Michał Kacprzak

Proteomic analyses based on mass spectrometry provide a powerful tool for the simultaneous identification of proteins and their signatures. Disorders detection at the molecular level delivers an immense impact for a better understanding of the pathogenesis and etiology of various diseases. Acute coronary syndrome (ACS) refers to a group of heart diseases generally associated with rupture of an atherosclerotic plaque and partial or complete thrombotic obstruction of the blood flow in the infarct-related coronary artery. The essential role in the pathogenesis of ACS is related to the abnormal, pathological activation of blood platelets. The multifactorial and complex character of ACS indicates the need to explain the molecular mechanisms responsible for thrombosis. In our study, we performed screening and comparative analysis of platelet proteome from ACS patients and healthy donors. Two-dimensional fluorescence difference gel electrophoresis and nanoscale liquid chromatography coupled to tandem mass spectrometry showed altered expressions of six proteins (i.e., vinculin, transgelin-2, fibrinogen β and γ chains, apolipoprotein a1, and tubulin β), with the overlapping increased expression at the mRNA level for transgelin-2. Dysregulation in protein expression identified in our study may be associated with an increased risk of thrombotic events, correlated with a higher aggregability of blood platelets and induced shape change, thus explaining the phenomenon of the hyperreactivity of blood platelets in ACS.

The pathophysiology of atherosclerosis and acute coronary syndrome (ACS) is related to interactions between immune cells, endothelium, and blood platelets. An increasing number of reports confirm the link between excessive immune activation and cellular cross-talk with ACS incidence. Our genetic and proteomic analysis was performed on strictly selected atherosclerotic patients with non-fatal ACS without typical risk factors and healthy donors. Results showed changes in the gene expression levels of the various inflammatory factors derived from the peripheral blood cells that drive the over-activation of the immune system. The enhanced activation of the immune system may lead to the overexpression of the pro-inflammatory mediators, which causes self-perpetuating machinery of processes associated with thrombosis. In our preliminary study, we confirmed an altered expression of genes associated with the inflammation and overall interaction of the vascular microenvironment. Furthermore, 5 of 92 analyzed genes, CCL2, CCR2, CSF2, GZMB, and ICOS, were expressed only in patients with ACS. In conclusion, the augmented expression of the pro-inflammatory genes from the peripheral blood cells may be a crucial genetic factor leading to the occurrence of acute inflammation and thus be significant in ACS pathogenesis.

The pathological conditions caused by blood platelet activation constitute a fundamental core in the pathogenesis of Acute Coronary Syndrome (ACS). The hyperactivity of platelets in ACS is well-documented, but there is still little research into the molecular basis of phenotypic changes in platelet functionality. To expand the knowledge of this phenomenon, we analyzed the disturbances in the expression of several key platelet receptors and the aspect of regulating potential abnormalities. Platelet surface receptors are responsible for maintaining the hemostatic balance, platelet interaction with immune cells, and support of the coagulation cascade leading to occlusion of the vessel lumen. Due to their prominent role, platelet receptors constitute a major target in pharmacological treatment. Our work aimed to identify the molecular alteration of platelet surface receptors, which showed augmented mRNA expression of P2Y12, GP1BB, ITGA2B, and ITGB3 and increased protein concentrations of P2Y12 and GP IIb/IIIa in ACS. The upregulation of the P2Y12 level was also confirmed by confocal and cytometric visualization. Furthermore, we evaluated the expression of two microRNAs: miR-223-3p and miR-126-3p, which were suggested to regulate platelet P2Y12 expression. Results of our study present new insight into the molecular background of ACS.

Transcriptome analysis constitutes one of the major methods of elucidation of the genetic basis underlying the pathogenesis of various diseases. The post-transcriptional regulation of gene expression is mainly provided by microRNAs. Their remarkable stability in biological fluids and their high sensitivity to disease alteration indicates their potential role as biomarkers. Given the high mortality and morbidity of cardiovascular diseases, novel predictive biomarkers are sorely needed. Our study focuses for the first time on assessing potential biomarkers of acute coronary syndrome (ACS) based on the microRNA profiles of platelets. The study showed the overexpression of eight platelet microRNAs in ACS (miR-142-3p; miR-107; miR-338-3p, miR-223-3p, miR-21-5p, miR-130b-3p, miR-301a-3p, miR-221-3p) associated with platelet reactivity and functionality. Our results show that the combined model based on miR-142-3p and aspartate transaminase reached 82% sensitivity and 88% specificity in the differentiation of the studied groups. Furthermore, the analyzed miRNAs were shown to cluster into two orthogonal groups, regulated by two different biological factors. Bioinformatic analysis demonstrated that one group of microRNAs may be associated with the physiological processes of platelets, whereas the other group may be linked to platelet–vascular environment interactions. This analysis paves the way towards a better understanding of the role of platelet microRNAs in ACS pathophysiology and better modeling of the risk of ACS.

Atrial natriuretic peptide (ANP) is secreted in response to the stretching of the atrial wall. Atrial ischemia most likely impairs the ability of atrial myocytes to produce ANP. Atrial infarction (AI) is rarely diagnosed but not infrequently associated with myocardial infarction (MI). The aim of the study was to assess the association between AI and the prognostic value of N-terminal proANP (NT-proANP) in patients with MI treated with primary percutaneous coronary intervention (PCI). We evaluated data of 100 consecutive patients. Plasma levels of NT-proANP were measured by the ELISA method. ECG recordings were interpreted to diagnose AI according to Liu’s criteria. All patients were followed-up prospectively for 12 months for the manifestation of major adverse cardiovascular events (MACE), defined as unplanned coronary revascularization, stroke, reinfarction or all-cause death. AI was diagnosed in 36 patients. 14% of patients developed MACE. AI did not affect the incidence of MACE or any of its components, nor the patients’ prognosis. NT-proANP revealed to be a strong predictor of death but was not associated with other adverse events. Conclusions: AI in patients with MI treated with primary PCI is not connected with their prognosis nor affects the usefulness of NT-proANP in predicting death during the 12-month follow-up.

Background: In the era of modern interventional cardiology implantation of balloon expandable stent is a finishing touch of almost every coronary angioplasty. However sometimes we face clinical situation when decision on stent diameter is complicated, especially in ectatic part of arteries, in situations when artery lumen is obscured with thrombus or when reference diameter of proximal and distal part of lesion vary greatly. That is why idea of self-apposing stent similar to one used in peripheral vascular interventions was adopted into cardiology. The aim of this study was to present single center registry of the STENTYS® stent implantation in 40 selected patients with acute coronary syndromes (ACS) or with stable angina (CAD) treated with this self-expandable stent Methods and Results: The device was successfully implanted in all patients. During in hospital observation and 30 day follow-up there were 2 cases of death, none of the patient had acute stent thrombosis or ACS STEMI . In one case ACS type 4b was diagnosed. In all patients stent was delivered in target lesion. In two cases procedure was performed in patients with multivessel coronary artery disease extending into left main stem in state of cardiogenic shock,. These patients died immediately after procedure. There were two procedure complications, in one case dissection after post dilatation occurred distally to the stent and in one patient calcified proximal part of LAD was dissected with system passage. 38 patients survived 12 moth follow-up period, 3 patients (7,8%) underwent repeated target-lesion revascularization Conclusions: In presented single center registry STENTYS® stent was used with a high delivery and procedural success rate. Satisfactory clinical long-term outcome both in stable patients and ACS-patients with the repeated revascularization ratio of 7.8 % was observed. Stent design allowed a successful treatment of bifurcation lesions. Background: In the era of modern interventional cardiology implantation of balloon expandable stent is a finishing touch of almost every coronary angioplasty. However sometimes we face clinical situation when decision on stent diameter is complicated, especially in ectatic part of arteries, in situations when artery lumen is obscured with thrombus or when reference diameter of proximal and distal part of lesion vary greatly. That is why idea of self-apposing stent similar to one used in peripheral vascular interventions was adopted into cardiology. The aim of this study was to present single center registry of the STENTYS® stent implantation in 40 selected patients with acute coronary syndromes (ACS) or with stable angina (CAD) treated with this self-expandable stent Methods and Results: The device was successfully implanted in all patients. During in hospital observation and 30 day follow-up there were 2 cases of death, none of the patient had acute stent thrombosis or ACS STEMI . In one case ACS type 4b was diagnosed. In all patients stent was delivered in target lesion. In two cases procedure was performed in patients with multivessel coronary artery disease extending into left main stem in state of cardiogenic shock,. These patients died immediately after procedure. There were two procedure complications, in one case dissection after post dilatation occurred distally to the stent and in one patient calcified proximal part of LAD was dissected with system passage. 38 patients survived 12 moth follow-up period, 3 patients (7,8%) underwent repeated target-lesion revascularization Conclusions: In presented single center registry STENTYS® stent was used with a high delivery and procedural success rate. Satisfactory clinical long-term outcome both in stable patients and ACS-patients with the repeated revascularization ratio of 7.8 % was observed. Stent design allowed a successful treatment of bifurcation lesions.

Context: Cardiogenic shock (CS) still remains one of the main causes of death in acute myocardial infarction (AMI).Objective: The aim of this study was to evaluate the prognostic value of midregional (MR)-proadrenomedullin in AMI complicated by CS.Methods: Forty-seven consecutive patients were included in our prospective observational study. All patients underwent coronarography and successful percutaneous coronary intervention (PCI). Plasma levels of MR-proADM were measured by a immunofluorescence method. The primary endpoint of the study, defined as cardiovascular death, occurred in 17 patients (36%).Results and conclusion: Elevated plasma level of MR-proADM, determined 24h after diagnosis of CS could be a predictor of in-hospital mortality in patients with AMI complicated by CS.

Background: Elevated mean platelet volume (MPV) has been recently discussed as a predictorof death in patients with acute coronary syndrome (ACS), but the cut-off point of MPV inrelation to poor prognosis has not been estimated so far. The aim of this study was to evaluate MPV and its prognostic value in ACS complicated by cardiogenic shock (CS). Such an analysisin patients with the most serious and fatal complication of ACS has not been performed inpreviously published research. Methods: Fifty three patients with ACS complicated by CS (age 68.9 ± 11.4, 49% women, 92% STEMI, 55% fatal CS) and 53 age- and gender-matched patients with uncomplicated ACS as a control group (age 69.1 ± 10.6, 49% women, 92% STEMI, 0% fatal) were includedin our prospective study from 2010 to 2012. All the patients underwent successful primary percutaneous coronary intervention. MPV was determined on admission (MPV1) and in consecutive two days of hospitalization (MPV2, MPV3). The blood sample was analyzed immediately after collection in EDTA tubes using an automatic blood counter. Results: MPV1 was similar in both groups (8.91 ± 1.11 fl vs. 8.57 ± 0.74 fl, NS). Furthermore, there were no statistically significant differences in MPV value in fatal and nonfatal CS(8.90 ± 1.18 fl vs. 8.93 ± 1.05 fl, NS). Conclusions: The above results suggest that MPV cannot be considered a predictor of poorin-hospital outcome in patients with ACS complicated by cardiogenic shock. Background: Elevated mean platelet volume (MPV) has been recently discussed as a predictorof death in patients with acute coronary syndrome (ACS), but the cut-off point of MPV inrelation to poor prognosis has not been estimated so far. The aim of this study was to evaluate MPV and its prognostic value in ACS complicated by cardiogenic shock (CS). Such an analysisin patients with the most serious and fatal complication of ACS has not been performed inpreviously published research. Methods: Fifty three patients with ACS complicated by CS (age 68.9 ± 11.4, 49% women, 92% STEMI, 55% fatal CS) and 53 age- and gender-matched patients with uncomplicated ACS as a control group (age 69.1 ± 10.6, 49% women, 92% STEMI, 0% fatal) were includedin our prospective study from 2010 to 2012. All the patients underwent successful primary percutaneous coronary intervention. MPV was determined on admission (MPV1) and in consecutive two days of hospitalization (MPV2, MPV3). The blood sample was analyzed immediately after collection in EDTA tubes using an automatic blood counter. Results: MPV1 was similar in both groups (8.91 ± 1.11 fl vs. 8.57 ± 0.74 fl, NS). Furthermore, there were no statistically significant differences in MPV value in fatal and nonfatal CS(8.90 ± 1.18 fl vs. 8.93 ± 1.05 fl, NS). Conclusions: The above results suggest that MPV cannot be considered a predictor of poorin-hospital outcome in patients with ACS complicated by cardiogenic shock.

Background: The study aimed to examine the relationship between vitamin D levels and the extent of coronary stenotic lesions in patients with ST-segment elevation myocardial infarction (STEMI). Experimental evidence points to the involvement of multiple factors in coronary plaque formation, including vitamin D. Little is known, however, about the association of vitamin D level with the intensity of atherosclerosis. Methods: Serum concentrations of 25-hydroxyvitamin D [25(OH)D] were measured in 130 consecutive patients with the first acute STEMI treated with primary percutaneous coronary intervention. STEMI was the first symptom of coronary artery disease (CAD). The study population was divided into patients with single and multi-vessel CAD. The angiographic severityof CAD was also determined based on the Gensini score. Results: The median 25(OH)D concentration was 11.12 ng/mL (25th and 75th percentile: 6.05; 17.12). Insufficient (20–30 ng/mL), deficient (10–20 ng/mL) and severely deficient(< 10 ng/mL) 25(OH)D levels were present in 18%, 35% and 45% of the individuals, respectively. Only 2 (2%) of patients had proper 25(OH)D levels (> 30 ng/mL). The 25(OH)D concentrations between patients with single- and multi-vessel CAD did not differ significantly (10.2 vs. 11.4 ng/mL, p = 0.62). There was no significant correlation between 25(OH)D levels and Gensini score (r = –0.0221, p = 0.81). Conclusions: The study demonstrated that vitamin D level is not associated with the severity of coronary lesions in patients with the first STEMI. A high prevalence of vitamin D deficiency in these patients was confirmed. Background: The study aimed to examine the relationship between vitamin D levels and the extent of coronary stenotic lesions in patients with ST-segment elevation myocardial infarction (STEMI). Experimental evidence points to the involvement of multiple factors in coronary plaque formation, including vitamin D. Little is known, however, about the association of vitamin D level with the intensity of atherosclerosis. Methods: Serum concentrations of 25-hydroxyvitamin D [25(OH)D] were measured in 130 consecutive patients with the first acute STEMI treated with primary percutaneous coronary intervention. STEMI was the first symptom of coronary artery disease (CAD). The study population was divided into patients with single and multi-vessel CAD. The angiographic severityof CAD was also determined based on the Gensini score. Results: The median 25(OH)D concentration was 11.12 ng/mL (25th and 75th percentile: 6.05; 17.12). Insufficient (20–30 ng/mL), deficient (10–20 ng/mL) and severely deficient(< 10 ng/mL) 25(OH)D levels were present in 18%, 35% and 45% of the individuals, respectively. Only 2 (2%) of patients had proper 25(OH)D levels (> 30 ng/mL). The 25(OH)D concentrations between patients with single- and multi-vessel CAD did not differ significantly (10.2 vs. 11.4 ng/mL, p = 0.62). There was no significant correlation between 25(OH)D levels and Gensini score (r = –0.0221, p = 0.81). Conclusions: The study demonstrated that vitamin D level is not associated with the severity of coronary lesions in patients with the first STEMI. A high prevalence of vitamin D deficiency in these patients was confirmed.