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Comparative miRNA-Based Fingerprinting Reveals Biological Differences in Human Olfactory Mucosa- and Bone-Marrow-Derived Mesenchymal Stromal Cells.

Lindsay SL, Johnstone SA, McGrath MA, Mallinson D, Barnett SC - Stem Cell Reports (2016)

Bottom Line: Previously we reported that nestin-positive human mesenchymal stromal cells (MSCs) derived from the olfactory mucosa (OM) enhanced CNS myelination in vitro to a greater extent than bone-marrow-derived MSCs (BM-MSCs). miRNA-based fingerprinting revealed the two MSCs were 64% homologous, with 26 miRNAs differentially expressed.The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs.Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Inflammation and Immunity, Glasgow Biomedical Research Centre, University of Glasgow, Sir Graeme Davies Building, 120 University Place, Glasgow G12 8TA, UK.

No MeSH data available.


Related in: MedlinePlus

Regulation of IL-6 and IL-8 by miR-146a-5p(A) qPCR of miR-146a-5p levels in OM-MSCs (n = 3 patient samples) and BM-MSCs (n = 3 patient samples) after transduction with antagomir. There was a significant reduction within OM-MSCs (mean ± SEM, ∗p < 0.05, one-way ANOVA with Tukey's multiple comparison).(B–D) Quantification of IL-8 (B), IL-6 (C), and CCL2 (D) in OM-MSC-CM (OM-MSCs, n = 4 patient samples) and BM-MSC-CM (BM-MSCs, n = 4 patient samples) before and after LPS treatment (Untransfected), and OM-MSCs after transfection with the antagomir to miR-146a-5p (Antagomir Transfected). (B) OM-MSCs secrete less IL-8 compared with BM-MSCs. LPS stimulation of OM-MSCs caused an increase in IL-8 levels, which was significantly greater in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison). (C) IL-6 levels were significantly less in OM-MSCs compared with BM-MSCs both before and after LPS stimulation. IL-6 could be induced by LPS to greater levels in the presence of the antagomir (mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p<0.001, one-way ANOVA, Tukey's multiple comparison). (D) CCL2 was secreted significantly less in OM-MSC-CM compared with BM-MSC-CM. There were no differences before or after LPS stimulation or in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
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fig6: Regulation of IL-6 and IL-8 by miR-146a-5p(A) qPCR of miR-146a-5p levels in OM-MSCs (n = 3 patient samples) and BM-MSCs (n = 3 patient samples) after transduction with antagomir. There was a significant reduction within OM-MSCs (mean ± SEM, ∗p < 0.05, one-way ANOVA with Tukey's multiple comparison).(B–D) Quantification of IL-8 (B), IL-6 (C), and CCL2 (D) in OM-MSC-CM (OM-MSCs, n = 4 patient samples) and BM-MSC-CM (BM-MSCs, n = 4 patient samples) before and after LPS treatment (Untransfected), and OM-MSCs after transfection with the antagomir to miR-146a-5p (Antagomir Transfected). (B) OM-MSCs secrete less IL-8 compared with BM-MSCs. LPS stimulation of OM-MSCs caused an increase in IL-8 levels, which was significantly greater in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison). (C) IL-6 levels were significantly less in OM-MSCs compared with BM-MSCs both before and after LPS stimulation. IL-6 could be induced by LPS to greater levels in the presence of the antagomir (mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p<0.001, one-way ANOVA, Tukey's multiple comparison). (D) CCL2 was secreted significantly less in OM-MSC-CM compared with BM-MSC-CM. There were no differences before or after LPS stimulation or in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).

Mentions: The miR-146a-5p antagomir (Figure 6A) caused a significant reduction in miR-146a-5p miRNA levels in OM-MSCs (p < 0.05), however BM-MSCs, which have less of this miRNA constitutively, showed a non-significant reduction (n = 3, patient samples for both). The levels of IL-8, IL-6, and CCL2 were all assessed before and after LPS stimulation for 24 hr in CM collected from BM-MSCs, OM-MSCs, OM-MSC transfected with dH2O or with a scrambled control, or with the miR-146-5p antagomir (n = 4 all treatments and n = 4 patient samples for both; Figures 6B–6D). Both control transfections expressed similar non-significant levels of cytokine expression before and after stimulation with LPS, therefore only dH2O-transfected OM-MSC control data are presented. The higher secretion of IL-8, IL-6, and CCL2 within BM-MSC-CM in basal conditions compared with OM-MSC-CM was confirmed (p < 0.05, p < 0.01, p < 0.05, respectively). LPS caused IL-8 to increase to equivalent levels within OM-MSC-CM and BM-MSC-CM. However, OM-MSCs in the presence of the miR-146a-5p antagomir produced a significantly larger increase in LPS-stimulated IL-8 levels (p < 0.05), suggesting that repression of miR-146a-5p led to increased production of IL-8. IL-6 levels were found to be lower in OM-MSC-CM both before and after LPS stimulation when compared with BM-MSC-CM (p < 0.01 and p < 0.001, respectively). Transfection with miR-146a-5p antagomir caused an increase in the LPS-induced levels of IL-6 in OM-MSCs (p < 0.05). CCL2 levels, although lower in OM-MSCs compared with BM-MSCs under basal conditions (p < 0.05), was found not to be significantly different after LPS stimulation in antagomir-treated OM-MSCs (Figure 6D). These data provide evidence that miR-146-5p can regulate the secretion of both IL-6 and IL-8 but not CCL2 in both OM- and BM-MSCs.


Comparative miRNA-Based Fingerprinting Reveals Biological Differences in Human Olfactory Mucosa- and Bone-Marrow-Derived Mesenchymal Stromal Cells.

Lindsay SL, Johnstone SA, McGrath MA, Mallinson D, Barnett SC - Stem Cell Reports (2016)

Regulation of IL-6 and IL-8 by miR-146a-5p(A) qPCR of miR-146a-5p levels in OM-MSCs (n = 3 patient samples) and BM-MSCs (n = 3 patient samples) after transduction with antagomir. There was a significant reduction within OM-MSCs (mean ± SEM, ∗p < 0.05, one-way ANOVA with Tukey's multiple comparison).(B–D) Quantification of IL-8 (B), IL-6 (C), and CCL2 (D) in OM-MSC-CM (OM-MSCs, n = 4 patient samples) and BM-MSC-CM (BM-MSCs, n = 4 patient samples) before and after LPS treatment (Untransfected), and OM-MSCs after transfection with the antagomir to miR-146a-5p (Antagomir Transfected). (B) OM-MSCs secrete less IL-8 compared with BM-MSCs. LPS stimulation of OM-MSCs caused an increase in IL-8 levels, which was significantly greater in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison). (C) IL-6 levels were significantly less in OM-MSCs compared with BM-MSCs both before and after LPS stimulation. IL-6 could be induced by LPS to greater levels in the presence of the antagomir (mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p<0.001, one-way ANOVA, Tukey's multiple comparison). (D) CCL2 was secreted significantly less in OM-MSC-CM compared with BM-MSC-CM. There were no differences before or after LPS stimulation or in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
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fig6: Regulation of IL-6 and IL-8 by miR-146a-5p(A) qPCR of miR-146a-5p levels in OM-MSCs (n = 3 patient samples) and BM-MSCs (n = 3 patient samples) after transduction with antagomir. There was a significant reduction within OM-MSCs (mean ± SEM, ∗p < 0.05, one-way ANOVA with Tukey's multiple comparison).(B–D) Quantification of IL-8 (B), IL-6 (C), and CCL2 (D) in OM-MSC-CM (OM-MSCs, n = 4 patient samples) and BM-MSC-CM (BM-MSCs, n = 4 patient samples) before and after LPS treatment (Untransfected), and OM-MSCs after transfection with the antagomir to miR-146a-5p (Antagomir Transfected). (B) OM-MSCs secrete less IL-8 compared with BM-MSCs. LPS stimulation of OM-MSCs caused an increase in IL-8 levels, which was significantly greater in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison). (C) IL-6 levels were significantly less in OM-MSCs compared with BM-MSCs both before and after LPS stimulation. IL-6 could be induced by LPS to greater levels in the presence of the antagomir (mean ± SEM, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p<0.001, one-way ANOVA, Tukey's multiple comparison). (D) CCL2 was secreted significantly less in OM-MSC-CM compared with BM-MSC-CM. There were no differences before or after LPS stimulation or in the presence of the antagomir (mean ± SEM, ∗p < 0.05, one-way ANOVA, Tukey's multiple comparison).
Mentions: The miR-146a-5p antagomir (Figure 6A) caused a significant reduction in miR-146a-5p miRNA levels in OM-MSCs (p < 0.05), however BM-MSCs, which have less of this miRNA constitutively, showed a non-significant reduction (n = 3, patient samples for both). The levels of IL-8, IL-6, and CCL2 were all assessed before and after LPS stimulation for 24 hr in CM collected from BM-MSCs, OM-MSCs, OM-MSC transfected with dH2O or with a scrambled control, or with the miR-146-5p antagomir (n = 4 all treatments and n = 4 patient samples for both; Figures 6B–6D). Both control transfections expressed similar non-significant levels of cytokine expression before and after stimulation with LPS, therefore only dH2O-transfected OM-MSC control data are presented. The higher secretion of IL-8, IL-6, and CCL2 within BM-MSC-CM in basal conditions compared with OM-MSC-CM was confirmed (p < 0.05, p < 0.01, p < 0.05, respectively). LPS caused IL-8 to increase to equivalent levels within OM-MSC-CM and BM-MSC-CM. However, OM-MSCs in the presence of the miR-146a-5p antagomir produced a significantly larger increase in LPS-stimulated IL-8 levels (p < 0.05), suggesting that repression of miR-146a-5p led to increased production of IL-8. IL-6 levels were found to be lower in OM-MSC-CM both before and after LPS stimulation when compared with BM-MSC-CM (p < 0.01 and p < 0.001, respectively). Transfection with miR-146a-5p antagomir caused an increase in the LPS-induced levels of IL-6 in OM-MSCs (p < 0.05). CCL2 levels, although lower in OM-MSCs compared with BM-MSCs under basal conditions (p < 0.05), was found not to be significantly different after LPS stimulation in antagomir-treated OM-MSCs (Figure 6D). These data provide evidence that miR-146-5p can regulate the secretion of both IL-6 and IL-8 but not CCL2 in both OM- and BM-MSCs.

Bottom Line: Previously we reported that nestin-positive human mesenchymal stromal cells (MSCs) derived from the olfactory mucosa (OM) enhanced CNS myelination in vitro to a greater extent than bone-marrow-derived MSCs (BM-MSCs). miRNA-based fingerprinting revealed the two MSCs were 64% homologous, with 26 miRNAs differentially expressed.The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs.Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection, Inflammation and Immunity, Glasgow Biomedical Research Centre, University of Glasgow, Sir Graeme Davies Building, 120 University Place, Glasgow G12 8TA, UK.

No MeSH data available.


Related in: MedlinePlus