EGF receptor trafficking: consequences for signaling and cancer

The ligand-stimulated epidermal growth factor receptor (EGFR) has been extensively studied in the analysis of molecular mechanisms regulating endocytic traffic and the role of that traffic in signal transduction. Although such studies have largely focused on mitogenic signaling and dysregulated traffic in tumorigenesis, there is growing interest in the potential role of EGFR traffic in cell survival and the consequent response to cancer therapy. Here we review recent advances in our understanding of molecular mechanisms regulating ligand-stimulated EGFR activation, internalization, and post-endocytic sorting. The role of EGFR overexpression/mutation and new modulators of EGFR traffic in cancer and the response to cancer therapeutics are also discussed. Finally, we speculate on the relationship between EGFR traffic and cell survival

WASH and Tsg101/ALIX-dependent diversion of stress-internalized EGFR from the canonical endocytic pathway

Stress exposure triggers ligand-independent EGF receptor (EGFR) endocytosis, but its post-endocytic fate and role in regulating signalling are unclear. We show that the p38 MAP kinase-dependent, EGFR tyrosine kinase (TK)-independent EGFR internalization induced by ultraviolet light C (UVC) or the cancer therapeutic cisplatin, is followed by diversion from the canonical endocytic pathway. Instead of lysosomal degradation or plasma membrane recycling, EGFR accumulates in a subset of LBPA-rich perinuclear multivesicular bodies (MVBs) distinct from those carrying EGF-stimulated EGFR. Stress-internalized EGFR co-segregates with exogenously expressed pre-melanosomal markers OA1 and fibrillar PMEL, following early endosomal sorting by the actin polymerization-promoting WASH complex. Stress-internalized EGFR is retained intracellularly by continued p38 activity in a mechanism involving ubiquitin-independent, ESCRT/ALIX-dependent incorporation onto intraluminal vesicles (ILVs) of MVBs. In contrast to the internalization-independent EGF-stimulated activation, UVC/cisplatin-triggered EGFR activation depends on EGFR internalization and intracellular retention. EGFR signalling from this MVB subpopulation delays apoptosis and might contribute to chemoresistance

Paediatric nonbronchoscopic bronchoalveolar lavage: Overview and recommendations for clinical practice

NB-BAL is an effective procedure for the diagnosis of pulmonary disease processes in ventilated infants and children. This procedure is, however, not without risks to both patients and staff. Numerous complications of NB-BAL exist, with hypoxia being the most common. As a result, care should be taken in performing NB-BAL on haemodynamically unstable patients; patients with coagulation defects; and patients with cardiac or brain abnormalities. This paper presents an overview of paediatric nonbronchoscopic bronchoalveolar lavage (NB-BAL) including: the rationale for NB-BAL; the complications associated with the procedure; indications and contraindications. It also recommends an evidence-based clinical guideline for performing the procedure in the paediatric intensive care unit. By following the NB-BAL guidelines presented in this paper, one can ensure that an effective specimen is obtained from the lower respiratory tract, whilst minimising the risk to the patient

Annexin A1 Tethers Membrane Contact Sites that Mediate ER to Endosome Cholesterol Transport

Membrane contact sites between the ER and multivesicular endosomes/bodies (MVBs) play important roles in endosome positioning and fission and in neurite outgrowth. ER-MVB contacts additionally function in epidermal growth factor receptor (EGFR) tyrosine kinase downregulation by providing sites where the ER-localized phosphatase, PTP1B, interacts with endocytosed EGFR before the receptor is sorted onto intraluminal vesicles (ILVs). Here we show that these contacts are tethered by annexin A1 and its Ca2+-dependent ligand, S100A11, and form a subpopulation of differentially regulated contact sites between the ER and endocytic organelles. Annexin A1-regulated contacts function in the transfer of ER-derived cholesterol to the MVB when low-density lipoprotein-cholesterol in endosomes is low. This sterol traffic depends on interaction between ER-localized VAP and endosomal oxysterol-binding protein ORP1L, and is required for the formation of ILVs within the MVB and thus for the spatial regulation of EGFR signaling

Retinal pigment epithelial cells mitigate the effects of complement attack by endocytosis of C5b-9

Retinal pigment epithelial (RPE) cell death is a hallmark of age-related macular degeneration. The alternative pathway of complement activation is strongly implicated in RPE cell dysfunction and loss in age-related macular degeneration; therefore, it is critical that RPE cells use molecular strategies to mitigate the potentially harmful effects of complement attack. We show that the terminal complement complex C5b-9 assembles rapidly on the basal surface of cultured primary porcine RPE cells but disappears over 48 h without any discernable adverse effects on the cells. However, in the presence of the dynamin inhibitor dynasore, C5b-9 was almost completely retained at the cell surface, suggesting that, under normal circumstances, it is eliminated via the endocytic pathway. In support of this idea, we observed that C5b-9 colocalizes with the early endosome marker EEA1 and that, in the presence of protease inhibitors, it can be detected in lysosomes. Preventing the endocytosis of C5b-9 by RPE cells led to structural defects in mitochondrial morphology consistent with cell stress. We conclude that RPE cells use the endocytic pathway to prevent the accumulation of C5b-9 on the cell surface and that processing and destruction of C5b-9 by this route are essential for RPE cell survival

Influence motor aktivity on physical development of students

The character of organic carbon (OC) in lake waters is strongly dependent on the time water has spent in the landscape as well as in the lake itself due to continuous biogeochemical OC transformation processes. A common view is that upstream lakes might prolong the water retention in the landscape, resulting in an altered OC character downstream. We calculated the number of lakes upstream for 24,742 Swedish lakes in seven river basins spanning from 56º to 68º N. For each of these lakes, we used a lake volume to discharge comparison on a landscape scale to account for upstream water retention by lakes (Tn tot). We found a surprisingly weak relationship between the number of lakes upstream and Tn tot. Accordingly, we found that the coloured fraction of organic carbon was not related to lake landscape position but significantly related to Tn tot when we analysed lake water chemical data from 1,559 lakes in the studied river basins. Thus, we conclude that water renewal along the aquatic continuum by lateral water inputs offsets cumulative retention by lakes. Based on our findings, we suggest integrating Tn tot in studies that address lake landscape position in the boreal zone to better understand variations in the character of organic carbon across lake districts

PERSiST: a flexible rainfall-runoff modelling toolkit for use with the INCA family of models

Runoff generation processes and pathways vary widely between catchments. Credible simulations of solute and pollutant transport in surface waters are dependent on models which facilitate appropriate, catchment-specific representations of perceptual models of the runoff generation process. Here, we present a flexible, semi-distributed landscape-scale rainfall-runoff modelling toolkit suitable for simulating a broad range of user-specified perceptual models of runoff generation and stream flow occurring in different climatic regions and landscape types. PERSiST (the Precipitation, Evapotranspiration and Runoff Simulator for Solute Transport) is designed for simulating present-day hydrology; projecting possible future effects of climate or land use change on runoff and catchment water storage; and generating hydrologic inputs for the Integrated Catchments (INCA) family of models. PERSiST has limited data requirements and is calibrated using observed time series of precipitation, air temperature and runoff at one or more points in a river network. Here, we apply PERSiST to the river Thames in the UK and describe a Monte Carlo tool for model calibration, sensitivity and uncertainty analysi

The intercalated BSc (Med) Honours/MB ChB and integrated MB ChB/PhD tracks at the University of Cape Town: Models for a national medical student research training programme

The Faculty of Health Sciences at the University of Cape Town is addressing the shortage of clinician-scientists in South Africa by introducing two research training tracks in parallel with the professional MB ChB programme, namely the intercalated BSc (Med) Hons/MB ChB track and the integrated MB ChB/PhD track. The BSc (Med) Hons/MB ChB track is available to MB ChB students who have completed the first two years of study. The track comprises a course in Molecular Medicine given concurrently with the MB ChB third-year curriculum, followed by a BSc (Med) Hons as a ‘year out’ of MB ChB. Subsequently students may enrol into the integrated MB ChB/PhD track that enables them to undertake a PhD concurrently with MB ChB studies, which will be spread over additional years, or alternatively to undertake a PhD after completion of the MB ChB. These tracks, which were launched in 2011, represent an opportunity to train a new cadre of young African clinician-scientists at the undergraduate level.