Evaluation of the current knowledge limitations in breast cancer research: a gap analysis

BACKGROUND A gap analysis was conducted to determine which areas of breast cancer research, if targeted by researchers and funding bodies, could produce the greatest impact on patients. METHODS Fifty-six Breast Cancer Campaign grant holders and prominent UK breast cancer researchers participated in a gap analysis of current breast cancer research. Before, during and following the meeting, groups in seven key research areas participated in cycles of presentation, literature review and discussion. Summary papers were prepared by each group and collated into this position paper highlighting the research gaps, with recommendations for action. RESULTS Gaps were identified in all seven themes. General barriers to progress were lack of financial and practical resources, and poor collaboration between disciplines. Critical gaps in each theme included: (1) genetics (knowledge of genetic changes, their effects and interactions); (2) initiation of breast cancer (how developmental signalling pathways cause ductal elongation and branching at the cellular level and influence stem cell dynamics, and how their disruption initiates tumour formation); (3) progression of breast cancer (deciphering the intracellular and extracellular regulators of early progression, tumour growth, angiogenesis and metastasis); (4) therapies and targets (understanding who develops advanced disease); (5) disease markers (incorporating intelligent trial design into all studies to ensure new treatments are tested in patient groups stratified using biomarkers); (6) prevention (strategies to prevent oestrogen-receptor negative tumours and the long-term effects of chemoprevention for oestrogen-receptor positive tumours); (7) psychosocial aspects of cancer (the use of appropriate psychosocial interventions, and the personal impact of all stages of the disease among patients from a range of ethnic and demographic backgrounds). CONCLUSION Through recommendations to address these gaps with future research, the long-term benefits to patients will include: better estimation of risk in families with breast cancer and strategies to reduce risk; better prediction of drug response and patient prognosis; improved tailoring of treatments to patient subgroups and development of new therapeutic approaches; earlier initiation of treatment; more effective use of resources for screening populations; and an enhanced experience for people with or at risk of breast cancer and their families. The challenge to funding bodies and researchers in all disciplines is to focus on these gaps and to drive advances in knowledge into improvements in patient care

Overexpression of the oncostatin-M receptor in cervical squamous cell carcinoma is associated with epithelial-mesenchymal transition and poor overall survival.

BACKGROUND: Copy-number gain of the oncostatin-M receptor (OSMR) occurs frequently in cervical squamous cell carcinoma (SCC) and is associated with adverse clinical outcome. We previously showed that OSMR overexpression renders cervical SCC cells more sensitive to the major ligand oncostatin-M (OSM), which increases migration and invasion in vitro. We hypothesised that a major contribution to this phenotype would come from epithelial-mesenchymal transition (EMT). METHODS: We performed a comprehensive integrated study, involving in vitro cell line studies, in vivo animal models and numerous clinical samples from a variety of anatomical sites. RESULTS: In independent sets of cervical, head/neck and lung SCC tissues, OSMR expression levels correlated with multiple EMT-associated phenotypic markers and transcription factors. OSM treatment of OSMR overexpressing cervical SCC cells produced consistent EMT changes and increased tumour sphere formation in suspension culture. In a mouse model, OSMR overexpressing SCC cells treated with OSM showed significant increases in lung colonisation. The biological effects of exogenous OSM were mirrored by highly significant adverse overall survival in cervical SCCs with OSMR overexpression (N=251). CONCLUSIONS: OSM:OSMR interactions are able to induce EMT, increased cancer stem cell-like properties and enhanced lung colonisation in SCC cells. These changes are likely to contribute to the highly significant adverse outcome associated with OSMR overexpression in cervical SCCs.This work was supported by Cancer Research UK (Programme Grant A13080).This is the author accepted manuscript. It is currently under an indefinite embargo pending publication by Nature Publishing Group

The genetic diversity, phylogeography and morphology of Elphidiidae (Foraminifera) in the Northeast Atlantic

Genetic characterisation (SSU rRNA genotyping) and Scanning ElectronMicroscope (SEM) imaging of individualtests were used in tandem to determine the modern species richness of the foraminiferal family Elphidiidae(Elphidium, Haynesina and related genera) across the Northeast Atlantic shelf biomes. Specimens were collectedat 25 locations fromthe High Arctic to Iberia, and a total of 1013 individual specimenswere successfully SEMimagedand genotyped. Phylogenetic analyses were carried out in combination with 28 other elphidiid sequencesfrom GenBank and seventeen distinct elphidiid genetic types were identified within the sample set, sevenbeing sequenced for the first time. Genetic types cluster into sevenmain cladeswhich largely represent their generalmorphologicalcharacter. Differences between genetic types at the genetic, morphological and biogeographiclevels are indicative of species level distinction. Their biogeographic distributions, in combination with elphidiidSSU sequences from GenBank and high resolution images from the literature show that each of them exhibitsspecies-specific rather than clade-specific biogeographies. Due to taxonomic uncertainty and divergent taxonomicconcepts between schools, we believe that morphospecies names should not be placed onto molecularphylogenies unless both the morphology and genetic type have been linked to the formally named holotype,or equivalent. Based on strictmorphological criteria,we advocate using only a three-stage approach to taxonomyfor practical application in micropalaeontological studies. It comprises genotyping, the production of a formalmorphological description of the SEM images associated with the genetic type and then the allocation of themost appropriate taxonomic name by comparison with the formal type description. Using this approach, wewere able to apply taxonomic names to fifteen genetic types. One of the remaining two may be potentially cryptic,and one is undescribed in the literature. In general, the phylogeographic distribution is in agreement with ourknowledge of the ecology and biogeographical distribution of the corresponding morphospecies, highlighting thegenerally robust taxonomic framework of the Elphidiidae in time and space

Epithelial plasticity, stemness and pluripotency

3 páginas, 1 figura.Work in the lab is supported by grants BFU2008-01042, CONSOLIDER- INGENIO 2010 CSD2007-00017, CSD2007-00023 and Prometeo 2008/049 to MAN.Peer reviewe

Hepatitis C Virus Core Protein Inhibits Interferon Production by a Human Plasmacytoid Dendritic Cell Line and Dysregulates Interferon Regulatory Factor-7 and Signal Transducer and Activator of Transcription (STAT) 1 Protein Expression

<div><p>Plasmacytoid Dendritic Cells (pDCs) represent a key immune cell population in the defense against viruses. pDCs detect viral pathogen associated molecular patterns (PAMPs) through pattern recognition receptors (PRR). PRR/PAMP interactions trigger signaling events that induce interferon (IFN) production to initiate local and systemic responses. pDCs produce Type I and Type III (IFNL) IFNs in response to HCV RNA. Extracellular HCV core protein (Core) is found in the circulation in chronic infection. This study defined how Core modulates PRR signaling in pDCs. Type I and III IFN expression and production following exposure to recombinant Core or β-galactosiade was assessed in human GEN2.2 cells, a pDC cell line. Core suppressed type I and III IFN production in response to TLR agonists and the HCV PAMP agonist of RIG-I. Core suppression of IFN induction was linked with decreased IRF-7 protein levels and increased non-phosphorylated STAT1 protein. Circulating Core protein interferes with PRR signaling by pDCs to suppress IFN production. Strategies to define and target Core effects on pDCs may serve to enhance IFN production and antiviral actions against HCV.</p></div

rCore inhibits TLR stimulation.

<p>Gene fold increases in GEN2.2-pDCs following rCore pretreatment and TLR stimulation with Loxoribine (A) or CpGA (B). C) Kinetics of IFN mRNA after pretreatment with rCore or β-galactosidase (β-gal) and HCV pU/UC RNA stimulation. D) Levels of protein by ELISA when cells were treated with rCore or β-gal for 24 h then stimulated with the HCV PAMP RNA for 24 h. E) IFNβ promoter activity following 24 hours of rCore/β-gal pretreatment and 24 hour transfection of pU/UC RNA and IFNβ –firefly luciferase reporter plasmid. Units are shown as Relative Light Units (RLU) and represent the light units measured of firefly luciferase (driven by the IFNβ promoter) divided by the light units of renilla luciferase (transfection control; driven by the CMV promoter). Combined data for 3 (A, B, D & E) or 8 (C) independent experiments. P values are results of Mann-Whitney comparison of the bars indicated. *p<0.05 **p<0.01 ***p<0.001 #p≤0.0001. Mean +/− SEM.</p

rCore changes transcription factors related to IFN production.

<p>A) Top – Schematic of experimental design. Western blot of IRF-7 following 24 h of rCore/β-gal treatment (lanes 1 & 2) and 24 h of rCore/β-gal treatment followed by 24 h of IFNα (100 ng/mL) treatment (lanes 3 & 4). Arrow indicates IRF-7 band of interest. B) Schematic of experimental design. C) Western blots of IRF-7 and IRF-3 following 24 h of rCore/β-gal treatment and 24 h of HCV PAMP RNA treatment. Densitometry for IRF-7 (left) and IRF-3 (right). D) Western blots of IRF-3 and IRF-3pS396 following 24 h of rCore/β-gal treatment and 24 h of HCV PAMP RNA treatment. Densitometry for IRF-3 (left) and pIRF-3 (right). Graphs show combined densitometry data after normalization to the loading control for 3 independent experiments. Images are representative blots. P values are results of Mann-Whitney comparison of the bars indicated. **p<0.01. Mean +/− SEM.</p

Paradigm Model of HCV Core acting on pDCs.

<p>A) pDCs respond to TLR stimulation and HCV PAMP to produce IFNs Type I and IFNLs. However, in the presence of HCV core (B), there is increased STAT1 but decreased IFNs production. The decreased IFN results in decreased IRF-7, which is an ISG.</p

rCore alters JAK-STAT proteins.

<p>A) Representative histograms (left) and MFI (right) of STAT1 from 0 and 24 hours of rCore/β-gal protein exposure. B) MFI graphs of phosphoflow for STAT1 (left), STAT1pY701 (middle) and STAT1pS727 (right) after treatment with rCore/β-gal (top row), rCore/β-gal for 24 h followed by IFNα (100 ng/mL) stimulation (middle row) or rCore/β-gal for 24 h followed by pU/UC RNA stimulation (bottom row). C) Immunofluorescence (IF) showing STAT1 in rCore/β-gal pretreated cells followed by pU/UC RNA stimulation. Green – Total STAT1 Blue – nuclei. D) STAT1 and STAT1pY701 shown by Western Blot. Cells were treated for 24 hours with rCore/β-gal then stimulated with pU/UC RNA over time. Normalized densitometry shown on right. Representative blots, images and flow plots are shown. Graphs show combined data for 3 independent experiments. P values are results of Mann-Whitney comparison of the dots or bars indicated. *p<0.05 **p<0.01. Mean +/− SEM.</p