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Transcriptional effects of LMP3 in MSC

ABSTRACT: Lim Mineralization Protein, splice variant-3 (LMP3) is known to induce osteoblast differentiation as a result of active modulation of selected molecules involved in the osteogenic cascade. LMP3 seems to act as an upstream modulator of osteogenic differentiation, although the complete network of molecular events involved in this process are still largely unknown. In particular, LMP3 seems to act, at least in part, through the upregulation of bone morphogenetic proteins (BMPs) which are known to exert multiple functions, leading to the regulation of diverse cellular processes. Thus, LMP actions inside the cell are likely to involve the activation of main signaling pathways, suggesting a possible wider role of the molecule in cell homeostasis. The aim of this study is to analyze the gene expression profiles exhibited by human bone marrow mesenchymal stem cells (MSCs) expressing the LMP3 gene in order to investigate the early transcriptional events preceding the acquisition of a differentiated phenotype. For this purpose, MSCs were transduced using a replication-defective adenoviral vector carrying the human LMP3 gene. Gene expression profiling was then carried out by microarray analysis one day after cell transduction. The results obtained in this study suggest that in addition to activating the TGFB1-signaling pathway, LMP3 acts on the fine balance between proliferation and differentiation of mesenchymal cells. Gene modulation directly related to the reprogramming of the transcriptional machinery induced by the viral vector is also discussed.

ORGANISM(S): Homo sapiens

PROVIDER: GSE16614 | GEO | 2010/12/31

SECONDARY ACCESSION(S): PRJNA116253

REPOSITORIES: GEO

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Project description:Human mesenchymal stem cells (hMSC) have an extensive potential for clinical applications in cell therapy. However, very little is known of the specific molecular regulatory mechanisms that control the therapeutical properties of these cells. We aimed to identify microRNAs (miRNAs) that could be involved in controlling the transition between the self-renewing (undifferentiated) and the reparative (differentiated) phenotypes of hMSCs. MicroRNA microarrays were used to identify miRNAs that are upregulated in undifferentiated hMSCs. For that, we compared the miRNA expression profiles of undifferentiated bone marrow-derived hMSCs with the same primary cell lines after 9 days of in vitro adipogenic or osteogenic induction. We also compared the miRNA expression profiles of undifferentiated hMSCs with skin fibroblasts (a mesenchymal cell lineage with a more restricted differentiation potential). These experiments allowed us to identify miR-335 as the only miRNA downregulated upon MSC differentiation as well as in MSCs in comparison with skin fibroblasts. Gene expression microarrays were used to identify genes that are downregulated in hMSCs overexpressing miR-335. We compared the miRNA expression profiles of hMSCs transduced with a lentiviral vector encoding miR-335 with MSCs transduced with a control lentiviral vector. Our results suggest miR-335 downregulation could be one of the triggers for the initiation of MSCs activities involved in tissue repair and remodelling, including cell migration and differentiation. We compared the miRNA expression profiles of undifferentiated bone marrow-derived hMSCs with the same primary cell lines after 9 days of adipogenic or osteogenic induction, as well as with skin fibroblasts. A total of four independent samples were used for each condition. For the adipogenic/osteogenic vs. undifferentiated MSC comparison, the RNA samples were pooled (two independent samples/pool) before labeling. We also compared the miRNA expression profiles of hMSCs transduced with the lentiviral vector pLV-EmGFP-MIRN335 with MSCs transduced with the control vector pLV-EmGFP-Mock. For the gene expression microarrays, a total of three independent samples were used for each condition.

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Transcriptional effects of LMP3 in MSC

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