Transcriptome analysis reveals synergistic modulation of E-cadherin/N-cadherin in hMSC aggregates chondrogenesis

N-cadherin-mediated cell adhesion is a vital inductor for mesenchymal condensation in chondrogenesis. Recent studies have revealed the involvement of E-cadherin in enhancing the multipotency of mesenchymal stem cells (MSCs) and limb development; however, the signaling crosstalk of E/N-cadherin remains unclear.

This study aimed to explore the synergistic modulation of E/N-cadherin in the chondrogenic differentiation of MSC aggregates.

Human E/N-cadherin-functionalized (hE/N-cad-Fc) poly (lactic-co-glycolic acid) (PLGA) microparticles (hE/N-cad-PLGA) were incorporated into the human MSC (hMSC) aggregates to upregulate the expression of the corresponding endogenous cadherin. The chondrogenic differentiation of the hMSC aggregates was initiated by hE/N-cad-PLGA, controlling the release of transforming growth factor-β (TGF-β). A transcriptome analysis was used to assess differentially expressed genes (DEGs) modulated by hE/N-cad-Fc in hMSC aggregate chondrogenesis. Gene functions and signaling pathways were assessed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The associated biological pathways were assessed by a protein-protein interaction (PPI) network analysis, and the results were further confirmed by real-time quantitative PCR (qPCR) and western blotting.

A total of 1083 DEGs, comprising 111 upregulated and 972 downregulated genes, were discovered to be related to the enhanced chondrogenic differentiation modulated by hE/N-cad-Fc. The GO and KEGG functional enrichment analyses revealed that hE/N-cad-Fc synergistically regulated the p53-related survival signaling pathway. PPI analysis revealed that mitogen-activated protein kinases (MAPK) caspase regulation is a core aspect of the chondrogenic differentiation process, confirmed by western blotting.

To the best of our knowledge, our study is the first to reveal that the synergistic modulation of E/N-cadherin enhances the chondrogenic differentiation of hMSCs via the ERK1/2-p53 signaling axis.