Multi-omics analysis reveals attenuation of cellular stress by empagliflozin in high glucose-treated human cardiomyocytes

Abstract

Background: Sodium-glucose cotransporter 2 (SGLT2) inhibitors constitute the gold standard treatment for type 2 diabetes mellitus (T2DM). Among them, empagliflozin (EMPA) has shown beneficial effects against heart failure. Because cardiovascular diseases (mainly diabetic cardiomyopathy) are the leading cause of death in diabetic patients, the use of EMPA could be, simultaneously, cardioprotective and antidiabetic, reducing the risk of death from cardiovascular causes and decreasing the risk of hospitalization for heart failure in T2DM patients. Interestingly, recent studies have shown that EMPA has positive benefits for people with and without diabetes. This finding broadens the scope of EMPA function beyond glucose regulation alone to include a more intricate metabolic process that is, in part, still unknown. Similarly, this significantly increases the number of people with heart diseases who may be eligible for EMPA treatment.

Methods: This study aimed to clarify the metabolic effect of EMPA on the human myocardial cell model by using orthogonal metabolomics, lipidomics, and proteomics approaches. The untargeted and multivariate analysis mimicked the fasting blood sugar level of T2DM patients (hyperglycemia: HG) and in the average blood sugar range (normal glucose: NG), with and without the addition of EMPA.

Results: Results highlighted that EMPA was able to modulate and partially restore the levels of multiple metabolites associated with cellular stress, which were dysregulated in the HG conditions, such as nicotinamide mononucleotide, glucose-6-phosphate, lactic acid, FA 22:6 as well as nucleotide sugars and purine/pyrimidines. Additionally, EMPA regulated the levels of several lipid sub-classes, in particular dihydroceramide and triacylglycerols, which tend to accumulate in HG conditions resulting in lipotoxicity. Finally, EMPA counteracted the dysregulation of endoplasmic reticulum-derived proteins involved in cellular stress management.

Conclusions: These results could suggest an effect of EMPA on different metabolic routes, tending to rescue cardiomyocyte metabolic status towards a healthy phenotype.

Keywords: High glucose; Human cardiomyocytes; Metabolomics; SGLT2i; Type-2-diabetes mellitus.

Fig. 1

PCA scores plots of Metabolomics, Lipidomics, and Proteomics. PCA scores plots have a confidence ellipsis (95%) for each class. The datasets were pre-processed independently for the three modalities: (1) normalized (2) the missing values and zeros were replaced with one-fifth of the minimum value recorded in the data set for that molecule, (3) logarithm values of the base of 10, and (4) autoscaling

Fig. 2

SUMPCA of the three omics datasets

Fig. 3

Graphical representation of confusion matrices obtained from PLS-DA model of the fused modality. The reported disorder matrices refer to PLS-DA performance in training and test phases

Fig. 4

Dot-plots of nicotinamide mononucleotide, Glucose-6-phosphate, Lactic acid, FA 22:6. A–D The reported values refer to normalized data

Fig. 5

Dot-plots of xanthine, adenine, guanine, uridine, thymine, xanthosine, uridine 5′—monophosphate, UDP-D-glucuronic acid, and guanosine 5′—diphosphate-D-mannose. A–I The reported values refer to normalized data

Fig. 6

Dot-plots of Cer 34:0;O2, Cer 40:0;O2, TG 50:1, TG 56:4, TG 58:6, TG 58:8, TG 60:6, TG 60:7, TG 62:4, TG 62:7, TG 62:8, TG 64:7. A-N: The reported values refer to normalized data

Fig. 7

KEGG enrichment. The scatterplot shows the altered pathways of metabolites and lipids following EMPA treatment

Fig. 8

Ontology analysis. A–G Graphical representation of P62820 (Ras-related protein Rab-1A [OS = Homo sapiens]), P04004 (Vitronectin [OS = Homo sapiens]), Q92538 (Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 [OS = Homo sapiens]), Q96N67 (Dedicator of cytokinesis protein 7 [OS = Homo sapiens]), P39687 (Acidic leucine-rich nuclear phosphoprotein 32 family member A [OS = Homo sapiens]), P39748 (Flap endonuclease 1 [OS = Homo sapiens]). The reported values refer to normalized data. In the String pathway of VIP proteins, the blue nodes represent  ER derived proteins