The Role of miR-342 in Vascular Health. Study in Subclinical Cardiovascular Disease in Mononuclear Cells, Plasma, Inflammatory Cytokines and PANX2

Figure 1. Comparison of data between patients with type 1 diabetes and healthy controls. (a) Levels of IL-7 in plasma, (b) levels of IL-8 in plasma, (c) levels of TNF-α in plasma, (d) levels of VEGF-C in plasma (e) mRNA expression of CXCR1, CXCR2 and PANX2 in PBMCs (f) miR-342-3p and miR-342-5p expression in PBMCs. Data are presented as mean ± SE, analyzed by unpaired t-tests or Mann-Whitney U test accordingly. FC—fold change; HC—healthy control; T1DM—type 1 diabetes mellitus. * p < 0.05, ** p< 0.01 *** p< 0.001.

Figure 2. Correlation between miR-342-3p/-5p and inflammatory markers in type 1 diabetes and healthy controls combined. (a) miR-342-3p and IL-6; (b) miR-342-5p and IL-6; (c) miR-342-3p and IL-8; (d) miR-342-5p and IL-8; (e) miR-342-3p and TNF-α; (f) miR-342-3p and chemokine receptor type 2 (CXCR2). FC—fold change.

Figure 3. Correlation between miR-342-3p/-5p expression in PBMCs and vascular health in type 1 diabetes and healthy controls combined. (a) miR-342-3p and log FAA; (b) miR-342-5p and log FAA; (c) miR-342-3p and CD45dimCD34+CD133+; (d) miR-342-5p and CD45dimCD34+CD133+; (e) miR-342-3p and log TIMP1 (f) miR-342-5p and log TIMP1. FAA—fibronectin adhesion assay; ns—nonsignificant; TIMP-1—tissue inhibitor of metalloproteinases 1.

Figure 4. Correlation between miR-342-3p/-5p and glycemic control in type 1 diabetes and healthy controls combined. (a) miR-342-3p and log HbA1c; (b) miR-342-5p and log HbA1c.

Figure 5. (a) ROC curve for miR-342-3p in type 1 diabetes and healthy controls; (b) ROC curve of HbA1c indicating downregulated miR-342-3p expression; (c) ROC curve for miR-342-5p in type 1 diabetes and healthy controls; (d) ROC curve of HbA1c indicating downregulated miR-342-5p expression.

Figure 6. Ingenuity Pathway Analysis (IPA) prediction network of miR-342-3p/-5p and cardiovascular disease. Downregulation is shown in green and upregulation in red. Orange signifies predicted activation of a molecule or biological function. Paler orange signifies a prediction of less activation. Blue signifies the inhibition of a molecule or biological function. Orange lines represent stimulation, and blue lines inhibition and grey lines reciprocal stimulation and inhibition. Solid uninterrupted lines represent a direct action and dashed interrupted lines represent an indirect action. It has been predicted, through animal knockout studies, that miR-342-5p will activate surfactant protein A1 (SFTPA1) and platelet derived growth factor subunit B (PDGFB) which, in turn, will inhibit heart failure and inflammation. miR-342-3p activates inhibitor of nuclear factor kappa B kinase regulatory subunit gamma (IKBKG), mitogen activated 3 kinase 1 (MAP3K1) and also PDGFB, which are responsible for inhibiting heart failure and inflammation. It is predicted that D-glucose will activate cardiomyopathy, TNF-α and IL-8, and that IL-8 will activate inflammation whilst TNF-α activates VEGF-C as well as…

Figure 7. MiR-342-3p and inflammation. IPA prediction network representing the activation of PANX2 by miR-342-3p and its effect on inflammatory response. Orange signifies the predicted activation of a molecule or biological function. Paler orange signifies a prediction of less activation. Blue signifies the inhibition of a molecule or biological function. Orange lines represent stimulation and blue lines inhibition. Solid uninterrupted lines represent a direct action and dashed interrupted lines represent an indirect action. It was predicted that miR-342-3p would inhibit PANX2. However, when PANX2 was activated, it activated ATP, which was predicted to be responsible for indirectly activating calcium (Ca2+) release to inhibit inflammation. It was also predicted that ATP itself would indirectly inhibit inflammation, and that TNF-α would activate the process of atherosclerosis indirectly, and also activate IL-8, which, in turn, activates inflammation.