MicroRNA-221 silencing attenuates the degenerated phenotype of intervertebral disc cells

Abstract

The aim of this study was to investigate the role of an antichondrogenic factor, MIR221 (miR-221), in intervertebral disc degeneration (IDD), and provide basic information for the development of a therapeutic strategy for the disc repair based on specific nucleic acid based drugs, such as miR-221 silencing. We established a relatively quick protocol to minimize artifacts from extended in vitro culture, without selecting the different types of cells from intervertebral disc (IVD) or completely disrupting extracellular matrix (ECM), but by using the whole cell population with a part of resident ECM. During the de-differentiation process miR-221 expression significantly increased. We demonstrated the effectiveness of miR-221 silencing in driving the cells towards chondrogenic lineage. AntagomiR-221 treated cells showed in fact a significant increase of expression of typical chondrogenic markers including COL2A1, ACAN and SOX9, whose loss is associated with IDD. Moreover, antagomiR-221 treatment restored FOXO3 expression and increased TRPS1 expression levels attenuating the severity grade of degeneration, and demonstrating in a context of tissue degeneration and inflammation not investigated before, that FOXO3 is target of miR-221. Data of present study are promising in the definition of new molecules useful as potential intradiscal injectable biological agents.

CircCOG8 Downregulation Contributes to the Compression-Induced Intervertebral Disk Degeneration by Targeting miR-182-5p and FOXO3

“…FOXO3, known as FOXO3a or FKHR-L1, is among the Forkhead box O (FOXO) family of transcription factors which plays a crucial part in the normal physiological structure and function of IVD (Alvarez-Garcia et al, 2018). Previous study revealed that the FOXO3 expression level was obviously downregulated in severe degenerated human IVDs, contributing to the major histopathological changes during IDD progression (Penolazzi et al, 2018). In the present study, we observed that the FOXO3 expression in the NP cells was repressed by compression loading.…”

“…It has been reported that the transcription factors of the Forkhead box O class (FOXOs) played important regulating roles in the intervertebral disk maturation and homeostasis (Alvarez-Garcia et al, 2018;Penolazzi et al, 2018). Using the TargetScan online tool, we found that the Forkhead box O3 (FOXO3) possessed complementary sequence to the miR-182-5p seed region ( Figure 5A).…”

Extracellular Matrix From Decellularized Wharton’s Jelly Improves the Behavior of Cells From Degenerated Intervertebral Disc

“…P0 cells were expanded by growing for a period not exceeding a week until subconfluent, detaching by trypsinization, and maintained in culture for more than two passages to obtain IVD cells that were used for later experiments. As already demonstrated after monolayer culture expansion, cells isolated from human lumbar IVD become de-differentiated, lose their chondrogenic-like phenotype resembling the degeneration process (Penolazzi et al, 2018).…”

Exosomal LncRNA–NEAT1 derived from MIF-treated mesenchymal stem cells protected against doxorubicin-induced cardiac senescence through sponging miR-221-3p

“…Current insights into the molecular systems of lncRNA–miRNA regulatory interactions and implications in the treatment of Dox-induced cardiotoxicity allow a hypothesis that lncRNA–NEAT1 may be involved in the regulatory network of mRNA via competing endogenous RNAs (ceRNAs)-mediated miRNA evasion [ 22 ]. A target ceRNA of lncRNA–NEAT1, miR-221-3p, impaired tissue regeneration through inhibiting the sirtuin (Sirt) family of proteins [ 23 ]. A previous study reported that exosome-transferred LncRNA–NEAT1 activated the Sirt protein family to exert a cardioprotective effect [ 19 ].…”