MiRNAs:Biomarkers of Storage Lesions of RBC.
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus Rattus norvegicus Homo sapiens
PROVIDER: GSE86905 | GEO | 2016/09/15
SECONDARY ACCESSION(S): PRJNA342939
REPOSITORIES: GEO
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Project description:Mature human red blood cells (RBCs) are terminally differentiated anuclear cells. While initially thought to lack any nucleic acids, human RBCs are found to contain abundant and diverse species of RNA transcripts with functional relevance. Given the absence of novel transcription, RBCs may provide an interesting cellular context to study RNA metabolism over time. One clinically relevant context is the ex vivo storage of RBCs in blood banks for use in blood transfusion. Some studies have indicated that the transfusion of “old” or aged stored RBCs may be associated with adverse outcomes due to various storage changes termed “storage lesions”. However, other studies do not support these effects, and much remains unknown about the relevant changes associated with RBC storage. Here, we employed the NanoString nCounter assay for global miRNA profiling to comprehensively define the miRNA turnover during ex vivo RBC storage. This profiling demonstrates that the abundance of most RBC miRNAs did not change significantly during the 42 days of refrigerated storage, indicating extremely long decay half-lives. Unexpectedly, miR-720, a cleavage product of tRNAThr, increased dramatically in the first two weeks and persisted during storage. Furthermore, we present evidence for a role of angiogenin in tRNA cleavage to generate miR-720 during RBC storage. The dramatic increase in miR-720 may serve as a new characteristic for storage lesion and may be used to monitor transfused RBCs in clinical patients and athletes performing blood doping.
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Project description:Investigation of miRNA expression level changes of human platelets stored for 2, 5 and 8 days，and to explore the mechanism of storage lesions of human platelets.
2018-09-20 | GSE120187 | GEO
Project description:Phosphorylation of proteins is involved in cell functions, of which cytoskeleton organization. In particular, the phosphotyrosine (pY) has been reported to play a role in red blood cell (RBC) functions. During the storage of RBC in the context of transfusion medicine, cells suffer from storage lesions that affect energy metabolism as well as cell morphology. In the present research work, RBCs were treated (in absence of calcium) with a protein tyrosine phosphatase inhibitor (orthovanadate, OV) to trigger phosphorylation allowing to investigate this effect on RBCs during the storage. Erythrocytes were studied by phosphoproteomics, Western blot analyses and cell morphology by digital holographic microscopy in presence of kinase inhibitors. The OV treatment triggered phosphorylation events, that disappeared during the storage. As a result, 609 phosphoproteins containing 5’298 phosphosites were detected in the membrane extract, of which 43 pY were up- or down-regulated. Following these phosphorylation processes, shape of RBCs shifted from discocytes to spherocytes, and the addition of kinase inhibitors (KIs) inhibited this transition by counteracting the OV treatment. In addition, the different KIs used highlighted potentially two mechanisms involving membrane proteins and playing a role in RBC shape. The capacity of RBCs to maintain their function is central in transfusion medicine and the presented results contribute to a better understanding of RBC biology.
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Project description:We conducted an animal experiment and used total RNA sequencing (RNA-Seq) to identify novel RNA markers to detect autologous blood transfusion (ABT) doping within red blood cells (RBCs) as a pilot study before human trials. The total RNA-Seq and bioinformatics were performed using rat RBCs that were stored 10- and 20-day periods with citrate–phosphate–dextrose solution with adenine (CPDA) at 4°C In the results, we identified three significant genes whose expression levels were drastically decreased according to the storage period. Our novel insights may allow future studies to develop a test method for ABT doping.
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Project description:In blood banks, platelets are stored until 7 days after a pathogen reduction technology (PRT) treatment, Mirasol® (vitamin B2 plus UVB light) in the present case. The storage time under these conditions may have an impact on platelets and their releasate leading to potential adverse reactions following transfusion to patients. The aim of this study was to analyze the proteome of extracellular vesicles generated by platelets at different storage days (2 and 7) to gain deeper information on the platelet concentrates state at those moments. EVs were isolated by a centrifugation-based approach and characterized by dynamic light scattering and transmission electron microscopy. Proteomic analysis was by LC-MS/MS and quantification by SWATH. In this way, 151 proteins were found up-regulated at day 7 of storage. This group includes CCL5 and Platelet factor 4, chemokines with power to attract neutrophils and monocytes, which could generate transfusion adverse reactions. In addition, other glycoproteins and platelet activation markers were also found elevated at day 7. Proteins related to glycolysis and lactate production were found altered with high fold changes, showing a deregulation of platelet metabolism at day 7. The obtained results provide novel information about possible effects of platelet-derived EVs on transfusion adverse reactions.
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