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

Hydroxyapatite promotes superior adhesion and proliferation of telomerase transformed keratocytes in comparison with inert plastic skirt materials used in leading contemporary keratoprostheses

Aim: Published clinical series suggest the osteoodontokeratoprosthesis (OOKP) may have a lower extrusion rate than current synthetic keratoprostheses. The OOKP is anchored in the eye wall by autologous tooth. The authors’ aim was to compare adhesion, proliferation, and morphology for telomerase transformed keratocytes seeded on calcium hydroxyapatite (the principal mineral constituent of tooth) and materials used in the anchoring elements of commercially available synthetic keratoprostheses. Methods: Test materials were hydroxyapatite, polytetrafluoroethylene (PTFE), polyhydroxyethyl methacrylate (HEMA), and glass (control). Cell adhesion and viability were quantified at 4 hours, 24 hours, and 1 week using a calcein-AM/EthD-1 viability/cytotoxicity assay. Focal contact expression and cytoskeletal organisation were studied at 24 hours by confocal microscopy with immunoflourescent labelling. Further studies of cell morphology were performed using light and scanning electron microscopy. Results: Live cell counts were significantly greater on hydroxyapatite surfaces at each time point (p<0.04). Dead cell counts were significantly higher for PTFE at 7 days (p<0.002). Β1 integrin expression was highest on hydroxyapatite. Adhesion structures were well expressed in flat, spread out keratocytes on both HA and glass. Keratocytes tended to be thinner and spindle shaped on PTFE. The relatively few keratocytes visible on HEMA test surfaces were rounded and poorly adherent. Conclusions: Keratocyte adhesion, spreading, and viability on hydroxyapatite test surfaces is superior to that seen on PTFE and HEMA. Improving the initial cell adhesion environment in the skirt element of keratoprostheses may enhance tissue integration and reduce device failure rates

The impacts of future climate change and sulphur emission reductions on acidification recovery at Plastic Lake, Ontario

International audienceClimate-induced drought events have a significant influence on sulphate export from forested catchments in central Ontario, subsequently delaying the recovery of surface waters from acidification. In the current study, a model chain that employed a statistical downscaling model, a hydrological model and two hydrochemical models was used to forecast the chemical recovery of Plastic Lake sub-catchment 1 (PC1) from acidification under proposed deposition reductions and the A2 emission scenario of the Intergovernmental Panel on Climate Change. Any predicted recovery in stream acid neutralising capacity and pH owing to deposition reductions were clearly offset by large acid effluxes from climate-induced drought events. By 2100, ANC is predicted to show large variations ranging between 10 and ?30 ?molc L?1. Similarly, predicted pH in 2100 is lower (>0.05 of a pH unit) than the value simulated for 2000 (pH 4.35). Despite emission reductions, the future scenario paints a bleak picture of reacidification at PC1 to levels commensurate with those of the late 1970s. The principal process behind this reacidification is the oxidation of previously stored (reduced) sulphur compounds in wetlands during periods of low-flow (or drought), with subsequent efflux of sulphate upon re-wetting. Simulated catchment runoff under the A2 emissions scenario predictes increased intensity and frequency of low-flow events from approximately 2030 onwards. The Integrated Catchments model for Carbon indicated that stream DOC concentrations at PC1 will also increase under the future climate scenario, with temperature being the principal driver. Despite the predicted (significant) increase in DOC, pH is not predicted to further decline (beyond the climate-induced oxidation scenario), instead pH shows greater variability throughout the simulation. As echoed by many recent studies, hydrochemical models and model frameworks need to incorporate the drivers and mechanisms (at appropriate time-scales) that affect the key biogeochemical processes to reliably predict the impacts of climate change

Pulling the monstrosity of (hetro)normativity out of the closet: Teacher education as a problem and an answer

For the purposes of this chapter we suggest that (hetero )normativity¹ is a 'malaise' and a 'monstrous spectre, a menacing form of 'symbolic violence' in classrooms within universities, schools and early childhood centres. It is time to bring it out of the closet, not because it is hidden from view but because it is ubiquitous as a naturalised or taken-for-granted practice in the closets of our perceptions. As authors and activist teacher educators/academics/researchers working in a range of education settings, we are committed to changing the status quo by challenging this malaise and monstrosity. The question we face is: How might (hetero)normativity be exposed, challenged and addressed within initial teacher education (ITE) programmes? In this chapter we introduce and contextualise the issue of (hetero) normativity within our own ITE programmes, and identify some useful concepts. We provide a narrative that illustrates (hetero )normativity in education within Aotearoa/New Zealand. Writing in a manner so you, the reader, can 'perch on the periphery' and 'listen in' to the issues and dilemmas we are contending with in our work, we apply Bourdieu's theoretical framework to aid reflection. Our chapter concludes by offering some questions to consider how we, and you, might negotiate our situated practices to accost the spectre - that is, how we might expose and challenge heterosexuality and educate for positive change in relation to sexual diversity in schooling and teacher education

Modelling sediment supply and transport in the River Lugg: strategies for controlling sediment loads

The River Lugg has particular problems with high sediment loads that have resulted in detrimental impacts on ecology and fisheries. A new dynamic, process-based model of hydrology and sediments (INCA- SED) has been developed and applied to the River Lugg system using an extensive data set from 1995–2008. The model simulates sediment sources and sinks throughout the catchment and gives a good representation of the sediment response at 22 reaches along the River Lugg. A key question considered in using the model is the management of sediment sources so that concentrations and bed loads can be reduced in the river system. Altogether, five sediment management scenarios were selected for testing on the River Lugg, including land use change, contour tillage, hedging and buffer strips. Running the model with parameters altered to simulate these five scenarios produced some interesting results. All scenarios achieved some reduction in sediment levels, with the 40% land use change achieving the best result with a 19% reduction. The other scenarios also achieved significant reductions of between 7% and 9%. Buffer strips produce the best result at close to 9%. The results suggest that if hedge introduction, contour tillage and buffer strips were all applied, sediment reductions would total 24%, considerably improving the current sediment situation. We present a novel cost-effectiveness analysis of our results where we use percentage of land removed from production as our cost function. Given the minimal loss of land associated with contour tillage, hedges and buffer strips, we suggest that these management practices are the most cost-effective combination to reduce sediment loads

Hrs- and CD63-dependent competing mechanisms make different sized endosomal intraluminal vesicles

Multivesicular endosomes/bodies (MVBs) contain intraluminal vesicles (ILVs) that bud away from the cytoplasm. Multiple mechanisms of ILV formation have been identified, but the relationship between different populations of ILVs and MVBs remains unclear. Here we show in HeLa cells that different ILV subpopulations can be distinguished by size. EGF stimulation promotes the formation of large ESCRT-dependent ILVs, while depletion of the ESCRT-0 component, Hrs, promotes the formation of a uniformly sized population of small ILVs, the formation of which requires CD63. CD63 has previously been implicated in ESCRT-independent sorting of PMEL in MVBs and transfected PMEL is present on the small ILVs that form on Hrs depletion. Upregulation of CD63-dependent ILV formation by Hrs depletion indicates that Hrs and CD63 regulate competing machineries required for the generation of distinct ILV subpopulations. Taken together our results indicate that ILV size is influenced by their cargo and mechanism of formation and suggest a competitive relationship between ESCRT-dependent and -independent mechanisms of ILV formation within single MVBs