"Epigenetic regulation of osteogenic potential in mesenchymal stem cells derived from periodontal ligament: ATAC-Seq and Bioinformatic analysis"

Abstract

Since human multipotent periodontal ligament stem cells (hPDLSCs) can give rise to bone, they can be promising candidates for regenerative therapies in a broad range of bone diseases. However, hPDLSCs can show distinct osteogenic potential, which might be a clinical issue. The molecular mechanisms underlying their osteogenesis are not well understood, with limited information regarding the epigenetic background and regulation. Since cell fate is determined by lineage specific gene regulation and underlying epigenetic mechanisms such information is indispensable to safely apply hPDLSCs in human clinical trials. The chromatin regions with accessible chromatin will be identified in hPDLSCs presenting high and low osteogenic potential, after 3 and 10 days of osteogenesis induction, by the Assay for Transposase Accessible Chromatin (ATAC), followed by next generation sequencing (Seq) and Bioinformatic analysis. These chromatin regions are indicative of active regulatory elements and transcription factor binding sites which are related to changes in the gene expression needed to osteogenesis. The study groups will be DMEM (control, non-induced) and OM (osteogenic media, induced). The ATAC-Seq, together with the DNA (hydroxy)methylome proposed in the PhD project and RNA-Seq and ChIP-Seq proposed in the main research project with should fill the knowledge gaps in the fields of PDLSCs basic biology and bone formation/metabolism. We aim to facilitate the clinical application of PDLSCs by proposing markers of high osteogenic potential for further validation.