Transcriptome analysis and characterization of differential pathways of hMSCs in response to BiFP containing integrin or DDR2 binding motifs creates signals

To demonstrate the capacity of BiFP with specific integrin or receptor binding motifs for signal creation, three individual hMSCs from distinct sources, such as bone marrow (BM-hMSCs) and dental pulp (DP-hMSCs), were cultured in complete growth medium with BiFP containing integrin α2β1 (GFOGER) or discoidin domain receptors 2 (DDR2, GVMGFO) binding motifs for 48 h, followed by RNA-seq analysis. Gene Ontology (GO) and Gene Set Enrichment Analysis (GSEA) revealed that BiFP containing α2β1 or DDR2 binding motifs enriched different gene sets compared to the same cells treated with peptides not containing the binding motifs (Fig. S6A). These data suggest that BiFP containing integrin or receptor binding motifs creates signals that may play a role in instructing cell fate determination. Three individual hMSCs from distinct sources, such as bone marrow (BM-hMSCs) and dental pulp (DP-hMSCs), were cultured in complete growth medium with 40 uM BiFP containing integrin α2β1 (GFOGER) or discoidin domain receptors 2 (DDR2, GVMGFO) binding motifs for 48 h, followed by RNA-seq analysis. (A) Venn diagram showing overlap (pathway numbers) among gene sets associated with three individual hMSCs. (B) Heat map of top 10 significantly shared signaling pathways and bio-functions by gene set enrichment analysis (GSEA) of RNA-seq data associated with integrin or DDR2 binding motifs creates signals that may play a role in instructing cell fate determination. (C) PI3K/AKT activation pathway in the top 10 significantly reactome pathways from overlap integrin-response genes of DP-hMSCs.