Students and staff working in partnership: experiences from a collaborative writing group

Higher education institutions are striving to enhance student engagement in learning (Carini et al, 2006). Increasing the degree of student ownership of the learning process and offering an authentic situated learning experience (Brown et al, 1989) are possible ways to enhance student engagement. In response to this, participants on a postgraduate programme in Professional Education at Queen Margaret University (QMU), Edinburgh, were invited to set up a writing group in partnership with a member of staff from the programme team. Participants on this course were either lecturers at QMU, lecturers at other higher education institutions or health professionals with an education remit. All participants were under differing degrees of pressure to publish written work related to their practice and only the member of staff from the programme team had published previously. Many of the participants were not confident in their ability to produce writing for publication (Dixon 2001). This paper outlines the experiences from this collaborative writing group in which members of the group wrote an article for publication about their perceptions of being involved in an action research project. An outline is given of the aims of the writing group, the writing approach adopted, the group processes involved and the outcomes from the group. This work offers insights into how partnership working between ‘students’ and ‘academics’ as part of a course, can enhance student engagement in learning and develop their confidence to write and publish

The role of gamma delta T lymphocytes in breast cancer: a review

Gammadelta T (γδT) lymphocytes have provoked interest in oncology, particularly as regards their potential use in immunotherapy, because of their unique ability to recognise antigens without a requirement for major histocompatibility complex antigen presentation, and to quickly activate an anti-tumour response. However, work in some cancers has suggested that they also have pro-tumourigenic activity. Their role in breast cancer is unclear. This review outlines the evidence to date in in vitro studies, in vivo mouse models and in human studies regarding the role of γδT lymphocytes in breast cancer. We describe the seemingly opposing roles of the predominantly circulating Vγ9Vδ2 subtype, which can suppress tumour growth through direct cytotoxicity, induction of apoptosis and inhibition of angiogenesis, and the predominantly tumour-infiltrating γδ1 subtype which can promote tumour growth and spread through immunosuppressant effects. We summarise the evidence in breast cancer for the mechanisms of action of γδT lymphocytes and describe how factors in the tumour microenvironment may affect their function, polarising them towards a pro-tumourigenic, immune-suppressing role. We also describe the experience to date of γδT lymphocytes in immunotherapy for breast cancer and suggest the direction of work going forward, particularly as regards different breast cancer subtypes

Immunotherapy: enhancement the efficacy of this promising therapeutic in multiple cancers

Cancer treatments often reach a refractory period leading to treatment failure and patients developing disease recurrence. This can be due to tumour cells escaping the immune response and creating an immunosuppressive microenvironment enhancing cancer progression. Immunotherapy has become a promising tool for cancer treatment as it restores the anti-tumour response of the patient’s immune system. Immune checkpoint inhibitors are the most widely studied immunotherapies worldwide and are now approved for multiple cancers. However, CAR-T cell therapy has also shown promise by targeting T-lymphocytes that are genetically modified ex vivo to expressed chimeric antigen receptors and this is now approved to treat some haematological cancers. Although immunotherapy has shown successful treatment outcomes in multiple cancers, some patients do not respond to this treatment. Therefore, approaches to enhance the efficacy of immunotherapies are likely to be the key to improving their effectiveness. Therefore, combination therapies of checkpoint inhibitors +/- chemotherapy are at the forefront of current research. Furthermore, biomarkers that predict treatment response are now beginning to emerge. Additionally, utilizing nanoparticles as a new-targeted drug delivery system to enhance CAR-T cell therapy may enhance the efficacy of the cells when re-infused within the patient. Even if efficacy is enhanced, severe immune-related adverse events (irAEs) occur that are life threatening and could lead to therapy being stopped. Therefore, predictive biomarkers for toxicity are also needed to improve both the patient’s quality of life and treatment outcomes. This review will look at the current immunotherapies in clinical trials and discuss how to enhance their efficacy

Loss of signal transducer and activator of transcription 1 is associated with prostate cancer recurrence

STAT1 loss has previously been implicated in cell line studies to modify prostate cancer cell growth and survival, however the clinical significance of this has not previously been established. This study investigated if STAT1 loss was associated with patient outcome measures and the phenotypic consequence of STAT1 silencing. STAT1 expression was assessed in two patient cohorts with localised (n = 78) and advanced prostate cancer at initial diagnosis (n = 39) by immunohistochemistry (IHC). Impact of STAT1 silencing on prostate cancer cells lines was assessed using Cell Death detection ELISA, TLDA gene signature apoptosis arrays, WST-1 assay, xCELLigence system, clonogenic assay, and wound healing assay. In the localised patient cohort, low expression of STAT1 was associated with shorter time to disease recurrence (3.8 vs 7.3 years, P = 0.02) and disease specific survival (6.6 vs 9.3 years, P = 0.05). In the advanced patient cohort, low expression was associated with shorter time to disease recurrence (2.0 vs 3.9 years, P = 0.001). When STAT1 was silenced in PC3 cells (AR negative) and LNCaP cells (AR positive) silencing did not influence levels of apoptosis in either cell line and had little effect on cell viability in the LNCaP cells. In contrast, STAT1 silencing in the PC3 cells resulted in a pronounced increase in cell viability (WST-1 assay: mock silenced vs STAT1 silenced, P < 0.001), clonagenicity (clonogenic assay: mock silenced vs STAT1 silenced, P < 0.001), and migration (wound healing: mock silenced vs STAT1 silenced, P < 0.001). In conclusion, loss of STAT1 may promote prostate cancer recurrence in AR negative patients via increasing cell viability

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Phosphorylation of androgen receptors at serine 515 is a potential prognostic marker for triple negative breast cancer

1.7 million cases of breast cancer are diagnosed every year with 522,000 deaths. Molecular classifications of breast cancer have resulted in improved treatments. However, treatments for triple negative breast cancer (TNBC) are lacking. Analysis of molecular targets for TNBC is a priority. One potential candidate is androgen receptor (AR) phosphorylation. This study assessed the role of AR phosphorylation at ser81/ser515 and their two upstream effectors, cyclin-dependent kinase 1 (pCDK1) and extracellular-regulated kinase 1/2 (pERK1/2) in 332 ductal breast cancer patients by immunohistochemistry. pERK1/2 combined with AR-515 associated with improved cancer-specific survival (CSS, p = 0.038), decreased size (p = 0.001), invasive grade (p < 0.001), necrosis (p = 0.003), b-lymphocytes (p = 0.020), molecular subtype (p < 0.001) and estrogen receptor (ER)/progesterone receptor (PR)-status (p < 0.001). The cohort was therefore stratified into ER+ve and ER-ve patients. In ER+ve tumours, pERK1/2 combined with AR-515 associated with improved CSS (p = 0.038), smaller size (p = 0.004), invasive grade (p = 0.001), decreased b-lymphocytes (p = 0.013) and increased plasma cells (p = 0.048). In contrast, in TNBC patients, phosphorylation of AR-515 associated with poorer CSS (p = 0.007). pERK1/2 combined with AR-515 associated with decreased inflammation (p = 0.003), increased tumour stroma (p = 0.003) and tumour budding (p = 0.011), with trends towards decrease CSS (p = 0.065) and macrophage levels (p = 0.093). In Conclusions, AR-515 may be an important regulator of inflammation in breast cancer potential via ERK1/2 phosphorylation. AR-515 is a potential prognostic marker and therapeutic target for TNBC

The prognostic role of the non-canonical nuclear factor-kappa B pathway in renal cell carcinoma patients

Background: In the United Kingdom, 8,000 cases of renal cancer are diagnosed each year, with a 5-year survival rate of 50%. Treatment options are limited; a potential therapeutic target is the non-canonical nuclear factor-kappa B (NF-κB) pathway. This pathway plays a role in multiple oncogenic processes in solid tumors. The aim of this study was to investigate the non-canonical nuclear factor pathway in renal cell carcinoma (RCC). Materials and Methods: NIK, IKKα, and RelB were investigated via immunohistochemistry in a cohort of 192 patients with clear cell renal cancer. Results: High cytoplasmic NIK was associated with poorer cancer-specific survival (p = 0.006) and 10-year survival stratified from 85% (low) to 65% (high, p = 0.005). Similarly, high cytoplasmic RelB was associated with poorer cancer-specific survival (p = 0.041) and 10-year survival stratified from 88% (low) to 73% (high, p = 0.030). When clinicopathological characteristics were assessed, cytoplasmic NIK was associated with survival (p = 0.014), whereas cytoplasmic RelB was associated with increased tumor grade (p = 0.020) and decreased inflammation (p = 0.019). Upon multivariate analysis, it was found that cytoplasmic NIK was independently associated with cancerspecific survival (p = 0.009). Conclusions: The non-canonical NF-κB pathway is associated with poorer cancer-specific survival in RCC patients, making it a viable target for therapeutic intervention. Furthermore, cytoplasmic NIK is a potential prognostic biomarker for this disease

Functional analyses of the role of kisspeptins and their receptor, gpr-54 in the biology of reproductive tissues

GnRH neurons represent the final common pathway for the regulation of the reproductive axis and they are modulated by multiple signals. It has recently been shown that a potent effector of GnRH neuron function is an afferent network of kisspeptin-producing neurons. Kisspeptin released from these neurons acts upon a specific receptor (gpr-54) expressed on GnRH neurons, and increases the secretion of GnRH from the hypothalamus. The kisspeptin system has since been implicated as a downstream mediator for regulation of the Hypothalamic-Pituitary-Gonadal (HPG) axis by steroid hormones, metabolic signals and photoperiod, potentially placing it at the centre of reproductive physiology. However, the supporting evidence to date has been indirect, relying on interpretation of changes in mRNA levels and immuno-histochemical staining to infer the actions of kisspeptin upon the central control of reproduction. The detailed mechanisms of kisspeptin action are yet to be fully elucidated. The research within this thesis elucidates the effect of kisspeptin on the HPG axis via the development of kisspeptin-10 (kp-10) analogues with antagonistic properties. Functionally important residues within the peptide were delineated. Structure-activity studies of kp-10 analogues indicated that residues Asn2, Trp3, Phe6, Arg9 and Phe10 interact with gpr-54 to facilitate receptor binding. Two other residues, Tyr1 and Leu8 were shown to be critical for receptor activation by kisspeptin. Four synthetic peptide antagonists were selected according to a consensus sequence for good antagonism: X1-N-W-N-X5-F-G-X8-R-F-NH2 where X1 = D-Ala or D-Tyr, X5 = Gly or D-Ser and X8 = D-Trp or D-Leu. One of the antagonists, peptide 234, was used in in vivo studies, where it inhibited the amplitude of GnRH and LH pulses without affecting basal secretion of GnRH or LH. These results indicate for the first time that basal and pulsatile secretion of these factors is regulated by separate pathways. Use of the antagonist also demonstrated the direct involvement of endogenous kisspeptin in steroid hormone negative feedback, positive regulation of the pre-ovulatory LH surge and in regulating the onset of puberty in rodents, as had been suggested via indirect methods. Although a major role of the kisspeptin system is in the regulation of the HPG axis, the system may also be important in the inhibition of cancer cell metastasis and in placental development (trophoblast cell invasion) but little is known about the mechanisms involving kisspeptin in these processes. This thesis describes novel signalling mechanisms for the regulation cell migration by kisspeptin, involving the MAPK and GSK3β signalling pathways. Using a stably transfected CHO cell line, kisspeptin-gpr-54 signalling can activate all members of the MAPK pathway, the β- catenin/GSK3β pathway, NFκB and FAK. These factors are involved in inhibiting the migration of these cells via an ERK1/2-p90rsk-GSK3β-β catenin pathway to potentially up- regulate formation of adherens junctions at the plasma membrane. This pathway was also shown to be involved in the inhibition of migration within an immortalised human first trimester placental trophoblast cell line and in human umbilical vein endothelial cells. Some of these pathways were also active within a mouse GnRH neuronal cell line, where ERK1/2, NFκB and GSK3β were activated by kisspeptin with no effect on migration. However, the role of these pathways in the GnRH neuronal cells requires further investigation. In summary, the research presented within this thesis defines receptor-binding and activating residues within kisspeptin-10, which should enable more details of ligand-receptor binding interactions to be fully elucidated. Novel gpr-54 antagonists have been identified and used in in vivo studies. The thesis demonstrates the direct involvement of endogenous kisspeptin in the regulation of GnRH/LH secretion at the onset of puberty and throughout the reproductive cycle in mature animals. The antagonists developed within this thesis represent useful tools to further delineate mechanisms of kisspeptin action within the HPG axis and peripheral tissues. Other findings describe kisspeptin signalling mechanisms for the inhibition of cell migration, potentially important in a variety of normal and pathological processes, including for the first time a description of the regulation of GSK3β and β-catenin signalling factors by kisspeptin and gpr-54

Nuclear expression of Lyn, a Src family kinase member, is associated with poor prognosis in renal cancer patients

Background: 8000 cases of renal cancer are diagnosed each year in the UK, with a five-year survival rate of 50 %. Treatment options are limited; a potential therapeutic target is the Src family kinases (SFKs). SFKs have roles in multiple oncogenic processes and promote metastases in solid tumours. The aim of this study was to investigate SFKs as potential therapeutic targets for clear cell renal cell carcinoma (ccRCC). Methods: SFKs expression was assessed in a tissue microarray consisting of 192 ccRCC patients with full clinical follow-up. SFK inhibitors, dasatinib and saracatinib, were assessed in early ccRCC cell lines, 786-O and 769-P and a metastatic ccRCC cell line, ACHN (± Src) for effects on protein expression, apoptosis, proliferation and wound healing. Results: High nuclear expression of Lyn and the downstream marker of activation, paxillin, were associated with decreased patient survival. Conversely, high cytoplasmic expression of other SFK members and downstream marker of activation, focal adhesion kinase (FAK) were associated with increased patient survival. Treatment of non-metastatic 786-O and 769-P cells with dasatinib, dose dependently reduced SFK activation, shown via SFK (Y419) and FAK (Y861) phosphorylation, with no effect in metastatic ACHN cells. Dasatinib also increased apoptosis, while decreasing proliferation and migration in 786-O and 769-P cell lines, both in the presence and absence of Src protein. Conclusions: Our data suggests that nuclear Lyn is a potential therapeutic target for ccRCC and dasatinib affects cellular functions associated with cancer progression via a Src kinase independent mechanism

Relationship between tumour PTEN/Akt/COX-2 expression, inflammatory response and survival in patients with colorectal cancer

In patients with colorectal cancer (CRC), local and systemic inflammatory responses have been extensively reported to associate with cancer survival. However, the specific signalling pathways responsible for inflammatory responses are not clear. The PTEN/Akt pathway is a plausible candidate as it may play a role in mediating inflammation via COX-2, and has been associated with cancer progression. This study therefore examined the relationship between tumour PTEN/Akt/COX-2 expression, inflammatory responses and survival in CRC patients using a tissue microarray. In 201 CRC patients, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (12.0yrs v 7.3yrs, P=0.032), poorer differentiation (P=0.032), venous invasion (P=0.008) and peritoneal involvement (P=0.004). Patients were stratified for peri-nuclear expression of COX-2 to examine associations with inflammatory responses. In patients with absent peri-nuclear COX-2 expression, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (11.9yrs v 5.4yrs, P=0.001), poorer differentiation (P=0.018), venous invasion (P=0.003) and peritoneal involvement (P=0.001). However, no associations were seen with either the local or systemic inflammatory responses. In CRC patients, tumour-specific PTEN/Akt pathway activation was significantly associated with poorer CSS, particularly when peri-nuclear COX-2 expression was absent. However, activation of the PTEN/Akt pathway appears not to be responsible for the regulation of inflammatory responses