Exosomal miRNA Signatures for Late-Onset Acute Graft-Versus-Host Disease in Allogenic Hematopoietic Stem Cell Transplantation

Abstract

Recent studies have demonstrated that exosomal microRNAs (miRNAs) have the potential of facilitating molecular diagnosis. Currently, little is known about the underlying mechanism behind late-onset acute graft-versus-host disease (LA GVHD). Identifying differentially expressed miRNAs in exosomes should be useful for understanding the role of miRNAs in this disease. This study was established to investigate the relevance of miRNAs in exosomes derived from patients developing LA GVHD after allogeneic hematopoietic stem cell transplantation (HSCT). Plasma samples were collected from patients with LA GVHD (n = 5), non-GVHD (n = 5), and controls (n = 8) for exosomal miRNA expression profiling using a TaqMan low-density array; the results were validated by quantitative reverse transcription polymerase chain reaction (RT-PCR). We analyzed exosomal miRNAs differentially expressed among these three groups. MirTarBase was employed to predict potential target genes of the miRNAs specific for LA GVHD. We detected 55 miRNAs that were differentially expressed with a significant change >2.0-fold between LA GVHD and non-GVHD. Of these, we selected the 10 miRNAs (miR-423-5p, miR-19a, miR-142-3p, miR-128, miR-193b, miR-30c, miR-193a, miR-191, miR-125b, and miR-574-3p) with the most significant differential expression. Using quantitative RT-PCR, we further identified that miR-128 was significantly upregulated at the onset of LA GVHD compared with that in normal controls and is a promising diagnostic marker of LA GVHD, with an area under the curve (AUC) value of 0.975. MirTarBase analysis revealed that the predicted target genes of miR-128 are involved in the immune system and inflammation. Increased expression of miR-128 may serve as a novel, noninvasive biomarker for early LA GVHD diagnosis.

Downregulation of extracellular vesicle microRNA‑101 derived from bone marrow mesenchymal stromal cells in myelodysplastic syndrome with disease progression

“…EV fraction was purified with Exoquick-TC regent (System Biosciences) according to the manufacturer's instructions. EVs were quantitated with a nanoparticle tracking analysis (NanoSight LM10; Malven) (16)(17)(18). Isolation of BM-MSC-derived EV-miRs or cellular-miR and serum EV-miR was performed using the miRNeasy kit (Qiagen).…”

“…Screening of candidate miRNA. miRNA profiling was carried out using a TaqMan low-density miRNA array (TLDA; Thermo Fisher Scientific, Inc.), as previously reported (16,17). PCR was done on a Applied Biosystem 7900 HT thermocycler (Carlsberg) according to the manufacturer's recommended program (the reaction was first incubated at 95˚C for 2 min, followed by 50 cycles of 95˚C for 15 sec and 60˚C for 1 min) using SDS2.2 software and Data Assist (Thermo Fisher Sciences, Inc.).…”

“…Data analysis was carried out using GeneSpring software (Agilent Technologies). The Benjamin-Hochberg algorithm was used for the estimation of false discovery rates, as previously reported (16,17).…”

MicroRNAs and exosomes: Small molecules with big actions in multiple myeloma pathogenesis

“…Exosomes are lipid‐membrane vesicles which are created by endocytosis and are known as other factors that are involved in the pathogenesis of MM. It is revealed that exosomal miRNA signatures are very important in allogeneic stem cell transplantation . Exosomes are released into the extracellular milieu when the multivesicular bodies (MVBs) joined the plasma membrane .…”

“…It is revealed that exosomal miRNA signatures are very important in allogeneic stem cell transplantation. 13 Exosomes are released into the extracellular milieu when the multivesicular bodies (MVBs) joined the plasma membrane. 14 Exosomes secretion, by tumor or normal cells, including epithelial, mast, B, T, and dendritic cells, can be induced or spontaneous, and can take place under pathological and physiological situations.…”