A Quantitative Proteomic Approach for Elucidating the Effects of Antidiabetic Drugs on Coronary Artery Endothelial Cells

Individuals with type 2 Diabetes Mellitus (DMT2) possess an elevated risk of mortality due to cardiovascular disease (CVD). Although there is available a repertoire of antidiabetic drugs (ADD) to treat the disease, mortality risk reduction remains an elusive goal despite improvement in glucose metabolism by drug therapy. Moreover, some drugs further increase the CVD risk in subjects with T2DM, while others appear to partially reduce the risk. Hence, new approaches and thinking are needed to explore this phenomenon beyond the classical drug’s antihyperglycemic effects and principal organs involved in glucose metabolism. In this study, we explored the effects of selected ADD (with epidemiological CVD evidence in the clinic) on Human Coronary Artery Endothelial Cells (HCAEC) —involved in the development of CVD— using a quantitative SWATH‐based proteomic and comprehensive bioinformatic approach. To create a template or network of pathways linked to cell damage in HCAEC, we generated a list of proteins modulated (Up and down‐regulated) by the treatment with the pro‐inflammatory cytokine TNF‐a and performed a Functional Enrichment Analysis against REACTOME and KEEG Databases using ClueGO software. We further expanded our network by utilizing reported data of endothelial cells treated with cytokine IL‐1B. ADD treatment consisted of cells incubated with physiological concentrations of either metformin (100 μM), rosiglitazone (1 μM), empagliflozin (1 μM), saxagliptin (1 μM), and vehicle (DMSO) for 48 h. The list of proteins modulated by the ADD was analyzed by ClueGO using as a template the pathway network of cytokines. Our results hint at anti‐inflammatory actions of all ADD tested, but at different levels within the immune system network. Rosiglitazone, followed by saxagliptin, modulated more proteins linked to anti‐inflammatory actions compared to the rest of the drugs. Neutrophil degranulation, signaling by interleukins, and interferon signaling were modulated by rosiglitazone and saxagliptin, while the rest of the drugs modulated less or none of these specific pathways. Also, ADD down‐modulated pathways associated with the metabolism of proteins and angiogenesis. In conclusion, our study provides insight into the pathways on HCAEC modulated by drugs used to treat subjects with T2DM. Further, our results contribute to the understanding of molecular mechanisms, that could explain the clinical effects noted with some ADD for which there is no clear mechanistic data beyond their classical antihyperglycemic effects. Support or Funding Information Research funded by an Internal Project to AMU and a Graduate Scholarship from the Mexican National Council for Science and Technology (CONACyT) to CMM