scRNAseq characterization of synovial stromal cells in mouse STIA
Objective: Upregulation of glycolysis and glutaminolysis in the rheumatoid arthritis fibroblast-like synoviocytes (RAFLS) has already been established by existing literature . Henceforth, this study was undertaken to determine if dual inhibition of these processes by novel phytobiologic c28MS have the potential of promising  synergistic therapy for rheumatoid arthritis by targeting the underlying metabolic reprogramming as existing treatment strategies for RA do not target dual inhibition. Methods: Gene associations with HK2 and GLS expression were identified by examining murine and human available scRNA-seq. The metabolic profile of FLS cells was determined by 1H magnetic resonance spectroscopy under conditions of glycolytic and glutaminolysis inhibition and HK2 and GLS1 expression was evaluated by western blot. FLS functional analysis was conducted under similar inhibitory conditions. ELISA was employed for the quantification of pivotal markers namely IL6, CCL2 and MMP3. For arthritis experiments, mice were injected with K/BxN sera on Day 0 and treated for 10 days. Results: scRNAseq data identified... Metabolomic and functional analysis revealed that c28MS had the potential of checking the aggressive behavior of RAFLS by targeting upregulated glycolysis and glutaminolysis thereby inhibiting them. Target expression remained unaffected by the action of the dual inhibitor suggesting only potential blockage. Furthermore, the compound was able to inhibit inflammatory arthritis by decreasing the clinical score and paw inflammation in the K/BxN model. Conclusion: Our findings imply that dual inhibition of glycolysis and glutaminolysis could be an effective strategy in the treatment of RA with chances of greater remission. It also suggests that targeting more than one pathway in the metabolome can be a novel treatment approach in non-cancer scenarios. Overall design: STIA was induced by intravenous injection of 100 µl arthritogenic serum from KRN mice (K/BxN). Cells were extracted at various timepoints (rest d0, peak d7-9, resolving d15, resolved d22) followed by CD45neg purification of stromal cells and 3' mRNA scRNAseq (10x Genomics)