Background: Sodium glucose co-transporter 2 inhibitors (SGLT2i) are approved anti-diabetic drugs with wide clinical use. In line, a beneficial effect of the SGLT2i Dapa- and Empagliflozin in the cardiovascular (CV) environment has been reported in several clinical trials highlighting a reduction in rates of heart failure (HF) worsening and lower occurrence of death due to CV events in patients suffering from HF with reduced ejection fraction. Contrarily, Ertugliflozin did not trigger cardio-protective results in patients with type 2 diabetes and atherosclerotic CV disease. This also suggests an SGLT2 independent modus operandi of some gliflozins in CV tissue that, however, still remains undisclosed and needs to be defined.

Methods and Results: Initially, using bioinformatic network analysis we screened for deregulated nodes in mass-spectrometry-based proteomics datasets from human endothelial cells treated with Dapagliflozin or Empagliflozin. We discovered a regulation of clusters involved in reactive oxygen homeostasis as well as mitochondrial respiratory chain components. Subsequently, we validated the suggested modes of action using various in vitro as well as ex vivo functional assays and transcriptome evaluations. Reactive oxygen species levels were reduced in primary human umbilical vein endothelial cells (HUVEC) after treatment with Dapagliflozin and Empagliflozin. Decreased basal and maximal respiratory capacity of mitochondria confirmed the interference of Dapagliflozin with the respiratory chain complex. Moreover, Dapagliflozin inhibited migration of HUVEC in wound healing assays but interestingly had no effect on their tube formation capacity in vitro reflecting angiogenic signaling. We validated anti-inflammatory potential of Dapagliflozin using a nuclear factor kB (NFkB)-dependent in vitro reporter assay. Achieving higher translational potential, repression of interleukin-6 transcription in ex vivo cultivated living rat myocardial tissue underlined the impact of Dapagliflozin on inflammatory pathways.

Conclusion: In summary, we screened for the impact of SGLT2i on human endothelial cells applying bioinformatic processing of proteomics data coupled with various in vitro and ex vivo assays. Our findings indicate a modulation of mitochondrial respiration as well as anti-oxidative potential triggered by SGLT2i in endothelial cells. Direct targeting of the NFkB signaling axis was suggested not only in cell culture but also in living myocardial tissue. The results of this study support a direct non-renal mediated cardiovascular effect of selected SGLT2i on the vasculature and grant further insights into potential molecular modes of action of these drugs to support clinical decision making.