Serum and Extracellular Vesicle Micrornas MiR-423, MiR-199 and MiR-93* As Biomarkers for Acute Graft Versus Host Disease

Abstract

Acute graft-versus-host disease (aGvHD) is a major cause of adverse outcome in hematopoietic stem cell transplantation (HSCT), with a high incidence (20-50%). A novel, non-invasive diagnostic test to predict for prevalence and severity would enable improved prophylaxis and reduce morbidity. Circulatory microRNAs (miRNAs) miR-423, miR-199, miR-93*, and miR-377 have previously been associated with aGvHD in post-HSCT patient plasma, but validation is lacking and their expression within extracellular vesicles (EVs) has not been explored. This study replicated elevated serum expression of miR-423 (p < 0.001), miR-199 (p = 0.04), miR-93* (p < 0.001), and miR-377 (p = 0.03) in aGvHD, using a prognostic cohort of day 14 (D14) post-HSCT patient samples (n = 81). Expression also associated with disease severity. Further analysis at aGvHD diagnosis in an independent cohort (n = 65) confirmed high miR-423 (p = 0.02), miR-199 (p = 0.007), and miR-93* (p = 0.004) expression at disease onset. Investigation of expression patterns during early HSCT sequential timepoints (pre-HSCT to D28) identified elevated miRNAs at D7 post-HSCT in all transplant patients. In a novel investigation of miRNA expression in serum EVs (n = 15), miR-423 (p = 0.09), miR-199 (p = 0.008), and miR-93* (p = 0.001) levels were lower at D14 in patients who later developed aGvHD, and this was replicated for miR-423 (p = 0.02) and miR-199 (p = 0.04) (n = 47). Comparing serum to circulating EVs, at D14 patients remaining aGvHD-free had higher expression of miR-423 (p = 0.03), miR-199 (p = 0.009), and miR-93* (p = 0.002) in the EV fraction. Results verify the capacity for circulating miR-423, miR-199, and miR-93* as diagnostic and prognostic aGvHD biomarkers. The novel finding of their differential expression in EVs suggests a potential role in aGvHD etiology. KeY POinTs• Expression of miR-423, miR-199, and miR-93* in post-HSCT serum and extracellular vesicles is associated with aGvHD incidence and severity. • Differential expression in extracellular vesicles suggests these microRNAs may be implicated in the pathology of aGvHD.

Development of acoustically isolated extracellular plasma vesicles for biomarker discovery in allogeneic hematopoietic stem cell transplantation

“…It has been demonstrated that expression of miR-93*/ miR-93-3p together with miR-423, miR-199a-3p and miR-377 in the plasma was able to predict the probability of aGvHD occurrence and was furthermore positively associated with disease severity and patient survival [4]. Importantly, high expression of these miRNAs was detectable before GvHD was diagnosed clinically [4,44]. However, our data did not show a positive correlation of miR-93* with GvHD status but instead we found that miR-93-5p, which is another strand of miR-93, was negatively correlated with GvHD.…”

Potential roles of extracellular vesicles in the pathophysiology, diagnosis, and treatment of autoimmune diseases

“…In the diagnosis and treatment of graft-versus-host disease (GVHD), the application of EVs has made more advancement. The EVs isolated from patient serum contains three miRNAs (miR-423, miR-199, miR-93), which may be related to the incidence and severity of GVHD [68]; and the expression of CD146, CD31, and CD140-α on EVs surface, is also closely related to the onset of the disease [69]. Traditional MSCs therapy after replacement with EVs for GVHD also showed ideal results, suggesting that MSCs-derived EVs are potential for cell-free therapy for GVHD [58,70].…”

miRBaseMiner, a tool for investigating miRBase content

“…In earlier miRBase releases, a '*' was used to indicate the miRNA was the minor product of the precursor maturation process and not functional. However, as product from both arms has been subsequently shown to be functional [7] this suffix is gradually being retired from miRBase, although its usage continues in the literature [21][22][23][24][25][26]. Additionally, the species code has been modified over time to reflect changes in the accepted species name; for example, the species code for Capitella teleta (NCBI-taxid: 283909) was updated from cap to cte in version 15 (to reflect the change from 'Capitella sp' to 'Capitella teleta' as the commonly accepted name).…”