Perturbation of interactome through micro-RNA and methylome analysis in diabetes endophenotypes: the PIRAMIDE pathogenic clinical study design

Background:The main challenge in type 2 diabetes (T2D) is to detect the regulators of pathogenic events during early stages of disease, as well as prevention and progression follow-up of cardiovascular (CV) complications. DNA methylation and micro-RNAs (miRNAs) are major components of the epigenome, which are involved in the diabetic interactome. This study protocol may contribute to advance our knowledge on molecular basis underlying T2D and its CV complications, as well as provide putative useful prognostic biomarkers.Methods:The perturbation of interactome through micro-RNA and methylome analysis in diabetes endophenotypes: the PIRAMIDE pathogenic clinical study protocol is a cross-sectional research program planned to combine big data and network-based analysis aimed to investigate whether DNA methylation and miRNAs may act as simultaneous regulators of the interactome in T2D patients. Clinical datasets will be aggregate to large-scale DNA methylation, mRNA-Seq, and miRNA-Seq analysis performed both on purified CD4+and CD8+ T cells isolated from 35 T2D patients and 35 sex and age-matched controls. DNA methylome data will be used as input for the weighted human DNA methylation PPI network (WMPN) algorithm. RNA sequencing data will be used as input data for the TargetScan algorithm. The primary endpoint will be to integrate both DNA methylation and miRNA networks to potentially capture which genes are simultaneously modulate by interactions between epigenetic changes. Then,statistical analysis will be performed to correlate these molecular modifications with development of T2D-related CV complications. Conclusions: PIRAMIDE pathogenic clinical study protocol will test the hypothesis that simultaneous interactions between DNA methylation and miRNAs may hit T2D-associated candidate genes and predict the development of T2D-related CV complications.Trial Registration:The ongoing PIRAMIDE pathogenic clinical study protocol has been registered on NIH website (NCT03792607)

Left ventricular function in the successive phases of systemic hypertension evaluated with pulsed Doppler echocardiography

Systolic and diastolic function is impaired in patients with hypertensive heart disease. Systolic hypertension induces a succession of LV hemodynamic changes and can be regarded as a spectrum from maladaptive hypertophy to heart failure. The left ventricular hemdynamic changes that occur can be measured non-invasively by Doppler echocardiography. The aim of the study was to hemodynamically characterize the different phases of left ventricular [LV] function in patients affected by systemic hypertension [SH]. 95 normotensive healthy controls [group I] and 94 hypertensives [group II] were enrolled. Hypertensive patients were divided in two sub-groups according to echocardiographic signs of left ventricular hypertrophy [LVH]. Other echocardiographic parameters measured using tissue Doppler were Isovolumic Relaxation Time [IRT], isovolumic contraction time [ICT], and systolic motion [Sm]. Myocardial Performance Index [MPI] using Tissue Doppler Echocardiography [TDE] was defined in both the control group and the two hypertensive subgroups. Ejection fraction [EF] was also calculated in all participants. An increased MPI derived from the rise of isovolumetric relaxation time [IRT] was found in hypertensives without LVH [sub-group II-a], whereas isovolumetric contraction time [ICT] and Systolic motion [Sm] were unchanged. Hypertensive patients with LVH demonstrated more prominent increase of MPI, increase in IRT-prolongation, ICT-increase and Sm-decrease. The results obtained indicate impaired relaxation in sub-group II-a. On the contrary, a systolo-diastolic LV dysfunction was found in sub-group II-b. E.F decreased in this same sub-group of hypertensives in comparison with controls and sub-group II-a, as a sign of maladaptive LVH evolving towards heart failure. Doppler echocardiography appears able to distinguish the different forms and degrees of LV dysfunction in SH in relation to the different phases of the hypertensive disease process

Myocardial performance index evaluated with tissue doppler echocardiography at early, intermediate and late phase of acute myocardial infarction

In this study, ejection fraction% [EF%] and myocardial performance index [MPI] were recorded in 67 survivors at early, intermediate and late phase of acute myocardial infarction [AMI] .EF% was echocardiographically obtained by the Simpson's method; MPI was calculated using Tissue Doppler Echocardiography [IDE] derived from isovolumetric contraction time [ICT]; isovolumetric relaxation time [IRT] and ejection time [ET]. Results were compared with those obtained in 70 controls matched for age and sex.At hospital discharge [early evaluation], EF% was < 50% with significant increase in MPI in respect to the healthy controls [increase in ICT, significant reduction in ET and IRT was unchanged]. Six months later [intermediate evaluation], EF% still resulted in < 50%, MPI was slightly reduced with further increase in ICT and IRT in comparison to the early evaluation, and slight reduction in ET Finally, one year later [late evaluation], in spite of increase in EF>50%, MPI was still increased, with slight rise in ICT, almost normalization in ET, but more evident increase in IRT. The outcomes of MPI demonstrate that in post-AMI patients, late prevalent diastolic ventricular dysfunction occurs following an early systolic dysfunction. In this study, EF% appears to be less sensitive than MPI in defining late post-AMI left ventricular dysfunction. Finally,TDE seems to be more sensitive than conventional Doppler method in measuring MPI

Post-necrotic left ventricular dysfunction in Diabetes Mellitus: effects of trimetazidine

The Index of Myocardial Performance [IMP] in 149 non-diabetic [group I] and 151 diabetic [group II] subjects who were treated for acute myocardial infarction was evaluated using two-dimensional Doppler echocardiography. Isovolumetric Contraction Time [ICT], Isovolumetric Relaxation Time [IRT] and Ejection Time [ET] were also measured. All patients in both groups received conventional, anti-ischaemic therapy [nitrates, ACE-inhibitors, and antiplatelet drug]. In addition, 74 patients in group II [subgroup IIa] received an oral dose of 20 mg of trimetazidine, three times daily. The remaining 77 diabetics in group II were treated with conventional drugs alone [subgroup IIb]. All diabetic patients [group II] also received an anti-diabetic [oral drug or insulin] treatment to keep their diabetes under control. Twelve months after the experiment, IMP was significantly [p<0.001] higher in diabetic patients [0.55 +/- 0.05] compared to non-diabetic controls [0.49 +/- 0.04]. IRT was similar in both groups [81 +/- 15 ms vs 83 +/- 12 ms] and ET [275 +/- 27 ms vs 295 +/- 29 ms] was decreased in diabetics compared to the control group. The one-year follow-up showed a significant decrease in IMP in patients treated with trimetazidine [subgroup IIa] compared to those treated with conventional drugs [subgroup IIb alone]. IRT values were lower in sub-group IIa compared to that of subgroup IIb. ICT returned towards the normal limits in both subgroups. Finally, ET decreased in subgroup IIa but increased in subgroup IIb compared to values obtained at the onset of treatment. In conclusion, trimetazidine when added to the conventional, anti-ischaemic therapy, seems to induce a more evident attenuation of post-AMI left ventricular dysfunction compared to those not given the drug