FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2340981356> ?p ?o ?g. }

• W2340981356 endingPage "742" @default.
• W2340981356 startingPage "729" @default.
• W2340981356 abstract "•OM-MSCs share 64% miRNA homology to BM-MSCs and differentially express 26 miRNAs•CXCL12 promotes CNS myelination and is negatively regulated by miR-140-5p in BM-MSCs•miR-146a-5p negatively regulates IL-6, IL-8, TLR2, and TLR4 on OM-MSCs•These properties make OM-MSCs a suitable candidate for transplant-mediated CNS repair 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. We focused on miR-146a-5p and miR-140-5p due to their reported role in the regulation of chemokine production and myelination. The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs. Addition of CXCL12 and its pharmacological inhibitors to neural co-cultures supported these data. Studies on related miR-146a-5p targets demonstrated that OM-MSCs had lower levels of Toll-like receptors and secreted less pro-inflammatory cytokines, IL-6, IL-8, and CCL2. OM-MSCs polarized microglia to an anti-inflammatory phenotype, illustrating potential differences in their inflammatory response. Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo. 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. We focused on miR-146a-5p and miR-140-5p due to their reported role in the regulation of chemokine production and myelination. The lower expression of miR-140-5p in OM-MSCs correlated with higher secretion of CXCL12 compared with BM-MSCs. Addition of CXCL12 and its pharmacological inhibitors to neural co-cultures supported these data. Studies on related miR-146a-5p targets demonstrated that OM-MSCs had lower levels of Toll-like receptors and secreted less pro-inflammatory cytokines, IL-6, IL-8, and CCL2. OM-MSCs polarized microglia to an anti-inflammatory phenotype, illustrating potential differences in their inflammatory response. Nestin-positive OM-MSCs could therefore offer a cell transplantation alternative for CNS repair, should these biological behaviors be translated in vivo. Bone-marrow-derived mesenchymal stromal cells (BM-MSCs) have been reported to secrete neurotrophic cytokines as well as a range of pro-inflammatory cytokines, chemokines, and other soluble growth factors (Nakano et al., 2010Nakano N. Nakai Y. Seo T.B. Yamada Y. Ohno T. Yamanaka A. Nagai Y. Fukushima M. Suzuki Y. Nakatani T. Ide C. Characterization of conditioned medium of cultured bone marrow stromal cells.Neurosci. Lett. 2010; 483: 57-61Crossref PubMed Scopus (95) Google Scholar, Ma et al., 2013Ma X.L. Liu K.D. Li F.C. Jiang X.M. Jiang L. Li H.L. Human mesenchymal stem cells increases expression of a-tubulin and angiopoietin 1 and 2 in focal cerebral ischemia and reperfusion.Curr. Neurovasc. Res. 2013; 10: 103-111Crossref PubMed Scopus (41) Google Scholar). Because of these properties, their feasibility and safety of administration were assessed in clinical trials for the treatment of multiple sclerosis (MS), with results suggesting they can modulate the immune response, protect neurons from degeneration, and improve disease progression (Bonab et al., 2012Bonab M.M. Sahraian M.A. Nikbin B. Lotfi J. Khorramnia S. Motamed M.R. Togha M. Harirchian M.H. Moghadam N.B. Alikhani K. et al.Autologous mesenchymal stem cell therapy in progressive multiple sclerosis: an open label study.Curr. Stem Cell. Res. Ther. 2012; 7: 407-414Crossref PubMed Scopus (132) Google Scholar, Connick et al., 2012Connick P. Kolappan M. Crawley C. Webber D.J. Patani R. Michell A.W. Du M.Q. Luan S.L. Altmann D.R. Thompson A.J. et al.Autologous mesenchymal stem cells for the treatment of secondary progressive multiple sclerosis: an open-label phase 2a proof-of-concept study.Lancet Neurol. 2012; 11: 150-156Abstract Full Text Full Text PDF PubMed Scopus (454) Google Scholar). MSCs have been isolated from a range of tissues including bone marrow, adipose, pancreas, skin, muscle, tendon, umbilical cord, skin, and dental pulp (Hass et al., 2011Hass R. Kasper C. Böhm S. Jacobs R. Different populations and sources of human mesenchymal stem cells (MSC): a comparison of adult and neonatal tissue-derived MSC.Cell Commun. Signal. 2011; 9: 12Crossref PubMed Scopus (1099) Google Scholar, Wang et al., 2013Wang X. Wang Y. Gou W. Lu Q. Peng J. Lu S. Role of mesenchymal stem cells in bone regeneration and fracture repair: a review.Int. Orthop. 2013; 37: 2491-2498Crossref PubMed Scopus (199) Google Scholar). We have recently isolated MSCs from the lamina propria of human olfactory mucosa (OM) (termed OM-MSCs; Lindsay et al., 2013Lindsay S.L. Johnstone S.A. Mountford J.C. Sheikh S. Allan D.B. Clark L. Barnett S.C. Human mesenchymal stem cells isolated from olfactory biopsies but not bone enhance CNS myelination in vitro.Glia. 2013; 61: 368-382Crossref PubMed Scopus (53) Google Scholar); a tissue of fundamental interest in the context of neuroprotection and repair because of its ability to continually support neurogenesis throughout life (Graziadei and Monti Graziadei, 1985Graziadei P.P. Monti Graziadei G. Neurogenesis and plasticity of the olfactory sensory neurons.Ann. N. Y. Acad. Sci. 1985; 457: 127-142Crossref PubMed Scopus (86) Google Scholar). Our previous studies have demonstrated that OM-MSCs have a similar antigenic profile and differentiation properties to BM-MSCs (Lindsay et al., 2013Lindsay S.L. Johnstone S.A. Mountford J.C. Sheikh S. Allan D.B. Clark L. Barnett S.C. Human mesenchymal stem cells isolated from olfactory biopsies but not bone enhance CNS myelination in vitro.Glia. 2013; 61: 368-382Crossref PubMed Scopus (53) Google Scholar). However, the entire OM-MSC population expressed nestin, while conversely around 50% of BM-MSCs were nestin-positive, despite being isolated using identical methodology (Johnstone et al., 2015Johnstone S.A. Liley M. Dalby M.J. Barnett S.C. Comparison of human olfactory and skeletal MSCs using osteogenic nanotopography to demonstrate bone-specific bioactivity of the surfaces.Acta Biomater. 2015; 13: 266-276Crossref PubMed Scopus (16) Google Scholar). Importantly, there was a major difference in the ability of OM-MSCs to promote CNS myelination in vitro via a secreted factor(s) (Lindsay et al., 2013Lindsay S.L. Johnstone S.A. Mountford J.C. Sheikh S. Allan D.B. Clark L. Barnett S.C. Human mesenchymal stem cells isolated from olfactory biopsies but not bone enhance CNS myelination in vitro.Glia. 2013; 61: 368-382Crossref PubMed Scopus (53) Google Scholar). Since these properties can be explained by a vast number of genes, we decided to use a microRNA (miRNA) array approach to identify differences and similarities between the two MSCs. miRNAs are an abundantly expressed family of small post-transcriptional regulators (18–24 nucleotides). They control gene expression by modulating the translation (usually by repression), stability, and localization of specific mRNA targets (Ambros, 2001Ambros V. microRNAs: tiny regulators with great potential.Cell. 2001; 107: 823-826Abstract Full Text Full Text PDF PubMed Scopus (1437) Google Scholar). They regulate numerous functions ranging from cell differentiation, proliferation, and apoptosis to fat metabolism (Skalnikova et al., 2011Skalnikova H. Motlik J. Gadher S.J. Kovarova H. Mapping of the secretome of primary isolates of mammalian cells, stem cells and derived cell lines.Proteomics. 2011; 11: 691-708Crossref PubMed Scopus (161) Google Scholar). miRNAs are thought to act as regulatory signals for maintaining stemness, self-renewal, and differentiation in adult stem cells and are therefore important in controlling classic stem cell properties (Collino et al., 2011Collino F. Bruno S. Deregibus M.C. Tetta C. Camussi G. MicroRNAs and mesenchymal stem cells.Vitam. Horm. 2011; 87: 291-320Crossref PubMed Scopus (41) Google Scholar, Tomé et al., 2011Tomé M. Lopez-Romero P. Albo C. Sepulveda J.C. Fernandez-Gutierrez B. Dopazo A. Bernad A. González M.A. miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells.Cell Death Differ. 2011; 18: 985-995Crossref PubMed Scopus (232) Google Scholar). Characterization of miRNAs from MSCs of different tissue sources could be relevant not only as a marker of the cell but also to fully understand their biological activities and give an insight into what makes them different (Collino et al., 2011Collino F. Bruno S. Deregibus M.C. Tetta C. Camussi G. MicroRNAs and mesenchymal stem cells.Vitam. Horm. 2011; 87: 291-320Crossref PubMed Scopus (41) Google Scholar). Recent work has described nestin-positive MSCs as a subpopulation (Tondreau et al., 2004Tondreau T. Lagneaux L. Dejeneffe M. Massy M. Mortier C. Delforge A. Bron D. Bone, marrow-derived mesenchymal stem cells already express specific neural proteins before any differentiation.Differentiation. 2004; 72: 319-326Crossref PubMed Scopus (281) Google Scholar, Wiese et al., 2004Wiese C. Rolletschek A. Kania G. Rolletschek A. Kania G. Blyszczuk P. Tarasov K.V. Tarasova Y. Wersto R.P. Boheler K.R. et al.Nestin expression-a property of multi-lineage progenitor cells?.Cell Mol. Life Sci. 2004; 61: 2510-2522Crossref PubMed Scopus (547) Google Scholar) that originates not from the mesoderm but from the neural crest giving nestin-positive MSCs specialized niche functions over nestin-negative MSCs (Isern and Méndez-Ferrer, 2011Isern J. Méndez-Ferrer S. Stem cell interactions in a bone marrow niche.Curr. Osteoporos. Rep. 2011; 9: 210-218Crossref PubMed Scopus (46) Google Scholar, Isern et al., 2014Isern J. García-García A. Martín A.M. Arranz L. Martín-Pérez D. Torroja C. Sánchez-Cabo F. Méndez-Ferrer S. The neural crest is a source of mesenchymal stem cells with specialized hematopoietic stem cell niche function.eLife. 2014; 3: e03696Crossref Scopus (203) Google Scholar). Therefore, in this investigation, we have compared the miRNA profile of nestin-positive OM-MSCs with classical BM-MSCs to determine any important biological differences that, in particular, may be relevant to their role in cell transplantation therapies for the treatment of demyelinating conditions, such as MS. Analysis revealed 195 mature miRNAs detected in OM-MSCs (n = 4 patient samples) and BM-MSCs (n = 4 patient samples), of which 125 were equivalently expressed (EE). This demonstrates 64% identity, despite being isolated from different tissues; moreover, 27 of these EE miRNAs have already been reported for BM-MSCs (Gao et al., 2011Gao J. Yang T. Han J. Yan K. Qiu X. Zhou Y. Fan Q. Ma B. MicroRNA expression during osteogenic differentiation of human multipotent mesenchymal stromal cells from bone marrow.J. Cell Biochem. 2011; 112: 1844-1856Crossref PubMed Scopus (166) Google Scholar; Figure 1A ). These data suggest that MSCs derived from OM express a similar core set of miRNAs compared with BM. In contrast, 26 were differentially expressed (DE) across all samples, with 16 being downregulated in OM-relative to BM-MSCs (Figure 1B). These miRNAs have over 300 targets, therefore a contextual approach was adopted whereby miRNAs associated with MSC biology were identified. Since our previous comparative studies identified differences in cell proliferation, cell survival, and myelination (Lindsay et al., 2013Lindsay S.L. Johnstone S.A. Mountford J.C. Sheikh S. Allan D.B. Clark L. Barnett S.C. Human mesenchymal stem cells isolated from olfactory biopsies but not bone enhance CNS myelination in vitro.Glia. 2013; 61: 368-382Crossref PubMed Scopus (53) Google Scholar), we focused on miR-140-5p and miR-146a-5p (Figure 1B), which have already been identified as key regulators of these processes (Suzuki et al., 2010Suzuki Y. Kim H.W. Ashraf M. Haider H.K. Diazoxide potentiates mesenchymal stemcell survival via NF-kappaB-dependent miR-146a expression by targeting.Fas. Am. J. Physiol. Heart Circ. Physiol. 2010; 299: H1077-H1082Crossref PubMed Scopus (73) Google Scholar, Göttle et al., 2010Göttle P. Kremer D. Jander S. Odemis V. Engele J. Hartung H.P. Küry P. Activation of CXCR7 receptor promotes oligodendroglial cell maturation.Ann. Neurol. 2010; 68: 915-924Crossref PubMed Scopus (66) Google Scholar). qPCR confirmed a significant 3.01-fold higher expression of miR-140-5p in BM-MSCs compared with OM-MSCs (n = 4 patient samples for both, p < 0.05; Figures 1C and 1E), and a significant 7.99-fold higher expression of miR-146a-5p in OM-compared with BM-MSCs (n = 4 patient samples for both, p < 0.05; Figures 1D and 1E), confirming the miRNA analysis. GeneGo MetaCore analysis revealing high-confidence mRNA targets for miR-140-5p suggested SDF-1 (referred to as CXCL12 hereafter) secretion may be differentially regulated between the MSCs. Multiplex analysis of the conditioned media (CM) collected from OM- and BM-MSCs was performed with fibroblast (FB) and CD271-FT-CM used as comparisons (CM derived from n = 4 patient samples). Since we have previously shown that both FB-CM and CD271-FT-CM do not promote myelination (Lindsay et al., 2013Lindsay S.L. Johnstone S.A. Mountford J.C. Sheikh S. Allan D.B. Clark L. Barnett S.C. Human mesenchymal stem cells isolated from olfactory biopsies but not bone enhance CNS myelination in vitro.Glia. 2013; 61: 368-382Crossref PubMed Scopus (53) Google Scholar), these were considered an appropriate comparison to ensure secreted factors were specifically generated from OM-MSCs (see Table 1); 13 were not detected and nine were secreted to equivalent levels across all groups (CCL1, CCL2, CCL3, CCL8, CX3CL1, G-CSF, CXCL10, SCF, and VEGF). CCL11 was significantly higher within both OM-MSC-CM and CD271-FT-CM compared with BM-MSC-CM and FB-CM (p < 0.05, all comparisons), suggesting the expression of a tissue-specific chemokine rather than MSC specific. CCL13 was significantly lower within FB-CM compared with all other cell type CM (p < 0.05, all comparisons), however, it was equivalently expressed within OM-MSCs, BM-MSCs, and CD271-FT-CM. CCL5 was markedly increased in CD271-FT-CM (p < 0.01). CXCL12 was the only cytokine present in significantly greater quantities in OM-MSC-CM compared with either BM-MSC-CM (p < 0.01), CD271-FT-CM (p < 0.01), or FB-CM (p < 0.05, Figure 2A ). In addition, the neurotrophic factors BDNF, NT3, NT4/5, and NGF were assayed in BM- and OM-MSC-CM (n = 3 patient samples for both). NT3 was undetectable, and both cell types secreted equivalent low levels of BDNF (OM-MSC-CM, 19.2 ± 5.3 pg/ml; BM-MSC-CM, 11.9 ± 4.83 pg/ml) and NT4/5 (OM-MSC-CM, 21.9 ± 0.3 pg/ml; BM-MSC-CM, 20.0 ± 0.7 pg/ml). OM-MSC-CM contained significantly higher levels of NGF (33.8 ± 6.8 pg/ml) compared with BM-MSC-CM (1.2 ± 0.6 pg/ml).Table 1Multiplex Chemokine Analysis of CM Showing the Comparative Differences in Secreted CytokinesCytokineSignificant DifferenceCXCL12∗∗<OM onlyCCL1NSCCL2NSCCL3NSCCL8NSCX3CL1NSG-CSFNSCXCL10NSSCFNSVEGFNSCCL11∗<OM∗<CD271-FTCCL13∗>FBCCL5∗∗<CD271-FTNS, not significantly different; ∗<OM, ∗<CD271-FT, represents that the cytokine was secreted in significantly higher amounts in CM from OM-MSCs and CD271-FTl ∗>FB, represents it was present in significantly less amounts in fibroblasts (FB), ∗∗<CD271-FT or significantly higher in CD271-FT-CM. CXCL12 was the only chemokine secreted at significantly higher amounts in OM-MSC-CM compared with all other cell types (∗p < 0.05, ∗∗p < 0.01, n = 3 for all, as determined by one-way ANOVA, Tukey's multiple comparison). Open table in a new tab NS, not significantly different; ∗<OM, ∗<CD271-FT, represents that the cytokine was secreted in significantly higher amounts in CM from OM-MSCs and CD271-FTl ∗>FB, represents it was present in significantly less amounts in fibroblasts (FB), ∗∗<CD271-FT or significantly higher in CD271-FT-CM. CXCL12 was the only chemokine secreted at significantly higher amounts in OM-MSC-CM compared with all other cell types (∗p < 0.05, ∗∗p < 0.01, n = 3 for all, as determined by one-way ANOVA, Tukey's multiple comparison). OM- and BM-MSCs (n = 3, patient samples for both) were transfected with miR-140-5p antagomir or mimic, a random sequence miRNA molecule (scrambled control) or dH2O (Figures 2B and 2C) to confirm their ability to modulate miR-140-5p expression. MiR-140-5p antagomir silenced expression, while the mimic significantly upregulated miR-140-5p in both OM- and BM-MSCs. These data confirm that levels of miR-140-5p can be modulated by both the antagomir and mimic. To validate direct correlation of miR-140-5p and CXCL12 expression, both MSC types were transfected with the miR-140-5p antagomir or mimic, and CXCL12 mRNA levels were quantified (Figures 2D and 2E). The mimic resulted in virtually undetectable levels of CXCL12 in both MSC types. Antagomir induced a significant increase in BM-MSCs compared with control levels (p < 0.05; Figure 2E), and although the levels of CXCL12 mRNA were increased in OM-MSCs compared with the mimic, they were still below that of control OM-MSCs (Figure 2D), which inherently expressed higher levels of CXCL12 mRNA than BM-MSCs. This confirms that increased levels of miR-140-5p negatively regulate the expression of CXCL12. Our previous data suggest that OM-MSCs promoted in vitro CNS myelination via a secreted factor. Here we provide evidence that CXCL12 could be this factor. Myelinating CNS co-cultures were treated with CXCL12 (100 ng/ml), CXCL12 receptor CXCR4 blocker (AMD3100), OM-MSC-CM, as well as OM-MSC-CM treated with a neutralizing antibody to CXCL12 or AMD3100 (n = 4, all treatments and four different patient samples; Figures 3A and 3B ). Media containing the CXCL12 neutralizing antibody or AMD3100 were used as controls. Exogenous CXCL12 significantly increased myelination almost 2-fold compared with controls (p < 0.05), which was blocked by AMD3100. The pro-myelinating effect of OM-MSC-CM (p < 0.05) was also reduced by AMD3100 and when treated with the neutralizing antibody to CXCL12. These data indicate that CXCL12 is at least partially responsible for the pro-myelinating effect of OM-MSCs. We next determined if modulating miR-140-5p in OM- or BM-MSCs can affect myelination in vitro. BM-MSC-CM was collected from cells transfected with the miR-140-5p antagomir and added to cultures (n = 6 patient samples; Figures 3C and 3D). Antagomir-transfected BM-MSC-CM significantly increased myelination by 1.64 ± 0.19-fold compared with control (n = 6 patient samples, p < 0.05) or that induced by control transfections (n = 6 patient samples, p < 0.01 for both). The increased myelination was brought back to levels that were not significant from control by the addition of either the antibody to CXCL12 or AMD3100 (n = 3 patient samples for both). CM derived from BM-MSCs transfected with miR-140-5p mimic (n = 3 patient samples) showed no increase in myelination from control. OM-MSC-CM collected from cells treated with miR-140-5p antagomir increased myelination compared with controls (p < 0.05) but was not significantly different to that produced by control transfections (n = 6, patient samples; Figures 3C and 3E). The pro-myelinating effect of CM collected from antagomir-treated OM-MSCs could be reduced in the presence of either the antibody to CXCL12 or AMD3100 (n = 3 patient samples for both). OM-MSC-CM collected from miR-140-5p mimic-treated cells reduced myelination to control. This confirms that miR-140-5p can induce OM- and BM-MSCs to increase CXCL12 secretion, which is pro-myelinating, since blocking its activity abolishes the effect. CXCL12 is known to mediate its effect via CXCR4 and CXCR7, therefore cellular expression may help determine the mode of action. Oligodendrocyte precursor cells (OPCs) and microglia were both found to express CXCR4, however no expression was found on astrocytes (n = 3, all cell types; Figures 4A and 4B ). Western blotting of CXCR4 revealed OPCs (n = 6) and microglia (n = 4) to have at least three distinct isoforms, however, both had differential expression of each (Figures 4B and 4C). It was found that the most abundant isoform expressed within microglia was the 50 kDa isoform, while OPCs were found to predominantly have the 45 kDa isoform. Total protein quantification of all CXCR4 isoforms was found not to be significantly different between microglia (1.72 ± 0.31 a.u.) versus OPCs (1.33 ± 0.35 a.u.). CXCR7 expression was only barely detectable by western blot and not immunocytochemistry on both OPCs and microglia (Figure 4B). Therefore the pro-myelinating capabilities of CXCL12 could be mediated via its action on OPCs or microglia, or both. Purified OPCs were treated with CXCL12, OM-, or BM-MSC-CM (from three different patient samples for both) and labeled with markers of OPC differentiation (n = 3; Figure 4D). There were no differences in immunoreactivity of NG2 (early OPC marker), O4 (middle/late OPC marker), or PLP (late myelinating OPC marker). However, CXCL12 significantly changed OPC morphological appearance from a predominantly simple (bipolar) to a more complex (multi-branched) morphology, similar to that found with OM- or BM-MSC-CM treatment (Figure 4E; p < 0.001). OM-MSC-CM resulted in significantly more OPCs exhibiting a membranous morphology when compared with either control or BM-MSC-CM (p < 0.001 for both), and although the result of OM-MSC-CM treatment looked to be greater, it was not significantly different to CXCL12 treatment. Overall this suggests that CXCL12 can mediate morphological OPC differentiation via its direct action on the OPC itself. This hypothesis was confirmed by examining the effect of CXCL12, OM-, and BM-MSC-CM on purified OPCs incubated with inert nanofibers (Figure 4F). In these experiments, PLP-positive OPCs had significantly greater cell areas ensheathing axons in both CXCL12 and OM-MSC-CM compared with controls (n = 3 each treatment and patient samples). BM-MSC-CM also caused an increase in PLP-positive cell area, however this was found not to be significantly different from control due to the variability among samples (n = 3 patient samples). This suggests that both CXCL12 and OM-MSC-CM promoted process extension and wrapping in the absence of axonal signals to a greater extent than BM-MSC-CM. Microglia are thought to polarize into distinct phenotypes, pro-inflammatory (depicted by iNOS expression) or anti-inflammatory (increased arginase I expression). To determine if the microglia phenotype plays a role in mediating myelination, we treated purified microglia with OM-, BM-MSC-CM (n = 4 patient samples for both), or CXCL12 (n = 4). CM from both cell types were tested for endogenous endotoxin levels (Pierce Endotoxin Quantification Kit; Thermo Scientific) to rule out any difference as a result of CM contamination prior to use. Lipopolysaccharide (LPS) and IL-4 were used as controls to induce the pro- and anti-inflammatory phenotype, respectively (Figures 4G and 4H). LPS stimulation shifted microglia predominantly to the pro-inflammatory phenotype assessed by increased iNOS expression compared with control (n = 4, p < 0.01), while IL-4-treated microglia expressed arginase I with no iNOS detectable (n = 4), suggesting a population that is similar to control. Treatment of microglia with OM-MSC-CM and CXCL12 increased arginase I expression compared with the control (p < 0.001 and p < 0.05, respectively) with no detectable iNOS expression, suggesting a larger shift to the anti-inflammatory phenotype in both conditions. BM-MSC-CM did not stimulate a significant increase in arginase I levels from control but did however lead to increased iNOS production (p < 0.01), the only treatment besides LPS to do so (Figures 4G and 4H). This would strongly suggest that OM-MSC-CM and CXCL12 cause microglia to polarize predominantly to an anti-inflammatory phenotype, in contrast to BM-MSC-CM, which appears to shift them more toward a pro-inflammatory phenotype. Recently, the anti-inflammatory phenotype of microglia has been reported to play a role in myelination through the production of activin A (Miron et al., 2013Miron V.E. Boyd A. Zhao J.W. Yuen T.J. Ruckh J.M. Shadrach J.L. van Wijngaarden P. Wagers A.J. Williams A. Franklin R.J. et al.M2 microglia and macrophages drive oligodendrocyte differentiation during CNS remyelination.Nat. Neurosci. 2013; 16: 1211-1218Crossref PubMed Scopus (1107) Google Scholar). Therefore we assessed activin A levels within the CM of microglia treated with CXCL12 (n = 3) or OM- or BM-MSC-CM (n = 3 patient samples for both) with their respective CM used as controls (Figure 4I). It was found that there were no identifiable increases in activin A in CXCL12-treated cultures, and although OM- and BM-MSC-CM showed increased levels, this was similar to that already present within the CM alone. Therefore, both types of MSC, but not microglia, secrete activin A. The GeneGo MetaCore network for miR-146a-5p indicated an association with Toll-like receptor (TLR) expression and the regulation of inflammatory chemokines. This suggests differential modulation of the inflammatory response, which is an important consideration for cell transplant-mediated repair. Total TLR2 and 4 levels were significantly less in OM-MSCs compared with BM-MSCs (n = 6 patient samples for both, p < 0.01 and p < 0.05, respectively; Figure 5A ). There was notably less TLR2 expression in both cell types compared with their TLR4 expression since only TLR4 was present abundantly enough to be detected via fluorescence-activated cell sorting (FACS) (n = 3 patient samples for both; Figure 5B). MiR-146a-5p is thought to modulate the secretion of IL-6 and IL-8, and BM-MSC-CM was found to contain at least 1.5-fold more of both these and CCL2 than OM-MSC-CM (Figure 5C). Quantification was carried out by ELISA (n = 3 patient samples for both, p < 0.01, p < 0.001, and p < 0.05; Figure 5D). 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. In this investigation, miRNA-based fingerprinting demonstrated that OM- and BM-MSCs were 64% homologous, suggesting they have related regulatory miRNA patterns. Others have shown that MSCs derived from different tissue niches share expression of a core set of miRNAs that regulate associated target genes, although miRNA similarity between the two MSC types here was greater than that reported for other MSCs (Lazzarini et al., 2014Lazzarini R. Olivieri F. Ferretti C. Mattioli-Belmonte M. Di Primio R. Orciani M. mRNAs and miRNAs profiling of mesenchymal stem cells derived from amniotic fluid and skin: the double face of the coin.Cell Tissue Res. 2014; 355: 121-130Crossref PubMed Scopus (28) Google Scholar). We identified 26 DE miRNAs, which could explain their different biological properties. We focused on miRNAs associated with chemokine production and myelination. and therefore examined miR-146a-5p, and miR-140-5p in more detail due to their reported association (Nicolas et al., 2008Nicolas F.E. Pais H. Schwach F. Lindow M. Kauppinen S. Moulton V. Dalmay T. Experimental identification of microRNA-140 targets by silencing and overexpressing miR-140.RNA. 2008; 14: 2513-2520Crossref PubMed Scopus (99) Google Scholar, Suzuki et al., 2010Suzuki Y. Kim H.W. Ashraf M. Haider H.K. Diazoxide potentiates mesenchymal stemcell survival via NF-kappaB-dependent miR-146a expression by targeting.Fas. Am. J. Physiol. Heart Circ. Physiol. 2010; 299: H1077-H1082Crossref PubMed Scopus (73) Google Scholar, Göttle et al., 2010Göttle P. Kremer D. Jander S. Odemis V. Engele J. Hartung H.P. Küry P. Activation of CXCR7 receptor promotes oligodendroglial cell maturation.Ann. Neurol. 2010; 68: 915-924Crossref PubMed Scopus (66) Google Scholar, Hsieh et al., 2013Hsieh J.Y. Huang T.S. Cheng S.M. Lin W.S. Tsai T.N. Lee O.K." @default.
• W2340981356 created "2016-06-24" @default.
• W2340981356 creator A5012713262 @default.
• W2340981356 creator A5029491830 @default.
• W2340981356 creator A5031478913 @default.
• W2340981356 creator A5074019603 @default.
• W2340981356 creator A5089974683 @default.
• W2340981356 date "2016-05-01" @default.
• W2340981356 modified "2023-10-15" @default.
• W2340981356 title "Comparative miRNA-Based Fingerprinting Reveals Biological Differences in Human Olfactory Mucosa- and Bone-Marrow-Derived Mesenchymal Stromal Cells" @default.
• W2340981356 cites W14098964 @default.
• W2340981356 cites W1962971401 @default.
• W2340981356 cites W1966940022 @default.
• W2340981356 cites W1969119054 @default.
• W2340981356 cites W1970404807 @default.
• W2340981356 cites W1977213877 @default.
• W2340981356 cites W1984008481 @default.
• W2340981356 cites W1990460115 @default.
• W2340981356 cites W1991677352 @default.
• W2340981356 cites W1993729408 @default.
• W2340981356 cites W1996835079 @default.
• W2340981356 cites W1998225973 @default.
• W2340981356 cites W2000804908 @default.
• W2340981356 cites W2004375259 @default.
• W2340981356 cites W2005349078 @default.
• W2340981356 cites W2012056552 @default.
• W2340981356 cites W2035718667 @default.
• W2340981356 cites W2038651295 @default.
• W2340981356 cites W2039611035 @default.
• W2340981356 cites W2043834555 @default.
• W2340981356 cites W2047175748 @default.
• W2340981356 cites W2068376754 @default.
• W2340981356 cites W2073527066 @default.
• W2340981356 cites W2075860675 @default.
• W2340981356 cites W2080091183 @default.
• W2340981356 cites W2105051867 @default.
• W2340981356 cites W2109321140 @default.
• W2340981356 cites W2116748550 @default.
• W2340981356 cites W2117898848 @default.
• W2340981356 cites W2123954788 @default.
• W2340981356 cites W2125759758 @default.
• W2340981356 cites W2130571779 @default.
• W2340981356 cites W2133112122 @default.
• W2340981356 cites W2138093663 @default.
• W2340981356 cites W2153949260 @default.
• W2340981356 cites W2155937459 @default.
• W2340981356 cites W2159560463 @default.
• W2340981356 cites W2164485217 @default.
• W2340981356 cites W2169859745 @default.
• W2340981356 cites W2171069275 @default.
• W2340981356 cites W2221064544 @default.
• W2340981356 cites W54166946 @default.
• W2340981356 doi "https://doi.org/10.1016/j.stemcr.2016.03.009" @default.
• W2340981356 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4940454" @default.
• W2340981356 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/27117785" @default.
• W2340981356 hasPublicationYear "2016" @default.
• W2340981356 type Work @default.
• W2340981356 sameAs 2340981356 @default.
• W2340981356 citedByCount "23" @default.
• W2340981356 countsByYear W23409813562017 @default.
• W2340981356 countsByYear W23409813562018 @default.
• W2340981356 countsByYear W23409813562019 @default.
• W2340981356 countsByYear W23409813562020 @default.
• W2340981356 countsByYear W23409813562021 @default.
• W2340981356 countsByYear W23409813562022 @default.
• W2340981356 countsByYear W23409813562023 @default.
• W2340981356 crossrefType "journal-article" @default.
• W2340981356 hasAuthorship W2340981356A5012713262 @default.
• W2340981356 hasAuthorship W2340981356A5029491830 @default.
• W2340981356 hasAuthorship W2340981356A5031478913 @default.
• W2340981356 hasAuthorship W2340981356A5074019603 @default.
• W2340981356 hasAuthorship W2340981356A5089974683 @default.
• W2340981356 hasBestOaLocation W23409813561 @default.
• W2340981356 hasConcept C104317684 @default.
• W2340981356 hasConcept C142724271 @default.
• W2340981356 hasConcept C145059251 @default.
• W2340981356 hasConcept C16930146 @default.
• W2340981356 hasConcept C169760540 @default.
• W2340981356 hasConcept C198826908 @default.
• W2340981356 hasConcept C201792869 @default.
• W2340981356 hasConcept C202751555 @default.
• W2340981356 hasConcept C203014093 @default.
• W2340981356 hasConcept C2780007613 @default.
• W2340981356 hasConcept C2781377823 @default.
• W2340981356 hasConcept C2993373878 @default.
• W2340981356 hasConcept C502942594 @default.
• W2340981356 hasConcept C54355233 @default.
• W2340981356 hasConcept C71924100 @default.
• W2340981356 hasConcept C86803240 @default.
• W2340981356 hasConcept C95444343 @default.
• W2340981356 hasConceptScore W2340981356C104317684 @default.
• W2340981356 hasConceptScore W2340981356C142724271 @default.
• W2340981356 hasConceptScore W2340981356C145059251 @default.
• W2340981356 hasConceptScore W2340981356C16930146 @default.
• W2340981356 hasConceptScore W2340981356C169760540 @default.
• W2340981356 hasConceptScore W2340981356C198826908 @default.
• W2340981356 hasConceptScore W2340981356C201792869 @default.
• W2340981356 hasConceptScore W2340981356C202751555 @default.

• next