Kearney Michaela, Duncan Henry F
Division of Restorative Dentistry and Periodontology, Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland.
Methods Mol Biol. 2023;2588:353-367. doi: 10.1007/978-1-0716-2780-8_20.
Recently, the central role of microRNAs (miRNAs) and epigenetic modifiers in biological and pathological processes, such as stem cell differentiation and inflammation, has stimulated interest. In particular, their influence in dental pulp stem cell (DPSC) differentiation has been highlighted as an exciting avenue of research in the field of Regenerative Endodontics. Although specific miRNAs have been shown to be altered in expression during dental pulp mineralization and repair processes, their interaction with epigenetic modifiers, such as histone deacetylase inhibitors (HDACi) or DNA methyltransferase inhibitors (DNMTi), has not been explored. Currently available next-generation sequencing (NGS) technologies offer the potential to explore in detail the miRNA expression profile of cells, and to investigate the effects of pharmacological inhibitors such as epigenetic modifiers on this profile. This chapter describes the experimental methods required to induce mineralization of DPCs in the presence and absence of epigenetic modifiers and analyze the resulting miRNA expression profiles using RNA sequencing (RNAseq), with a focus on bioinformatic analysis.
最近，微小RNA（miRNA）和表观遗传修饰因子在生物和病理过程（如干细胞分化和炎症）中的核心作用引起了人们的兴趣。特别是，它们在牙髓干细胞（DPSC）分化中的影响已被视为再生牙髓病学领域一个令人兴奋的研究方向。尽管已表明特定的miRNA在牙髓矿化和修复过程中表达会发生改变，但其与表观遗传修饰因子（如组蛋白去乙酰化酶抑制剂（HDACi）或DNA甲基转移酶抑制剂（DNMTi））的相互作用尚未得到研究。目前可用的下一代测序（NGS）技术为详细探索细胞的miRNA表达谱以及研究诸如表观遗传修饰因子等药理抑制剂对该谱的影响提供了可能。本章介绍了在存在和不存在表观遗传修饰因子的情况下诱导牙髓细胞矿化并使用RNA测序（RNAseq）分析所得miRNA表达谱所需的实验方法，重点是生物信息学分析。
