Systemic proteomic and metabolomic signature of the SGLT2 inhibitor dapagliflozin

Abstract

BackgroundDapagliflozin is a member of a new class of medication called gliflozins indicated for the treatment of diabetes. Gliflozins lower blood glucose levels by inhibition of the sodium glucose co‐transporter 2 (SGLT2), which is predominantly expressed in the S1 segment of renal proximal tubules and responsible for >90% of renal glucose reabsorption. Gliflozins also protect the kidney and heart, which cannot be solely explained by blood glucose lowering. To gain further mechanistic insights, we determined the metabolomic and proteomic signature of dapagliflozin in a murine model of diabetes.MethodsMale C57BL/6 non‐diabetic wildtype and littermate Ins2 (Akita) diabetic mice, both fed a Western diet, were given dapagliflozin (10 mg/kg diet) or vehicle for one week (4 groups, N=8/group). Blood and urine glucose levels confirmed the expected treatment response. We analyzed metabolome, proteome and phosphoproteome for a range of tissues (kidney, liver, heart, smooth muscle, white adipose tissue) and body fluids (serum, urine, erythrocytes), and extended the analysis to the gut metaproteome. For untargeted metabolomics, analytes were measured by HILIC‐MS/MS and RP‐MS/MS. For proteomics and phosphoproteomics, TMT labelled (TMT16plex) peptides were separated by high pH reverse phase fractionation and measured by LC‐MS/MS including FAIMS (Thermo Scientific Orbitrap Exploris 480 mass spectrometer).ResultsMulti‐layered omics data were obtained comprising 11341 proteins, 14762 phosphorylation sites, and 217 metabolites. In general, dapagliflozin‐induced changes were more distinct in Western diet‐fed non‐diabetic than diabetic mice. Metabolomics revealed effects of dapagliflozin on amino acid, carbohydrate and lipid metabolism with most changes observed in urine, serum and liver. In comparison, dapagliflozin‐induced perturbations on proteome and phosphoproteome level were observed predominantly in kidney and affected ATP/lipid/carbohydrate/nitrogen metabolism, mitochondria, mitophagy, methyl transferases, methyl CpG‐binding, kinases, deacetylases, endosomes, cell junctions and gene expression. Interestingly, also the gut metaproteome was affected by dapagliflozin with significant changes in Bacteroides and Bifidobacterium species.ConclusionThe SGLT2 inhibitor dapagliflozin induces distinct effects on kidney proteomics, associated with robust metabolomic signatures in plasma, urine and liver, as well as effects on the gut metaproteome. These effects occurred independent of blood glucose levels and support a concept of pleiotropic effects of SGLT2 inhibition that may originate in the kidney but induce wide‐ranged extrarenal metabolic effects and consequences.