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Dental pulp stem cells (DPSCs) are a type of mesenchymal stem cells (MSCs) that are particularly good at making hard tissue, which is needed for teeth. In addition to their dental applications, DPSCs can also differentiate into osteoblasts and form bone, making them useful for engineering bone tissue. Like other MSCs, DPSCs have potential applications in regenerative medicine and tissue engineering.
Prior studies have shown that epigenetic modifications can control gene expression and subsequently guide stem cell differentiation. N6-methyladenosine (m6A)is an epigenetic modification that regulates almost every step of RNA metabolism by adding methyl groups to RNA. This modification is controlled by the methyltransferase complex and plays a critical role in stem cell differentiation and development. Any disruption to m6A deposition can impact stem cell fate and self-renewal.
To better understand how m6A participates in the differentiation of DPSCs, researchers at the Southern Medical University Hospital of Stomatology in China conducted a study to uncover how dynamic m6A marks regulate the fate transition of these cells. Their findings were published in the Journal of Translational Medicine.
In a previous study, the researchers successfully identified the m6A-tagged landscape in immature DPSCs, which they found to be linked to cell senescence and apoptosis. However, they wanted to know how m6A was involved in the differentiation of DPSCs.
They stated, “Clarifying the RNA epigenetic mechanism during DPSC differentiation and manipulating the key modulators in therapeutic applications would advance vital pulp therapy.”
Using DPSCs collected from extracted teeth, the cells were then induced for osteo/odontogenic differentiation. Total m6A was then measured from the extracted RNA using EpigenTek’s m6A RNA Methylation kit, followed by LC-MS/MS analysis to assess the metabolite compounds related to m6A methylation.
The isolated RNA from the DPSCs was also subjected to m6A RIP-seq and m6A RIP-qPCR to investigate the occurrence of m6A modification during mineralization and to examine the role of METTL3, a key enzyme involved in m6A modification. In addition, RNA-sequencing was performed using the Illumina deep sequence platform to obtain RNA profiles.
The data from the study showed that total m6A and epitranscriptomic profile of m6A-tagged mRNA were altered during DPSC differentiation, with increased levels of total m6A in the RNA pool after osteo/odontogenic induction.
In addition, m6A RIP-seq analysis revealed changes in DNA regions and molecule expression related to signal transduction, transcriptional regulation, and cell differentiation. And METTL3 was found to be upregulated during DPSC mineralization, impacting odontogenic differentiation and tissue formation.
In particular, METTL3 was found to modify a specific RNA transcript encoding the protein NOG, which is involved in tooth development, and promotes its degradation by shortening its poly(A) tail, a part of the RNA molecule. A knockdown of METTL3 showed altered NOG expression and downstream signaling pathway changes.
Overall, the study’s investigation revealed that m6A RNA methylation, carried out by the METTL3 enzyme, significantly impacts how DPSCs differentiate into specialized cells, like tooth and bone tissue. This modification alters the activity, stability, and length of RNA molecules, providing a new understanding of dental stem cell behavior and gene regulation during tooth development.
Reference:
Luo H et. al.(December 2022). Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation.J Transl Med.20(1):605.
