.
- To correlate miRNA expression with immunological markers of platelet activation.
- To establish reliable indicators for assessing the suitability of PCs for transfusion.
Main Methods:
- Analysis of miRNomes in PCs using real-time quantitative PCR (RT-qPCR) to quantify miR-127 and miR-320a.
- Flow cytometry to evaluate immunological markers: CD62P receptor, GP IIb/IIIa, and P2Y12 receptor.
- Study involved 3500 samples from 500 collection tubes across 7 storage days.
Main Results:
- miR-127 and miR-320a were identified as potential biomarkers for PSLs.
- Relative quantification of miR-320a below 0.50 is a reliable indicator of absent PSLs.
- CD62P receptor expression below 27.92% also indicates the absence of storage lesions.
Conclusions:
- miR-320a and miR-127 are validated biomarkers for assessing platelet quality in blood banks.
- The combination of miR-320a levels and CD62P receptor expression can accurately determine PC suitability for transfusion.
- These findings support the development of a diagnostic kit for stored platelet quality assessment.
