The osteogenic capacity and lncRNA/mRNA expression profile of periosteal-derived mesenchymal stem cells of congenital pseudarthrosis of the tibia

Abstract Congenital pseudarthrosis of the tibia (CPT) is an uncommon disease that is difficult to treat. The biological properties, molecular expression patterns and regulatory mechanisms of periosteal-derived mesenchymal stromal cells (PDMSCs) remain unclear. In this study, we separated PDMSCs from CPT patients or control donors. Cell viability and EdU positive proportion in CPT PDMSCs were lower than control. After osteogenic differentiation induction, the ALP staining intensity, calcium nodule formation level and Osterix, Runx2, OCN, OPN protein level were lower in CPT PDMSCs. Co-culture THP1-derived macrophage and PDMSCs, after osteoclast differentiation induction, TRAP level, dissolved hydroxyapatite plate activity, CTSK, CALCR and ITGβ3 levels of co-culture with CPT PDMSCs were shown to be higher than control. RNA-seq results revealed 822 differentially expressed mRNAs and 194 differentially expressed lncRNAs, respectively. Functional analysis showed that differentially expressed lncRNAs involved in tryptophan metabolism, steroid biosynthesis, sphingolipid metabolism and so on. Based on differential mRNAs and lncRNAs, we conducted correlation analysis of their cis regulation, and the results showed that 27 mRNAs were cis regulated with 24 lncRNAs, a total of 30 cis-regulated combinations, including MIR22HG-WDR81, AC103702-HOXB4, AC004080-HoxA13, lncRNA ATG12-CDO1 and lncRNA FAM227B-FGF7. Our results indicat that CPT PDMSCs has weak proliferation and osteogenesis ability, but stronger ability to promotes osteoclast differentiation, and certain lncRNAs and mRNAs were differentially expressed in CPT PDMSCs which have important roles in regulated osteogenesis and osteoclast differentiation.