Georg Ende

Ephrin-A1/EphA4-mediated adhesion of monocytes to endothelial cells

The Eph receptors represent the largest family of receptor tyrosine kinases. Both Eph receptors and their ephrin ligands are cell-surface proteins, and they typically mediate cell-to-cell communication by interacting at sites of intercellular contact. The major aim of the present study was to investigate the involvement of EphA4-ephrin-A1 interaction in monocyte adhesion to endothelial cells, as this process is a crucial step during the initiation and progression of the atherosclerotic plaque. Immunohistochemical analysis of human atherosclerotic plaques revealed expression of EphA4 receptor and ephrin-A1 ligand in major cell types within the plaque. Short-time stimulation of endothelial cells with the soluble ligand ephrin-A1 leads to a fourfold increase in adhesion of human monocytes to endothelial cells. In addition, ephrin-A1 further increases monocyte adhesion to already inflamed endothelial cells. EphrinA1 mediates its effect on monocyte adhesion via the activated receptor EphA4. This ephrinA1/EphA4 induced process involves the activation of the Rho signaling pathway and does not require active transcription. Rho activation downstream of EphA4 leads to increased polymerization of actin filaments in endothelial cells. This process was shown to be crucial for the proadhesive effect of ephrin-A1. The results of the present study show that ephrin-A1-induced EphA4 forward signaling promotes monocyte adhesion to endothelial cells via activation of RhoA and subsequent stress-fiber formation by a non-transcriptional mechanism.

TNF-α-mediated adhesion of monocytes to endothelial cells—The role of ephrinA1

The ligand ephrin A1 is more often discussed to play a role in the development of the atherosclerotic plaque and in this context especially in the monocyte adhesion to endothelial cells. As tumor necrosis factor-α (TNF-α) is known to induce monocyte adhesion to endothelium and ephrin A1 expression, the present study focuses on the involvement of ephrin A1 in TNF-α-mediated monocyte adhesion. The analysis of different members of the Eph/ephrin system in TNF-α-treated human umbilical vein endothelial cells (HUVEC) revealed that especially ephrinA1 was found to be highly regulated by TNF-α compared to other members of the Eph family. This effect is also present in arterial endothelial cells from the umbilical artery and from the coronary artery. This regulation is dependent on NFκB-activation as shown by the expression of a constitutive-active IκB-mutant. By using siRNA-mediated silencing and adenoviral overexpression of ephrinA1 in HUVEC, the involvement of ephrinA1 in the TNF-α triggered monocyte adhesion to endothelial cells could be demonstrated. In addition, these results could be verified by quantitative adhesion measurement using atomic force microscopy-based single-cell force spectroscopy and under flow conditions. Furthermore, this effect is mediated via the EphA4 receptor. EphrinA1 does not influence the mRNA or protein expression of the adhesion receptors VCAM-1 and ICAM-1 in endothelial cells. However, the surface presentation of these adhesion receptors is modulated in an ephrinA1-dependent manner. In conclusion, these data demonstrate that ephrinA1 plays an important role in the TNF-α-mediated adhesion of monocytes to endothelial cells, which might be of great importance in the context of atherosclerosis.

EphrinB2/EphA4-mediated activation of endothelial cells increases monocyte adhesion

The membrane anchored ligand ephrinB2 belongs to the broad Eph/ephrin system and is able to activate different Eph receptors. The Eph receptors belong to the huge group of receptor-tyrosine kinases. Eph receptors as well as their corresponding ephrin ligands are cell-membrane attached proteins. Therefore, direct cell-cell contact is essentially for interaction. It is known that ephrinB2 plays a pivotal role in developmental and in tumour angiogenesis. Previous studies point to a crucial role of the EphA4-receptor in the process of monocyte adhesion. Since ephrinB2 is known as an interaction partner of EphA4, the aim of the present study was to investigate a possible interplay of EphA4-receptor with ephrinB2 during monocyte adhesion to the endothelium. As verified by bulk adhesion assays and atomic-force microscopy based single-cell force spectroscopy, temporary stimulation of endothelial cells from different sources with the soluble ligand ephrinB2 increased monocyte adhesion to endothelial cells. The proadhesive effect of ephrinB2 was independent of an active transcription, but is mediated via the Rho signaling pathway with subsequent modulation of the actin cytoskeleton. Furthermore, ephrinB2 mediated its impact on monocyte adhesion via the receptor EphA4 as shown by siRNA-mediated silencing. Interestingly, ephrinB2 was induced by TNF-α treatment. Silencing of ephrinB2 led to a lowering of the TNF-α mediated monocyte adhesion to endothelial cells. Furthermore, immunohistochemical staining of human atherosclerotic plaque revealed expression of ephrinB2 in macrophages. The results of the present study point to a crucial role of ephrinB2 induced EphA4 forward signaling in the context of monocyte adhesion to endothelial cells. This transcription-independent effect is mediated by Rho signaling induced actin-filament polymerization.

Regulation of endothelial migration and proliferation by ephrin-A1

Endothelial migration and proliferation are fundamental processes in angiogenesis and wound healing of injured or inflamed vessels. The present study aimed to investigate the regulation of the Eph/ephrin-system during endothelial proliferation and the impact of the ligand ephrin-A1 on proliferation and migration of human umbilical venous (HUVEC) and arterial endothelial cells (HUAEC). Endothelial cells that underwent contact inhibition showed a massive induction of ephrin-A1. In contrast, an injury to a confluent endothelial layer, associated with induction of migration and proliferation, showed reduced ephrin-A1 levels. In addition, reducing ephrin-A1 expression by siRNA led to increased proliferation, whereas the overexpression of ephrin-A1 led to decreased proliferative activity. Due to the fact that wound healing is a combination of proliferation and migration, migration was investigated in detail. First, classical wound-healing assays showed increased wound closure in both ephrin-A1 silenced and overexpressing cells. Live-cell imaging enlightened the underlying differences. Silencing of ephrin-A1 led to a faster but more disorientated migration. In contrast, ephrin-A1 overexpression did not influence velocity of the cells, but the migration was more directed in comparison to the controls. Additional analysis of EphA2-silenced cells showed similar results in terms of proliferation and migration compared to ephrin-A1 silenced cells. Detailed analysis of EphA2 phosphorylation on ligand-dependent phospho-site (Y588) and autonomous activation site (S897) revealed a distinct phosphorylation pattern. Furthermore, the endothelial cells ceased to migrate when they came in contact with an ephrin-A1 coated surface. Using a baculoviral-mediated expression system, ephrin-A1 silencing and overexpression was shown to modulate the formation of focal adhesions. This implicates that ephrin-A1 is involved in changes of the actin cytoskeleton which explains the alterations in migratory actions, at least in part. In conclusion, ephrin-A1 expression is regulated by cellular density and is itself a critical determinant of endothelial proliferation. According to current knowledge, ephrin-A1 seems to be remarkably involved in elementary processes of endothelial migration like cellular polarization, migratory direction and speed. These data support the notion that ephrin-A1 plays a pivotal role in basal mechanisms of re-endothelialization.

The successful use of the Impella RP after a long cardiopulmonary resuscitation and systemic thrombolytic therapy in a patient with a fulminant pulmonary embolism: the first case report

Abstract Acute massive pulmonary embolism (PE) can result in progressive cardiogenic shock, right heart failure, and respiratory failure requiring cardiopulmonary resuscitation (CPR). We report the case of a 56-year-old woman who required prolonged CPR secondary to a highly suspected massive PE and cardiogenic shock. After receiving preclinical thrombolytic therapy, the patient was transferred to the intensive care unit with ongoing CPR. Because of persistent haemodynamic instability and acute right ventricular failure, an Impella RP was successfully implanted and immediate haemodynamic improvement was observed. Absent any contraindications, the Impella RP should be considered a feasible alternative in patients with acute right ventricular failure due to pulmonary embolism.

Elevated cardiac troponin T indicating myocardial infarction or myocarditis-urgent cardiac catheter examination-or do we have to consider other reasons?

Alveolar rhabdomyosarcoma is an aggressive tumour in adulthood, in which cardiac troponin T seems to be a tumour marker and course parameter. We present the clinical course of a young man suffering from this rare disease and the development of troponin T during therapy. Noninvasive cardiac imaging was used to exclude cardiac involvement, myocardial infarction or inflammation processes.

Acute coronary syndrome and single coronary artery ostium

We report the case of a 44-year-old man with a medical history of arterial hypertension and nicotine abuse (10 pack years) presenting with persistent chest pain to our emergency department. Cardiac markers were elevated (hsTrop T 112 ng/L; CK 4.33 μkat/L; CK-MB 0.67 μkat/L) and eletrocardiogram impressed with biphasic T waves in lead V1 to V4 (Fig. 1). Two-dimensional Doppler echocardiography showed normal biventricular function without regional wall motion disturbances. Left heart catheterisation revealed a single coronary artery from the left sinus valsalva with a right coronary artery (RCA) arising from the left main stem (LMS). Furthermore, a high grade stenosis of proximal left anterior descending (LAD) was displayed and percutaneous coronary intervention with implantation of 2 drug eluting stents was successfully performed (Figs. 2, 3). To document the extent of myocardial infarction, and to exclude a malignant course of RCA between aorta and pulmonary artery, the patient was referred to 3T cardiac magnetic resonance (MR) with an 8-channel cardiac coil. Late gadolinium enhancement (LGE) sequences presented small septal subendocardial contrast enhancement indicating myocardial fibrosis after myocardial infarction (Fig. 4). Using a 3D whole heart Fat Sat FIESTA technique sequence, RCA arising from the LMS was confirmed. But because of the 3T MR-specific low spatial resolution, RCA course could not be traced accurately (Fig. 5). Therefore, a coronary 128-slice dual-source computed tomography (CT) was carried out showing RCA running between aorta, left and right atrium and not interarterially (Figs. 6–8). Further conservative course of the patient proceeded without complications. This case demonstrates an isolated single coronary artery as a rare congenital anomaly occurring with an incidence of 0.02%. Although often being asymptomatic, this anomaly can also appear with a sudden cardiac event, as in this patient, or with a malignant course. Therefore, the necessity for multimodal imaging to illustrate the true anatomic conditions should be underlined. Additionally, the still existing difficulties of 3T MR coronary angiography in daily practice and the resolution advantages of CT in imaging coronary arteries were highlighted.