FIGURE 6.

Inhibition of miR‐31‐5p boosts osteogenesis of BMSCs from the aged‐vascular niche through de‐repression of Wnt‐axis. (a) Volcano plot depicting differential expression analysis of miRs in plasma from HGPS patients (n = 3) and age‐ and gender‐matched controls (n = 3) using depicted thresholds with Benjamini‐Hochberg adjusted p‐values. Gene expression levels of miR‐31‐5p in plasma from HGPS patients and unaffected controls (n = 4). (b) miR‐31‐5p gene expression levels in bmECs and enriched BMSCs derived from Wt animals (young ~3 months and aged ~22 months, n = 5–7) and (c) EC‐depleted BM (BM), BMSCs, extracellular vesicles derived from EC‐conditioned media (EC CM) and bmECs derived from Wt and Prog‐Tg animals (age ≤ 14 days (black color), age = 30–35 weeks (red color), n = 4–7). (d, e) Osteogenic differentiation of scramble (scr) and antimiR31‐5p (ant‐31) treated (d) adult BMSCs and (e) Prog‐Tg derived BMSCs with quantification of ARS‐stained deposits (age ~ 5–22 months for adult, age ≤ 14 days for Prog‐Tg; n = 5–6). (f) Gene expression analysis of Fzd3, Lef1, and Alpl shown as fold change to corresponding scramble control (age ≤ 14 days, n = 4–6). (b, c) Unpaired two‐tailed Student's t‐test (*p < 0.05, ns, not significant). (d) Paired Student's t‐test (antimiR‐31‐5p treated vs. scramble, *p < 0.05). (e, f) Wilcoxon Signed Rank test (antimiR‐31‐5p treated vs. scramble, *p < 0.05). (g) Proposed model depicting miR‐31‐mediated paracrine Wnt‐repressive signals derived from aged‐vascular niche hinder osteogenic differentiation capacity of BMSCs. Inhibition of miR‐31‐5p (antimiR‐31) boosts osteogenesis in BMSCs through de‐repression of Fzd3/Lef1‐levels. Senescent and paracrine senescent ECs marked in grey and darker red colors.