Metformin enhances gingival metabolic response in early disease development at single cell level
PD
DISEASE, OXIDATIVE STRESS, PATHOGENESIS
To provide an in-depth analysis of the effect of Metformin preventive effect on gingival cell dynamics when transitioning from health to disease, we transcriptionally profiled single cells derived from mice that underwent Metformin treatment in health and PD induction. We obtained freshly dissected mouse gingival tissue and isolated live cells to be sequenced on the 10 × Genomics Chromium platform for single-cell RNA-seq (scRNA-seq) (Additional file To better visualise the tissue-specific effects of Metformin in health and disease, re-clustering analysis of epithelial and stromal cells was performed to identify the effects of Metformin on sub-clusters transcriptional signature (Fig. scRNAseq analysis of gingival epithelial and stromal compartments during Metformin treatment. Further analysis of the dataset according to the GO related genes revealed significant differences in gene expression in the epithelium and stromal compartments. We first analysed the epithelial and stromal compartments of the animals that did not undergo disease induction. In the epithelium, when compared to the water control, Metformin significantly upregulated Calml3 (a gene important for inhibition of cell apoptosis during oxidative stress [Following that, we investigated the effect of preventive Metformin on the epithelial, stromal and immune cell compartments of animals that underwent PD induction. We first investigated the differences between diseased epithelium and stromal tissues against healthy control epithelial and stromal tissues. Our results showed that in the diseased animals, those cell compartments suffered metabolic changes with significant downregulation of genes associated with the control of ROS damage and glycolysis (Prdx6, Gstm1, Cbr2, Ldhb), mitochondrial respiration and energy (mt-Nd3, mt-Nd2), and cell structural genes (Krt4, Tuba1a) (Additional file Further, when comparing the group that underwent preventive Metformin administration versus the group on water prior to disease induction, our results showed that in the Metformin group, the epithelial cells upregulated Krt76 and Krt6a (genes important for epithelial immune reaction and differentiation [Overall, we demonstrated in silico that Metformin was capable of enhancing the diseased epithelial and stromal cell compartment metabolism towards homeostasis. Moreover, Metformin primed the gingiva with a protective capacity to oxidative damage and via increasing cell energy. Collectively, these observations suggest that Metformin treatment acts as a preventive modulatory drug against PD establishment at the molecular level of the disease pathogenesis.