Impact of lncRNA MIR31HG silencing on osteoblastic phenotype modulation: epigenetic regulation of periodontal ligament mesenchymal cells

• Grant number: 22/13949-8
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2023
• End date: February 28, 2026
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Principal Investigator: Denise Carleto Andia
• Grantee: Taís Browne de Miranda
• Host Institution: Vice-Reitoria de Pesquisa e Pós-Graduação. Universidade Paulista (UNIP). São Paulo , SP, Brazil

Abstract

Periodontal ligament-derived cells (PDLCs) are characteristic of mesenchymal cells and, according to the literature, there is heterogeneity in the ability to form extracellular mineralized matrix. However, published studies by our research group indicate that this heterogeneity is related to distinct epigenetic and transcriptional profiles, which can predetermine a given cell phenotype. Therefore, epigenetic and transcriptional profiles are strongly related to the acquisition of a specific osteoblastic phenotype. Long non-coding RNA (lncRNA) participate in epigenetic regulation by binding to chromatin regulatory proteins and may be involved in the modulation of osteogenic genes. The lncRNA MIR31HG was chosen as the target of this project, based on bioinformatics analyzes of epigenomic data generated in the last Grant (FAPESP/University of Birmingham, UK 2017/07944-5) and our hypothesis is that it is associated with reduced training capacity of mineral matrix. Thus, the role of lncRNA MIR31HG will be investigated in epigenetic regulation in PDLCs with low capacity to produce mineral matrix, immortalized and edited by Clustered Regularly Interspaced Short Palindromic Repeats/associated nuclease Cas9 (CRISPR-Cas9). The effect of lncRNA MIR31HG silencing will be investigated on in vitro mineral matrix formation and on the relationship with the Polycomb 2 repressive complex (PRC2), on the epigenetic marker H3K27me3 and on osteogenic markers, by chromatin immunoprecipitation, followed by sequencing (ChIP-seq) and protein levels (WB or MagPix). Epigenetic mechanisms are plastic and reversible and may be promising alternatives for cell- or biomolecule-based therapies, functionalizing implant surfaces or biomaterials, for the treatment of various bone defects. (AU)