Comprehensive investigation of (redox-) proteomic changes induced by cana-, dapa-, empagliflozin treatment in cardiac cells

ABSTRACT: One of the major macrovascular complications that can occur in type 2 diabetes is the development of cardiovascular diseases as for instance heart failure. However, sodium-glucose co-transporter-2 (SGLT-2) inhibitors have shortly after their approval shown to decrease death rates in diabetic patients. Later on, this effect was proven to be independent of the diabetes status of a patient. Interestingly, SGLT-2 is not expressed in cardiac tissue and the underlying mechanisms for these effects remain unclear. In our study we treated two cardiac cell lines (AC16 and HCM) with three different SGLT-2 inhibitors (cana-, dapa and empagliflozin) to investigate their influence on the (redox) proteome. Besides concentrations comparable to those reached in patients (100 nM of cana-, and 10 nM of dapa- or empagliflozin), we also applied a higher dose (10 μM) to check if any effects would be concentration dependent. Furthermore, we included stem-cell derived cardioids on which we mimicked reperfusion injury with the aim to investigate whether and how empagliflozin (20 nM) help to cope with a lack of and subsequent reperfusion with oxygen. These data are included in PRIDE project PXD063242.We were able to show that drug treatments led to higher amounts of glutathione and related thiol metabolites (additionally measured, data not included), presumably showing an increased cellular capacity for handling redox-stress. On the proteome level we were mostly interested in finding shared proteins or redox affected cysteine sites across drugs and systems. For quantitative proteomics we found 21 proteins shared between all 10 μM drug treatments in HCM cells and cardioids, of which 16 showed a consistent expression pattern (downregulation). Among them were glutathione reductase (GSR), peroxiredoxin 2 (PRDX2), ragulator complex protein LAMTOR5 and the catalytic subunit of serine/threonine-protein phosphatase 2A (PP2A). On redox proteome level of HCM cells we detected 54 shared cysteine sites affected by redox changes, among them C423/424 in PKM2, C73 in TXN as well as C100 and C204 in PRDX5, which are the core of its peroxidative activity. All of the mentioned sites were more oxidized upon 10 μM drug treatments. In AC16 cells C93 in MDH2, C127 in PPP1CA and both sites of PRDX5 found in HCM cells as well.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Tamara Tomin  

LAB HEAD: Ruth Birner-Gruenberger

PROVIDER: PXD063467 | Pride | 2026-01-20

REPOSITORIES: Pride