MicroRNA Profiling of Exosomes Derived from Red Blood Cell Units: Implications in Transfusion-Related Immunomodulation.

Haobo Huang,Danhui Fu,Liping Fan,Jinfeng Zhu,Shijin Wei,Qiuyan Lin,Biyu Wei

doi:10.1155/2019/2045915

Haobo Huang, Danhui Fu + Show 5 more

Open Access

https://doi.org/10.1155/2019/2045915

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Abstract

Purpose To elucidate the microRNAs existent in exosomes derived from stored red blood cell (RBC) unit and their potential function. Materials and Methods Exosomes were isolated from the supernatant derived from stored RBC units by sequential centrifugation. Isolated exosomes were characterized by TEM (transmission electron microscopy), western blotting, and DLS (dynamic light scattering). MicroRNA (miRNA) microarray was performed to detect the expression of miRNAs in 3 exosome samples. Results revealed miRNAs that were simultaneously expressed in the 3 exosome samples and were previously reported to exist in mature RBCs. Functions and potential pathways of some detected miRNAs were illustrated by bioinformatic analysis. Validation of the top 3 abundant miRNAs was carried out by qRT-PCR (quantitative reverse transcription‐polymerase chain reaction). Results TEM and DLS revealed the mean size of the exosomes (RBC-derived) as 64.08 nm. These exosomes exhibited higher abundance of short RNA than the long RNA. 78 miRNAs were simultaneously detected in 3 exosome samples and mature RBCs. Several biological processes might be impacted by these miRNAs, through their target gene(s) enriched in a particular signalling pathway. The top 3 (abundant) miRNAs detected were as follows: miR-125b-5p, miR-4454, and miR-451a. qRT-PCR revealed higher abundance of miR-451a than others. Only miR-4454 and miR-451a abundance tended to increase with increasing storage time. Conclusion Exosomes derived from stored RBC units possessed multiple miRNAs and, hence, could serve various functions. The function of exosomes (RBC-derived) might be implemented partly by the predominantly enriched miR-451a.

Highlights

Allogeneic red blood cell (RBC) transfusion is an important therapeutic approach
Several studies revealed that RBC transfusion was associated with poor prognosis in some cancer types and critically ill patients or that it affected the immune system of patients who needed chronic transfusions [1,2,3,4,5,6]
Allogeneic RBC transfusion can improve the outcome in recipients by establishing blood volume, improving blood perfusion, and changing the gut microbiome [2, 18,19,20,21,22,23,24,25]

Summary

Introduction

Allogeneic red blood cell (RBC) transfusion is an important therapeutic approach. Transfusion experts suggested that transfusion-related immunomodulation (TRIM) of blood recipients might shed some light and help in understanding these phenomena [7, 8]. The presence of “something” has been reported in the blood suspension, during storage, which participated in abnormal functioning of immune cells such as T-cells and monocytes in vitro [7,8,9,10,11]. Samples drawn from the blood recipients exhibited significant abnormality in the quantity or function of cells of the immune system in vivo [3, 4, 6]. The mechanisms underlying these phenomena were not elucidated and understood clearly

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Results

Conclusion