PhospholipaseA2: a key regulator of inflammatory signalling and a connector to fibrosis development in atherosclerosis

Atherosclerosis is a progressive inflammatory disease that takes place in the intima of the arterial wall. It is characterized by activation of endothelial cells, proliferation of smooth muscle cells and macrophages, accumulation of lipoproteins, deposition of extracellular matrix components and enhanced lipolytic enzyme activity. Phospholipase A(2) (PLA(2)) has been postulated to play an important role in the inflammatory process of atherosclerosis, but its molecular mechanism is uncertain. The secretory PLA(2) is expressed at increased levels in an atherosclerotic plaque and may hydrolyze low-density lipoproteins (LDL). This action promotes the production of pro-inflammatory lipids such as lysophospholipids, unsaturated fatty acids and eicosanoids. The current review highlights recent findings on how LDL-derived lipid mediators, generated by sPLA_2 modification of LDL, regulate pro-inflammatory activation and intracellular signaling in macrophages. Moreover, the review discusses how PLA_2 enzymes regulate signalling that promotes collagen accumulation and fibrotic plaque development. PLA_2 could therefore function as a connector between inflammation and fibrosis, the latter being an endpoint of chronic inflammation.     

EGFR and ADAMs cooperate to regulate shedding and endocytic trafficking of the desmosomal cadherin desmoglein 2

Regulation of classic cadherins plays a critical role in tissue remodeling during development and cancer; however, less attention has been paid to the importance of desmosomal cadherins. We previously showed that EGFR inhibition results in accumulation of the desmosomal cadherin, desmoglein 2 (Dsg2), at cell-cell interfaces accompanied by inhibition of matrix metalloprotease (MMP)-dependent shedding of the Dsg2 ectodomain and tyrosine phosphorylation of its cytoplasmic domain. Here, we show that EGFR inhibition stabilizes Dsg2 at intercellular junctions by interfering with its accumulation in an internalized cytoplasmic pool. Furthermore, MMP inhibition and ADAM17 RNAi, blocked shedding and depleted internalized Dsg2, but less so E-cadherin, in highly invasive SCC68 cells. ADAM9 and 15 silencing also impaired Dsg2 processing, supporting the idea that this desmosomal cadherin can be regulated by multiple ADAM family members. In contrast, ADAM10 siRNA enhanced accumulation of a 100-kDa Dsg2 cleavage product and internalized pool of Dsg2. Although both MMP and EGFR inhibition increased intercellular adhesive strength in control cells, the response to MMP-inhibition was Dsg2-dependent. These data support a role for endocytic trafficking in regulating desmosomal cadherin turnover and function and raise the possibility that internalization and regulation of desmosomal and classic cadherin function can be uncoupled mechanistically.     

Phosphoinositides: key players in cell signalling, in time and space

Over the last few years, many reports have extended our knowledge of the inositol lipid metabolism and brought out some exciting information about the location, the variety and the role of phosphoinositides (PIs). Besides the so-called "canonical PI pathway" leading to the production of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), the precursor of the intracellular second messengers inositol 1,4,5-trisphosphate and diacylglycerol (DAG), many other metabolic pathways have been identified to produce seven different polyphosphoinositides. Several of these quantitatively minor lipid molecules appear to be specifically involved in the control of cellular events, such as the spatial and temporal organisation of key signalling pathways, the rearrangement of the actin cytoskeleton or the intracellular vesicle trafficking. This is consistent with the fact that many of the enzymes, such as kinases and phosphatases, involved in the tight control of the intracellular level of polyphosphoinositides, are regulated and/or relocated through cell surface receptors for extracellular ligands. The remarkable feature of PIs, which can be rapidly synthesised and degraded in discrete membrane domains or even subnuclear structures, places them as ideal regulators and integrators of very dynamic mechanisms of cell regulation. In this review, we will summarise recent studies on the potential location, the metabolic pathways and the role of the different PIs. Some aspects of the temporal synthesis of D3 PIs will also be discussed.     

Endocytosis in signalling and development

After a long period of neglect, endocytosis in plants is finally coming of age. The constitutive recycling of plasma membrane proteins has been well established in the past few years, and recent studies report the ligand-induced endocytosis of receptors and other plasma membrane proteins. Signalling by ligand-bound receptors from endosomes has not, however, been demonstrated in plants. Although novel markers have been used to map endocytic pathways, the functional compartmentalisation of endosomes is still controversial. It is thus not clear where and how cargo proteins such as receptors are sorted towards either recycling to the plasma membrane or targeting to the vacuole for degradation.     

Regulation of EGFR signalling by palmitoylation and its role in tumorigenesis

The epidermal growth factor receptor (EGFR) is an essential driver of oncogenic signalling, and EGFR inhibitors are some of the earliest examples of successful targeted therapies in multiple types of cancer. The tractability of EGFR as a therapeutic target is overshadowed by the inevitable drug resistance that develops. Overcoming resistance mechanisms requires a deeper understanding of EGFR regulation in cancer cells. In this review, we discuss our recent discovery that the palmitoyltransferase DHHC20 palmitoylates EGFR on the C-terminal domain and plays a critical role in signal regulation during oncogenesis. Inhibiting DHHC20 expression or mutating the palmitoylation site on EGFR alters the EGF-induced signalling kinetics from a transient signal to a sustained signal. The change in signalling is accompanied by a decrease in cell proliferation in multiple human cancer cell lines. Our in vivo studies demonstrate that ablating the gene Zdhhc20 by CRISPR/Cas9-mediated inhibition in a mouse model of oncogenic Kras-driven lung adenocarcinoma potently inhibits tumorigenesis. The negative effect on tumorigenesis is mediated by EGFR since the expression of a palmitoylation-resistant mutant form of EGFR also inhibits Kras-driven lung adenocarcinoma. Finally, reducing EGFR palmitoylation increases the sensitivity of multiple cancer cell lines to existing inhibitors of EGFR and downstream signalling effector pathways. We will discuss the implications of these effects and strategies for targeting these new vulnerabilities.     

p130Cas: A key signalling node in health and disease

p130Cas/breast cancer anti-oestrogen resistance 1 (BCAR1) is a member of the Cas (Crk-associated substrate) family of adaptor proteins, which have emerged as key signalling nodes capable of interactions with multiple proteins, with important regulatory roles in normal and pathological cell function. The Cas family of proteins is characterised by the presence of multiple conserved motifs for protein–protein interactions, and by extensive tyrosine and serine phosphorylations. Recent studies show that p130Cas contributes to migration, cell cycle control and apoptosis. p130Cas is essential during early embryogenesis, with a critical role in cardiovascular development. Furthermore, p130Cas has been reported to be involved in the development and progression of several human cancers. p130Cas is able to perform roles in multiple processes due to its capacity to regulate a diverse array of signalling pathways, transducing signals from growth factor receptor tyrosine kinases, non-receptor tyrosine kinases, and integrins. In this review we summarise the current understanding of the structure, function, and regulation of p130Cas, and discuss the importance of p130Cas in both physiological and pathophysiological settings, with a focus on the cardiovascular system and cancer.     

The complexity of targeting EGFR signalling in cancer: from expression to turnover

The epidermal growth factor receptor (ErbB1 or EGFR) has been found to be altered in a variety of human cancers. A number of agents targeting these receptors, including specific antibodies directed against the ligand-binding domain of the receptor and small molecules that inhibit kinase activity are either in clinical trials or are already approved for clinical treatment. However, identifying patients that are likely to respond to such treatments has been challenging. As a consequence, it still remains important to identify additional alterations of the tumor cell that contribute to the response to EGFR-targeted agents. While EGFR-mediated signalling pathways have been well established, there is still a rather limited understanding of how intracellular protein-protein interactions, ubiquitination, endocytosis and subsequent degradation of EGFR contribute to the determination of sensitivity to EGFR targeting agents and are emerging areas of investigation. This review primarily focuses on the basic signal transduction pathways mediated through activated membrane bound and/or endosomal EGFR and emphasizes the need to co-target additional proteins that function either upstream or downstream of EGFR to improve cancer therapy.     

Feedback regulation of EGFR signalling: decision making by early and delayed loops

Human-made information relay systems invariably incorporate central regulatory components, which are mirrored in biological systems by dense feedback and feedforward loops. This type of system control is exemplified by positive and negative feedback loops (for example, receptor endocytosis and dephosphorylation) that enable growth factors and receptor Tyr kinases of the epidermal growth factor receptor (EGFR)/ERBB family to regulate cellular function. Recent studies show that the collection of feedback regulatory loops can perform computational tasks - such as decoding ligand specificity, transforming graded input signals into a digital output and regulating response kinetics. Aberrant signal processing and feedback regulation can lead to defects associated with pathologies such as cancer.     

Chewing the fat: beta-oxidation in signalling and development

Peroxisomal beta-oxidation is involved not only in fatty acid catabolism and lipid housekeeping but also in metabolism of hormones and amino acids in plants. Recent research in model species has led to new insights into the roles of this pathway in signalling and development, in particular regarding the involvement of beta-oxidation in jasmonic acid biosynthesis. Analysis of associated processes, such as the glyoxylate cycle and redox metabolism has also highlighted the importance of integration of beta-oxidation with cytosolic and mitochondrial metabolism. Mutations that disrupt beta-oxidation can have extremely pleiotropic effects, indicating important and varied roles for this pathway throughout the plant life cycle and making this an exciting topic for future research.     

Receptors and signalling components of plant hormones

Recent advances in understanding plant hormonal signalling has resulted in the identification of a variety of signalling components including receptor kinases with homology to the bacterial two component system as well as serine/threonine kinases and protein phosphatases. In addition, the existence of MAP kinase pathways in plants indicates a similar role of these signalling cascades in the relay of exogenous signals into the nucleus as has been disclosed in animal cells. The emerging signalling pathways of the plant hormone abscisic acid and ethylene are presented.     

Phosphatidyl-inositol 3-kinase: a key enzyme in diverse signalling processes

Phosphatidylinositol (PI) 3-kinase associates in signal-transducing complexes with activated growth factor receptors and other protein tyrosine kinases. The enzyme may also act downstream of receptors that are not tyrosine kinases in terminally differentiated cells. The recent cloning of the catalytic subunit of PI 3-kinase has revealed a structural similarity to a yeast protein important in vacuolar protein sorting. This finding provides some interesting clues to the function of PI3-kinase in diverse cellular responses.     

SNAREs--molecular governors in signalling and development

SNARE (soluble N-ethylmaleimide-sensitive factor protein attachment protein receptor) proteins drive membrane fusion and contribute to membrane and protein targeting and delivery in all eukaryotic cells. SNAREs are essential to the mechanics of cell growth and development, and they facilitate a number of homeostatic and evoked responses in plants, from hormone signalling to pathogen defence. Additionally, there is now unambiguous evidence that SNAREs play roles in anchoring other membrane proteins and in facilitating ion channel gating through direct, physical interaction with channel proteins. What is the physiological significance of these additional features of plant SNAREs? We explore possible interpretations and suggest functions as scaffolds for effective signal transmission between proteins and, by analogy with a mechanical device invented by James Watt, as molecular governors to coordinate solute transport with cell expansion and growth.