NGS analysis revealed distinct miRNA Expression Profiles in Eosinophil Subtypes in Patients with Allergic Asthma

<bold>Introduction:</bold> MiRNAs are small, non-coding RNAs regulating cellular functions. Several distinct expressed miRNAs have been identified in eosinophils of allergic asthma (AA) patients, but their subtype-specific profiles that distinguish inflammatory (iEOS) from lung-resident eosinophils (rEOS), remain unknown <bold>Aim:</bold> To identify miRNA expression profile differences between iEOS-like and rEOS-like cells in AA <bold>Methods:</bold> We studied 6 AA patients and 4 healthy subjects (HS). Total blood eosinophils were isolated by high-density centrifugation and magnetic separation, phenotyping – with magnetic beads conjugated antibodies against CD62L. RNA libraries were prepared using NEXTflex Small RNA-Seq Kit V4 <bold>Results:</bold> NGS analysis revealed increased miR-23c expression in rEOS-like cells compared to iEOS-like cells in AA patients, p < 0.05, with no significant differences from HS rEOS-like cells. Also, miR-128-1-5p and miR-28-3p were upregulated in both eosinophil subtypes from AA patients compared to HS eosinophils, with higher expression in AA rEOS-like cells, p < 0.05. Furthermore, miR-3d-5p, miR-55-3p, let-7a-5p, miR-1287-5p, miR-126a, miR-45b-5p, miR-22-5p, miR-125a-5p, miR-548j-5p, let-7b-3p, miR-7-1-3p, and miR-34-3p were upregulated iEOS-like cells compared to rEOS-like cells in AA, p < 0.05, with no differences from HS iEOS-like cells. A subset of these miRNAs (miR-55-3p, miR-126a, miR-22-5p, miR-125a-5p, let-7b-3p, miR-7-1-3p, and miR-34-3p) also showed lower expression in AA rEOS-like cells compared to HS rEOS-like cells, whereas let-7a-5p was upregulated in AA rEOS-like cells compared to HS rEOS-like cells, p < 0.05 <bold>Conclusions:</bold> The identified miRNAs exhibit diverse expression patterns across eosinophil subtypes in AA, reflecting their potential roles in subtype-specific activity regulation in asthma