Junctional Adhesion Molecules (JAMs) - New Players in Breast Cancer?

Worldwide, breast cancer remains a leading cause of death amongst women. Annually, it i sestimated that breast cancer is diagnosed in over a million women (Kasler et al., 2009) with over 450,000 deaths worldwide (Tirona et al., 2010). The incidence of the disease is highest in economically-developed countries, with lower rates in developing countries. Despite continual advances in breast cancer care which have led to reduced mortality, however, the incidence of the disease is still rising. The decrease in breast cancer-specific mortality has been attributed to improvements in screening techniques which permit earlier detection, surgical and radiotherapy interventions, better understanding of disease pathogenesis and utilization of traditional chemotherapies in a more efficacious manner. Consequently, early stage breast cancer is now a curable disease while advanced breast cancer remains asignificant clinical problem. Breast cancer is a heterogeneous disease encompassing many subtypes, which differ both in terms of their molecular backgrounds and clinical prognosis. These breast cancer subtypes range from pre-invasive early stage disease to advanced invasive disease. The simplest classifications of disease subdivide breast cancer into pre-invasive and invasive forms; with the pre-invasive forms being ductal carcinoma in situ (DCIS) and lobular carcinoma in situ (LCIS). Carcinoma in situ is proliferation of cancer cells within the epithelial tissue without invasion of the surrounding stromal tissue (Bland & Copeland, 1998). DCIS arises in the terminal ductal lobular units (TDLU) and in extra-lobular ducts while LCIS occurs in the breast lobules, and is recognisable histopathologically by the presence of populations of aberrant cells with small nuclei (Hanby & Hughes, 2008). Invasive breast cancers are subclassified into invasive ductal breast cancer, invasive lobular breast cancer, inflammatory breast cancer and Paget's disease. Invasive ductal carcinoma (IDC) is the most common formof invasive breast cancer, accounting for around 85% of all cases. DCIS is frequently considered as an obligate precursor to IDC, progressing from lower to higher grades and then onto invasive cancer with progressive accumulation of genomic changes (Farabegoli et al ., 2002). However it has alternately been suggested that there exist genetically-distinct subgroups of DCIS, only some of which have the potential to progress to invasion (Shackney & Silverman, 2003). Long-term natural history studies of DCIS have provided supportive evidence for both possibilities (Page et al., 1995; Collins et al ., 2005; Sanders et al ., 2005). Despite such controversies, the large extent to which the genome is altered in DCIS strongly suggests that genomic instability precedes phenotypic evidence of invasion (Hwang et al ., 2004). This serves to underline the fact that malignant transformation in a heterogeneous disease like breast cancer is a dynamic process evolving through multiple multi-step pathway models. Many factors are thought to be responsible for the development of breast cancer. Genetic factors play a vital role in the predisposition to breast cancer, with mutations of BRCA1 and BRCA2 genes accounting for 5–10% of breast cancer cases and being responsible for 80% of inherited breast cancers (Nathanson et al., 2001). On a more complex level, much insight has been gained from the genetic profiling of thousands of tumours to generate gene signatures of prognostic value (Sorlie et al., 2001; van 't Veer et al., 2002; van de Vijver et al ., 2002), which have spurred the development of commercially-available diagnostic tests. The importance of reproductive factors in the aetiology of breast cancer is also well recognised with early onset of menarche, nulliparity, late menopause, endogenous and exogenous hormones representing the main risk factors (Reeves et al ., 2000; Key et al., 2001; Howell & Evans, 2011). Several other studies have reported anincreased risk of breast cancer with lack of physical activity (especially in pre menopausal women), as well as increasing age and obesity (Clarke et al ., 2006; Walker & Martin, 2007; Harrison et al., 2009; Rod et al., 2009; Awatef et al ., 2011). These risk factors accentuate the abnormal growth control of cells by increasing the circulating levels of oestrogen thereby promoting tumourigenesis within the breast microenvironment. A proper understandingof the breast cancer microenvironment is essential for understanding breast cancer, and will be explored in detail in the next sections. </p

Tight junctions: a barrier to the initiation and progression of breast cancer?

Breast cancer is a complex and heterogeneous disease that arises from epithelial cells lining the breast ducts and lobules. Correct adhesion between adjacent epithelial cells is important in determining the normal structure and function of epithelial tissues, and there is accumulating evidence that dysregulated cell-cell adhesion is associated with many cancers. This review will focus on one cell-cell adhesion complex, the tight junction (TJ), and summarize recent evidence that TJs may participate in breast cancer development or progression. We will first outline the protein composition of TJs and discuss the functions of the TJ complex. Secondly we will examine how alterations in these functions might facilitate breast cancer initiation or progression; by focussing on the regulatory influence of TJs on cell polarity, cell fate and cell migration. Finally we will outline how pharmacological targeting of TJ proteins may be useful in limiting breast cancer progression. Overall we hope to illustrate that the relationship between TJ alterations and breast cancer is a complex one; but that this area offers promise in uncovering fundamental mechanisms linked to breast cancer progression

Breast cancer cell migration is regulated through junctional adhesion molecule-A-mediated activation of Rap1 GTPase

ABSTRACT: INTRODUCTION: The adhesion protein junctional adhesion molecule-A (JAM-A) regulates epithelial cell morphology and migration, and its over-expression has recently been linked with increased risk of metastasis in breast cancer patients. As cell migration is an early requirement for tumor metastasis, we sought to identify the JAM-A signalling events regulating migration in breast cancer cells. METHODS: MCF7 breast cancer cells (which express high endogenous levels of JAM-A) and primary cultures from breast cancer patients were used for this study. JAM-A was knocked down in MCF7 cells using siRNA to determine the consequences for cell adhesion, cell migration and the protein expression of various integrin subunits. As we had previously demonstrated a link between the expression of JAM-A and β1-integrin, we examined activation of the β1-integrin regulator Rap1 GTPase in response to JAM-A knockdown or functional antagonism. To test whether JAM-A, Rap1 and β1-integrin lie in a linear pathway, we tested functional inhibitors of all three proteins separately or together in migration assays. Finally we performed immunoprecipitations in MCF7 cells and primary breast cells to determine the binding partners connecting JAM-A to Rap1 activation. RESULTS: JAM-A knockdown in MCF7 breast cancer cells reduced adhesion to, and migration through, the β1-integrin substrate fibronectin. This was accompanied by reduced protein expression of β1-integrin and its binding partners αV- and α5-integrin. Rap1 activity was reduced in response to JAM-A knockdown or inhibition, and pharmacological inhibition of Rap1 reduced MCF7 cell migration. No additive anti-migratory effect was observed in response to simultaneous inhibition of JAM-A, Rap1 and β1-integrin, suggesting that they lie in a linear migratory pathway. Finally, in an attempt to elucidate the binding partners putatively linking JAM-A to Rap1 activation, we have demonstrated the formation of a complex between JAM-A, AF-6 and the Rap1 activator PDZ-GEF2 in MCF7 cells and in primary cultures from breast cancer patients. CONCLUSIONS: Our findings provide compelling evidence of a novel role for JAM-A in driving breast cancer cell migration via activation of Rap1 GTPase and β1-integrin. We speculate that JAM-A over-expression in some breast cancer patients may represent a novel therapeutic target to reduce the likelihood of metastasis

Natural compound Tetrocarcin-A downregulates Junctional Adhesion Molecule-A in conjunction with HER2 and inhibitor of apoptosis proteins and inhibits tumor cell growth.

Overexpression of the tight junction protein Junctional Adhesion Molecule-A (JAM-A) has been linked to aggressive disease in breast and other cancers, but JAM-targeting drugs remain elusive. Screening of a natural compound library identified the antibiotic Tetrocarcin-A as a novel downregulator of JAM-A and human epidermal growth factor receptor-2 (HER2) protein expression in breast cancer cells. Lysosomal inhibition partially rescued the downregulation of JAM-A and HER2 caused by Tetrocarcin-A, and attenuated its cytotoxic activity. Tetrocarcin-A treatment or JAM-A silencing reduced AKT and ERK phosphorylation, inhibited c-FOS phosphorylation at Threonine-232 (its transcriptional regulation site), inhibited nuclear localization of c-FOS, and downregulated expression of the inhibitor of apoptosis proteins (IAP). This was accompanied by Tetrocarcin-A-induced caspase-dependent apoptosis. To begin evaluating the potential clinical relevance of our findings, we extended our studies to other models. Encouragingly, Tetrocarcin-A downregulated JAM-A expression and caused cytotoxicity in primary breast cells and lung cancer stem cells, and inhibited the growth of xenografts in a semi-in vivo model involving invasion across the chicken egg chorioallantoic membrane. Taken together, our data suggest that Tetrocarcin-A warrants future evaluation as a novel cancer therapeutic by virtue of its ability to downregulate JAM-A expression, reduce tumorigenic signaling and induce apoptosis

Duration of antibiotic treatment for common infections in English primary care: cross sectional analysis and comparison with guidelines

Objectives: To evaluate antibiotic therapy durations for common infections in English primary care and to compare this with guidelines. Design: Cross-sectional study. Setting: General practices contributing to The Health Improvement Network database, 2013-2015. Participants: 931,015 consultations that resulted in an antibiotic prescription for one of the following indications: acute sinusitis, acute sore throat, acute cough and bronchitis, pneumonia, acute exacerbation of chronic obstructive pulmonary disease (COPD), acute otitis media, acute cystitis, prostatitis, pyelonephritis, cellulitis, impetigo, scarlet fever and gastroenteritis. Main outcome measures: The main outcomes were the percentage of antibiotic prescriptions with a duration exceeding the guideline recommendation and the total number of days beyond the recommended duration for each indication. Results: The most common reasons for the prescriptions were patients consulting with acute bronchitis and cough (386,972), acute sore throat (239,231), acute otitis media (83,054), and acute sinusitis (76,683). Antibiotic treatments for upper respiratory indications and acute bronchitis accounted for more than two thirds of the total prescriptions considered, and ≥80% of these treatment courses exceeded guideline recommendations. Notable exceptions were acute sinusitis, where only 9.6% (95% CI 9.4 to 9.9%) of prescriptions exceeded 7 days and acute sore throat where only 2.1% (95% CI 2.0 to 2.1) exceed 10 days (recent guidance recommends 5 days). More than half of antibiotic prescriptions were longer than guidelines recommend for acute cystitis among females (54.6%, 95% CI 54.1 to 55.0%). The percentage of antibiotic prescriptions exceeding the recommended duration was lower for most non-respiratory infections. For the 931,015 included consultations resulting in antibiotic prescriptions, approximately 1.3 million days were beyond the durations recommended by the guidelines. Conclusion: For most common infections treated in primary care, a substantial proportion of antibiotic prescriptions have durations exceeding those recommended in guidelines. Substantial reductions in antibiotic exposure can be accomplished by aligning antibiotic prescription durations with guidelines

Myosin II regulates the shape of three-dimensional intestinal epithelial cysts.

The development of luminal organs begins with the formation of spherical cysts composed of a single layer of epithelial cells. Using a model three-dimensional cell culture, this study examines the role of a cytoskeletal motor, myosin II, in cyst formation. Caco-2 and SK-CO15 intestinal epithelial cells were embedded into Matrigel, and myosin II was inhibited by blebbistatin or siRNA-mediated knockdown. Whereas control cells formed spherical cysts with a smooth surface, inhibition of myosin II induced the outgrowth of F-actin-rich surface protrusions. The development of these protrusions was abrogated after inhibition of F-actin polymerization or of phospholipase C (PLC) activity, as well as after overexpression of a dominant-negative ADF/cofilin. Surface protrusions were enriched in microtubules and their formation was prevented by microtubule depolymerization. Myosin II inhibition caused a loss of peripheral F-actin bundles and a submembranous extension of cortical microtubules. Our findings suggest that inhibition of myosin II eliminates the cortical F-actin barrier, allowing microtubules to reach and activate PLC at the plasma membrane. PLC-dependent stimulation of ADF/cofilin creates actin-filament barbed ends and promotes the outgrowth of F-actin-rich protrusions. We conclude that myosin II regulates the spherical shape of epithelial cysts by controlling actin polymerization at the cyst surface

In Vivo Flow Dynamics of the Total Cavopulmonary Connection from Three-dimensional Multislice Magnetic Resonance Imaging

Background. The total cavopulmonary connection (TCPC) design continues to be refined on the basis of flow analysis at the connection site. These refinements are of importance for myocardial energy conservation in the univentricular supported circulation. In vivo magnetic resonance phase contrast imaging provides semiquantitative flow visualization information. The purpose of this study was to understand the in vivo TCPC flow characteristics obtained by magnetic resonance phase contrast imaging and compare the results with our previous in vitro TCPC flow experiments in an effort to further refine TCPC surgical design. Methods. Twelve patients with TCPC underwent sedated three-dimensional, multislice magnetic resonance phase contrast imaging. Seven patients had intraatrial lateral tunnel TCPC and 5 had extracardiac TCPC. Results. In all patients in both groups a disordered flow pattern was observed in the inferior caval portion of the TCPC. Flow at the TCPC site appeared to be determined by connection geometry, being streamlined at the superior vena cava–pulmonary junction when the superior vena cava was offset and flared toward the left pulmonary artery. Without caval offset, intense swirling and dominance of superior vena caval flow was observed. In TCPC with bilateral superior vena cavae, the flow patterns observed included secondary vortices, a central stagnation point, and influx of the superior vena cava flow into the inferior caval conduit. A comparative analysis of in vivo flow and our previous in vitro flow data from glass model prototypes of TCPC demonstrated significant similarities in flow disturbances. Three-dimensional magnetic resonance phase contrast imaging in multiple coronal planes enabled a comprehensive semiquantitative flow analysis. The data are presented in traditional instantaneous images and in animated format for interactive display of the flow dynamics. Conclusions. Flow in the inferior caval portion of the TCPC is disordered, and the TCPC geometry determines flow characteristics

In Vivo Flow Dynamics of the Total Cavopulmonary Connection from Three-dimensional Multislice Magnetic Resonance Imaging

Background. The total cavopulmonary connection (TCPC) design continues to be refined on the basis of flow analysis at the connection site. These refinements are of importance for myocardial energy conservation in the univentricular supported circulation. In vivo magnetic resonance phase contrast imaging provides semiquantitative flow visualization information. The purpose of this study was to understand the in vivo TCPC flow characteristics obtained by magnetic resonance phase contrast imaging and compare the results with our previous in vitro TCPC flow experiments in an effort to further refine TCPC surgical design. Methods. Twelve patients with TCPC underwent sedated three-dimensional, multislice magnetic resonance phase contrast imaging. Seven patients had intraatrial lateral tunnel TCPC and 5 had extracardiac TCPC. Results. In all patients in both groups a disordered flow pattern was observed in the inferior caval portion of the TCPC. Flow at the TCPC site appeared to be determined by connection geometry, being streamlined at the superior vena cava–pulmonary junction when the superior vena cava was offset and flared toward the left pulmonary artery. Without caval offset, intense swirling and dominance of superior vena caval flow was observed. In TCPC with bilateral superior vena cavae, the flow patterns observed included secondary vortices, a central stagnation point, and influx of the superior vena cava flow into the inferior caval conduit. A comparative analysis of in vivo flow and our previous in vitro flow data from glass model prototypes of TCPC demonstrated significant similarities in flow disturbances. Three-dimensional magnetic resonance phase contrast imaging in multiple coronal planes enabled a comprehensive semiquantitative flow analysis. The data are presented in traditional instantaneous images and in animated format for interactive display of the flow dynamics. Conclusions. Flow in the inferior caval portion of the TCPC is disordered, and the TCPC geometry determines flow characteristics

Galaxy Star Formation as a Function of Environment in the Early Data Release of the Sloan Digital Sky Survey

We present in this paper a detailed analysis of the effect of environment on the star formation activity of galaxies within the Early Data Release (EDR) of the Sloan Digital Sky Survey (SDSS). We have used the Halpha emission line to derive the star formation rate (SFR) for each galaxy within a volume-limited sample of 8598 galaxies with 0.05 less than or equal to z less than or equal to 0.095 and M (r*) less than or equal to 20.45. We find that the SFR of galaxies is strongly correlated with the local ( projected) galaxy density, and thus we present here a density-SFR relation that is analogous to the density-morphology relation. The effect of density on the SFR of galaxies is seen in three ways. First, the overall distribution of SFRs is shifted to lower values in dense environments compared with the field population. Second, the effect is most noticeable for the strongly star-forming galaxies (Halpha EW &gt; 5 Angstrom) in the 75th percentile of the SFR distribution. Third, there is a break ( or characteristic density) in the density-SFR relation at a local galaxy density of similar to1 h(75)(-2) Mpc(-2). To understand this break further, we have studied the SFR of galaxies as a function of clustercentric radius from 17 clusters and groups objectively selected from the SDSS EDR data. The distribution of SFRs of cluster galaxies begins to change, compared with the field population, at a clustercentric radius of 3-4 virial radii (at the &gt;1sigma statistical significance), which is consistent with the characteristic break in density that we observe in the density-SFR relation. This effect with clustercentric radius is again most noticeable for the most strongly star-forming galaxies. Our tests suggest that the density-morphology relation alone is unlikely to explain the density-SFR relation we observe. For example, we have used the ( inverse) concentration index of SDSS galaxies to classify late-type galaxies and show that the distribution of the star-forming (EW Halpha &gt; 5Angstrom) late-type galaxies is different in dense regions ( within 2 virial radii) compared with similar galaxies in the field. However, at present, we are unable to make definitive statements about the independence of the density-morphology and density-SFR relation. We have tested our work against potential systematic uncertainties including stellar absorption, reddening, SDSS survey strategy, SDSS analysis pipelines, and aperture bias. Our observations are in qualitative agreement with recent simulations of hierarchical galaxy formation that predict a decrease in the SFR of galaxies within the virial radius. Our results are in agreement with recent 2dF Galaxy Redshift Survey results as well as consistent with previous observations of a decrease in the SFR of galaxies in the cores of distant clusters. Taken together, these works demonstrate that the decrease in SFR of galaxies in dense environments is a universal phenomenon over a wide range in density (from 0.08 to 10 h(75)(-2) Mpc(-2)) and redshift (out to z similar or equal to 0.5)

The Bathypelagic Biome of the Atlantic Ocean: Character and Ecological Discreteness of the Fish Fauna

Recent global synthetic analyses have revealed that marine taxonomic inventories are far from complete, nowhere more so than in the deep-pelagic ocean. At over a billion km3, it is the largest biome on Earth, yet only a tiny fraction of the biogeographic records include the bathypelagic fauna. This data gap served as the impetus for recent deepwater surveys, many of which have altered our perceptions of pelagic ecosystems. Here we examine data from four deep-pelagic (0-5000+ m) sampling programs in the Atlantic (60°N-25°S) in order to assess the character of bathypelagic fish communities with respect to faunal distinctiveness and ecological connectivity. Regions studied include the Gulf of Mexico, Sargasso 702 Sea, eastern North/South Atlantic, and mid-North Atlantic. Quantitative analyses give contrasting pictures with respect to faunal composition and ecosystem operation. The discreteness of the bathypelagic zone is exhibited faunistically by the suite of ―holobathypelagic‖ species found only below 1000 m, most of which are highly modified morphologically. Geometric abundance class analyses reveal that the character of relative species abundance distributions between the meso- and bathypelagic zones is fundamentally dissimilar; the former exhibit a much higher proportion of common species, while the latter exhibit a much higher percentage of rarer species. From a community energetics perspective, however, the bathy- and mesopelagic zones are highly interconnected. Approximately 70% of fish species collected below 1000 m are also found in the mesopelagic zone, and in the far North Atlantic, are also found in the epipelagial. These species comprised 66 to \u3e90% of individuals collected below 1000 m in the regions sampled. In the mid-North Atlantic, these species contribute to the unexpected water-column biomass maximum observed between 1500-2300 m. Thus, the ―transient‖ taxa (primarily mesopelagic migrators and spanner taxa) add considerably to the ichthyofaunal diversity of the world ocean below 1000 m, and appear to be the vectors that support the diverse array of holobathypelagic fishes whose taxonomic composition is dominated by piscivores. Data from the four regions studied suggests that classic pelagic biogeographic boundaries do not apply to bathypelagic realm, as shared species are the rule rather than the exception. Last, cumulative species curves suggest we are far from understanding the true complexity of the bathypelagic zone