Aberrantly expressed long noncoding RNAs in adipose-derived mesenchymal stem cells differentiation to nucleus pulposus-like cells.

Stem cell-based therapy holds promise for intervertebral disc degeneration (IDD) regeneration, but the underlying mechanisms require further elucidation. Long noncoding RNAs (lncRNAs) have been implicated in physiological processes such as apoptosis and differentiation. While numerous studies have investigated messenger RNAs (mRNAs) and lncRNAs in normal versus degenerated nucleus pulposus (NP), the differential expression of lncRNAs and mRNAs during stem cell differentiation into NP-like cells remains poorly characterized. This study aimed to identify differentially expressed mRNAs and lncRNAs during the differentiation of human adipose-derived mesenchymal stem cells (hADSCs) into NP-like cells and to explore the associated signaling pathways and regulatory networks. hADSCs were induced to differentiate into NP-like cells using a cytokine cocktail. Differentiation was confirmed by assessing marker gene expression via PCR and immunofluorescence staining. RNA sequencing (RNA-seq) was employed to profile lncRNA and mRNA expression during differentiation. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were used to identify the biological functions and pathways associated with the significantly altered genes. 500 lncRNAs (217 up and 283 down) and 601 mRNAs (269 up and 332 down) were significantly differentially expressed during hADSC differentiation. The RNA-seq findings were validated by real-time PCR. GO analysis revealed significant enrichment of terms related to biological processes, molecular functions, and cellular components. KEGG pathway analysis highlighted several significantly enriched pathways. A gene interaction network illustrated relationships among the differentially expressed genes, and an mRNA-lncRNA co-expression network suggested potential regulatory interactions. This study is the first to profile differentially expressed lncRNAs and mRNAs during hADSC differentiation into NP-like cells. These findings provide valuable insights for understanding the mechanisms of stem cell therapy and IDD regeneration.