Investigation of the role of interleukin-1 family members, IL-33 and IL-36, in the pathogenesis of colon cancer

The importance of inflammation in cancer is well established, with cytokines/chemokines playing an important role in carcinogenesis. IL-33 was recently identified as the ligand for ST2. ST2 is a member of the toll-like receptor/IL-1 receptor family. Three isoforms of ST2 exist: a trans-membrane receptor (ST2L), a secreted soluble form (sST2), and a variant form (ST2V). The IL-33/ST2 pathway has been implicated in inflammatory bowel disease, a major risk factor for colon cancer. The aim of the first part of my thesis was to investigate the role of IL-33 and ST2 in colon cancer. CT26 and HT29 colon cancer cells were found to express ST2 and IL-33 in vitro, with expression increased by inflammatory mediators (LPS, TNF-α and PGE2). Stimulation with IL-33 increased the migration, but not proliferation, of colon tumour cells. Functional analyses showed that stimulation with IL-33 induced the expression of CXCL-1 by CT26 and CCL2 expression by HT29 cells. To investigate the role of ST2 in vivo, ST2 knockdown cells were generated using ST2-specific shRNA (CT26 ST2shRNA) and injected subcutaneously into BALB/C mice. Knockdown of ST2 in colon tumours resulted in enhanced tumour growth (2.3 fold increase compared to CT26 scrshRNA) in vivo. This was associated with alterations in immune cell infiltration, including an increase in macrophage infiltration. In contrast, characterisation of human colon tumours revealed that ST2L expression was increased in tumour cells relative to adjacent non-tumour cells, with no change in expression of total ST2. These results indicate that the IL-33/ST2 signalling axis may play an important role in colon carcinogenesis and merits further investigation. The role of the IL-36R, a second IL-1R family member, in colon cancer was investigated in the second part of my thesis. Characterization of human colon tumours ex vivo showed significantly increased expression of the IL-36 ligands, IL-36α and IL-36γ, compared to adjacent non-tumour tissue. In vitro colon cancer cell lines HT29 and SW480 were shown to express the IL-36R and IL-36 ligands. IL-36α and IL-36γ stimulation of HT29 cells also increased the expression of the chemokines CXCL-1, CCL2, CCL20, and IL-8. This suggests that IL-36 signalling may promote tumour-derived immune cell recruitment. This field requires further study to determine if the recruitment of immune cells by IL-36 signalling could be utilised to break tolerance against tumour antigens. This thesis has laid the basis for further studies to explore the role of IL-36 signalling in colon cancer

Wounds that heal and wounds that don't - The role of the IL-33/ST2 pathway in tissue repair and tumorigenesis

IL-33 is a member of the IL-1 family of cytokines. IL-33 is predominantly located within the nucleus of cells where it plays a role in gene regulation. Given the right combination of signals and cellular damage, stored IL-33 is released from the cell where it can interact with its receptor ST2, triggering danger-associated responses and act as a cellular "alarmin". Whilst IL-33/ST2 signalling has been shown to induce potent pro-inflammatory responses that can be detrimental in certain disease states, a dichotomous, protective role of IL-33 in promoting wound healing has also emerged in multiple tissues types. This review will explore the current literature concerning this homeostatic role of IL-33/ST2 in tissue repair and also review its role in uncontrolled wound responses as seen in both fibrosis and tumorigenesis

The Environmental Context and Function of Burnt-Mounds : New Studies of Irish Fulachtaí Fiadh

The authors acknowledge funding from The Leverhulme Trust (F/00144/AI) and assistance from a large number of individuals including; Margaret Gowen (access to sites and assistance throughout),A. Ames, H, Essex (pollen processing), S. Rouillard & R. Smith (illustrations), C. McDermott, S. Bergerbrandt, all the staff of Margaret Gowen & Co. Ltd, TVAS Ireland and CRDS. Excavation works and some post-excavation analysis was paid for my Bord Gáis and the National Roads Authority (now Transport Infrastructure Ireland). Thanks also to David Smith for access to the Maureen Girling collection and assistance with identifications.Peer reviewedPostprintPostprin

The environmental context and function of Burnt-Mounds: new studies of Irish Fulachtaí Fiadh

Burnt mounds, or fulachtaí fiadh as they are known in Ireland, are probably the most common prehistoric site type in Ireland and Britain. Typically Middle–Late Bronze Age in age (although both earlier and later examples are known), they are artefact-poor and rarely associated with settlements. The function of these sites has been much debated with the most commonly cited uses being for cooking, as steam baths or saunas, for brewing, tanning, or textile processing. A number of major infrastructural development schemes in Ireland in the years 2002–2007 revealed remarkable numbers of these mounds often associated with wood-lined troughs, many of which were extremely well-preserved. This afforded an opportunity to investigate them as landscape features using environmental techniques – specifically plant macrofossils and charcoal, pollen, beetles, and multi-element analyses. This paper summarises the results from eight sites from Ireland and compares them with burnt mound sites in Great Britain. The fulachtaí fiadh which are generally in clusters, are all groundwater-fed by springs, along floodplains and at the bases of slopes. The sites are associated with the clearance of wet woodland for fuel; most had evidence of nearby agriculture and all revealed low levels of grazing. Multi-element analysis at two sites revealed elevated heavy metal concentrations suggesting that off-site soil, ash or urine had been used in the trough. Overall the evidence suggests that the most likely function for these sites is textile production involving both cleaning and/or dyeing of wool and/or natural plant fibres and as a functionally related activity to hide cleaning and tanning. Whilst further research is clearly needed to confirm if fulachtaí fiadh are part of the ‘textile revolution’ we should also recognise their important role in the rapid deforestation of the wetter parts of primary woodland and the expansion of agriculture into marginal areas during the Irish and British Bronze Ages

IL-36 signalling enhances a pro-tumorigenic phenotype in colon cancer cells with cancer cell growth restricted by administration of the IL-36R antagonist.

The IL-36 cytokines are a recently described subset of the IL-1 family of cytokines, shown to play a role in the pathogenesis of intestinal diseases such as Inflammatory Bowel Disease (IBD). Given the link between IBD and colitis -associated cancer, as well as the involvement of other IL-1 family members in intestinal tumorigenesis, the aim of this work was to investigate whether IL-36 cytokines play a role in the pathogenesis of colon cancer. Whilst research to date has focused on the role of IL-36 family members in augmenting the immune response to induce tumour rejection, very little remains known about IL-36R signalling in tumour cells in this context. In this study we demonstrate that expression of IL-36 family member mRNA and protein are significantly increased in colorectal cancer tissue compared to adjacent non-tumour. In vitro assays showed stimulation of colon cancer cell lines with IL-36R agonists resulted in the activation of the pro-tumorigenic phenotypes of increased cellular migration, invasion and proliferation in both 2D and 3D models. In addition, the IL-36 cytokines induced strong expression of pro-inflammatory chemokines in both human and murine cell lines. Intraperitoneal injection of IL-36Ra significantly reduced tumour burden using the subcutaneous CT26 tumour model in syngeneic Balb/mice, and this was associated with a decrease in Ki-67 expression by tumour cells in the IL-36Ra- treated group relative to untreated, suggesting the inhibition of the pro-proliferative signalling of IL-36 agonists resulted in the decreased tumour size. Moreover, colon cancer cells lacking the IL-36R also showed reduced tumour growth and reduced Ki-67 expression in vivo. Taken together, this data suggests that targeting IL-36R signalling may be a useful targeted therapy for colorectal cancer patients with IL-36R+ tumour cells

Wolves in the Wolds: Late Capitalism, the English Eerie, and the Wyrd Case of ‘Old Stinker’ the Hull Werewolf

In this article, I depart from the earlier opinions of Emily Gerard, Sabine Baring-Gould, and others, who explained the disappearance of the werewolf in folklore as following the extinction of the wolf. I argue instead that British literature is distinctive in representing a history of werewolf sightings in places in Britain where there were once wolves. I draw on the idea of absence, manifestations of the English eerie, and the turbulence of England in the era of late capitalism to illuminate my analysis of the representation of contemporary werewolf sightingsPeer reviewe

An antitumorigenic role for the IL-33 receptor, ST2L, in colon cancer

Background: Despite the importance of inflammation in cancer, the role of the cytokine IL-33, and its receptor ST2, in colon cancer is unclear. The aim of this study was to investigate the role of IL-33, and its receptor isoforms (ST2 and ST2L), in colon cancer. Methods: Serum levels of IL-33 and sST2 were determined with ELISA. ST2 and IL-33 expression was detected with quantitative real-time PCR (qRT–PCR), western blotting and immunohistochemistry. ST2 expression in CT26 cells was stably suppressed using ST2-specific shRNA. Cytokine and chemokine gene expression was detected with qRT–PCR. Results: Human colon tumours showed lower expression of ST2L as compared with adjacent non-tumour tissue (P<0.01). Moreover, the higher the tumour grade, the lower the expression of ST2L (P=0.026). Colon cancer cells expressed ST2 and IL-33 in vitro. Functional analyses showed that stimulation of tumour cells with IL-33 induced the expression of chemokine (C–C motif) ligand 2 (CCL2). Knockdown of ST2 in murine colon cancer cells resulted in enhanced tumour growth (P<0.05) in BALB/c mice in vivo. This was associated with a decrease in macrophage infiltration, with IL-33-induced macrophage recruitment reduced by antagonising CCL2 in vitro. Conclusion: The IL-33/ST2 signalling axis may have a protective role in colon carcinogenesis

Transactional sex among young women in rural South Africa: prevalence, mediators and association with HIV infection.

INTRODUCTION: Young adolescent women in sub-Saharan Africa are three to four times more likely to be HIV-positive than boys or men. One of the relationship dynamics that is likely to be associated with young women's increased vulnerability to HIV is transactional sex. There are a range of HIV-related risk behaviours that may drive this vulnerability. However, to date, limited epidemiological data exist on the role of transactional sex in increasing HIV acquisition, especially among young women in sub-Saharan Africa. Our paper presents data on the prevalence of self-reported engagement in transactional sex and explores whether transactional sex is associated with increased risk of HIV infection among a cohort of young, rural, sexually active South African women. We also explore whether this relationship is mediated through certain HIV-related risk behaviours. METHODS: We analyzed baseline data from a phase III trial of conditional cash transfers for HIV prevention of 693 sexually active, school-going young women aged 13-20 years in rural South Africa. We examined the association between young women's engagement in transactional sex and HIV infection. Transactional sex is defined as a non-commercial, non-marital sexual relationship whereby sex is exchanged for money and/or gifts. We explored whether this relationship is mediated by certain HIV-related risk behaviours. We used logistic and multinomial regression and report unadjusted and adjusted odds ratios with 95% CI. RESULTS: Overall, 14% (n=97) of sexually active young women reported engaging in transactional sex. Engagement in transactional sex was associated with an increased risk of being HIV-positive (aOR: 2.5, CI: 95% 1.19-5.25, p=0.01). The effect size of this association remained nearly unchanged when adjusted for certain other dimensions of HIV risk that might help explain the underlying pathways for this relationship. CONCLUSIONS: This study provides quantitative support demonstrating that transactional sex is associated with HIV infection in young women. Even though the specific variables tested do not mediate the relationship, a potential explanation for this association may be that the men with whom young women are having sex belong to networks of sexually connected individuals who are at a "high risk" for HIV infection. The results highlight the importance of structural intervention approaches that can alter the context of young women's HIV risk

Human aquaporins: regulators of transcellular water flow

Background: Emerging evidence supports the view that (AQP) aquaporin water channels are regulators of transcellular water flow. Consistentwith their expression in most tissues, AQPs are associatedwith diverse physiological and pathophysiological processes. Scope of review: AQP knockout studies suggest that the regulatory role of AQPs, rather than their action as passive channels, is their critical function. Transport through all AQPs occurs by a common passive mechanism, but their regulation and cellular distribution varies significantly depending on cell and tissue type; the role of AQPs in cell volumeregulation (CVR) is particularly notable. This reviewexamines the regulatory role of AQPs in transcellular water flow, especially in CVR.We focus on key systems of the human body, encompassing processes as diverse as urine concentration in the kidney to clearance of brain oedema. Major conclusions: AQPs are crucial for the regulation of water homeostasis, providing selective pores for the rapidmovement ofwater across diverse cellmembranes and playing regulatory roles in CVR. Gatingmechanisms have been proposed for human AQPs, but have only been reported for plant andmicrobial AQPs. Consequently, it is likely that the distribution and abundance of AQPs in a particular membrane is the determinant of membrane water permeability and a regulator of transcellular water flow. General significance: Elucidating the mechanisms that regulate transcellular water flow will improve our understanding of the human body in health and disease. The central role of specific AQPs in regulating water homeostasis will provide routes to a range of novel therapies. This article is part of a Special Issue entitled Aquaporins