The online version of this article (doi:10.1186/s12891-015-0652-9) contains supplementary material, which is available to authorized users.
Benjamín Fernández-Gutiérrez and José Ramón Lamas are share senior authorship.
The corresponding author declares the absence of any conflict of interest regarding the submitted manuscript. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.
PT performed the isolation of cell, culture, expansion, cytometry analysis and helped to draft the manuscript. LR carried out the miRNAs isolation from samples and adquired miRNA data. LA performed the statistical analysis of data and participated in the design of the study. MT carried out the molecular genetic studies. PLR performed the microarray statistical analysis and interpretation of results. EH performed the cell culture differentiations, histochemistry and obtained total RNA from cultures. MAG participated in the design of the study. FM and EM participated in the design of the study, performed the surgery and provided the surgical samples. BF participated in the design of the study, coordination of the study and helped to revise the manuscript. JRL performed data analysis, conceived and coordinated the study and draft the manuscript. All authors read and approved the final manuscript.
The aim of this study was to evaluate, the existence of a signature of differentially expressed microRNAs (miRNAs) during osteogenic differentiation of bone marrow MSCs from OA and healthy donors and to describe their possible implication in joint regeneration through modulation of molecular mechanisms involved in homeostatic control in OA pathophysiology.
Following phenotypic assessment of BM-MSCs obtained from OA diagnosed patients (n = 10) and non-OA (n = 10), total small RNA was isolated after osteogenic induction for 1, 10 and 21 days, miRNA profiles were generated using a commercial expression array of 754 well-characterized miRNAs. MiRNAs, with consistent differential expression were selected for further validation by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis.
A total of 246 miRNAs were differentially expressed (fold change ≥ ± 2, P ≤0.05) between OA and non-OA BM-MSC samples; these miRNAs showed variable interactions depending on the cell and differentiation status. Two miRNAs, hsa-miR-210 and hsa-miR-335-5p out of 21 used for validation showed a significant downregulated expression during induced osteogenesis. In particular hsa-miR-335-5p, a critical regulator in bone homeostasis, was further studied. hsa-miR-335-5p downregulation in OA-MSCs, as well as their host coding gene, MEST, were also assessed.
To our knowledge, this study represents the most comprehensive assessment to date of miRNA expression profiling in BM-MSCs from OA patients and their role during osteogenic differentiation. We describe the existence of a correlation between miR-335-5p expression and OA indicating the putative role of this miRNA in OA features. These findings, may contribute to our understanding of the molecular mechanisms involved in MSCs mediated homeostatic control in OA pathophysiology that could be applicable in future therapeutic approaches.
Additional file 1: Data analysis of the expression profile of miRNA sequences, from the Sanger miRBase v14 using the Taqman OpenArray MicroRNA pools A and B. (XLS 2463 kb)12891_2015_652_MOESM1_ESM.xls
Additional file 2: miRNAs showing interactions between status (OA and Control) and differentiation time points (p values <0.05 without multiple test correction applied. (TIFF 82 kb)12891_2015_652_MOESM2_ESM.tiff
Additional file 3: miRNAs without status and differentiation interaction that show differential expression between osteogenic differentiation time points (p value <0.05 without multiple test correction applied). (TIFF 123 kb)12891_2015_652_MOESM3_ESM.tiff
Additional file 4: miRNAs without status and differentiation interaction that show differential expression between OA and Control samples (p value <0.05 without multiple test correction applied). (TIFF 157 kb)12891_2015_652_MOESM4_ESM.tiff
Additional file 5: Venn diagrams showing the distribution of miRNAs analyzed and their overlapping among different status and osteogenic time points. (PDF 19 kb)12891_2015_652_MOESM5_ESM.pdf
Additional file 6: Extraction of all the experimentally validated microRNA-target interactions for hsa-miR-210 and hsa-miR-335-5p from the miRTarBase database. An extraction of miR-335 target genes related to the WNT pathway was also included. These belong to the Wnt signaling pathway, according to the enriched annotations provided in the Kyoto Encyclopedia of Genes and Genomes (KEGG) 04310. (XLS 2121 kb)12891_2015_652_MOESM6_ESM.xls
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- Signature of microRNA expression during osteogenic differentiation of bone marrow MSCs reveals a putative role of miR-335-5p in osteoarthritis
Manuel A. González
José Ramón Lamas
- BioMed Central
Neu im Fachgebiet Orthopädie und Unfallchirurgie
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