NIHRDataInsights/HRCSData huggingface dataset:
Title: CRUK/02/010: OE05: A randomised controlled trial comparing standard chemotherapy followed by resection versus ECX chemotherapy followed by resection in patients with resectable adenocarcinoma of the oesophagus.
Funder: Cancer Research UK

Abstract: Background: Cancer of the oesophagus causes over 7000 deaths in the UK each year . All potentially curative treatments involve surgery, but when it is used alone only around 20% of patients are alive after 3 years. The MRC trial (OE02) showed survival was significantly prolonged (HR 0.84 95% CI 0.72-0.98 p=0.03) by giving two cycles of cisplatin and 5-fluorouracil (CF) chemotherapy pre-operatively. Aims: The primary objective is to evaluate whether pre-operative prolonged ECX (epirubicin, cisplatin and capecitabine) chemotherapy improves overall survival compared to the standard shorter pre-operative OEO2 CF chemotherapy in patients undergoing oesophagectomy for adenocarcinoma of the oesophagus and gastro-oesophageal junction. Trial design: OE05 is an open, randomised-controlled, phase III trial with a minimum target accrual of 842 patients. Participants are randomised to receive either 2 cycles of CF chemotherapy, or 4 cycles of ECX chemotherapy prior to resection (randomisation ratio 1:1). The primary outcome measure is overall survival (OS), secondary outcome measures include disease-free survival (DFS), treatment morbidity and quality of life (QL).


Research activity codes: 6.4 SURGERY;6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Mechanisms of tumour invasion in Drosophila models of metastasis
Funder: Cancer Research UK

Abstract: The fruitfly Drosophila has been increasingly successfully used to study the biological processes underlying cancer. Several tumour models have been developed, all sharing over-proliferative and invasive/metastatic cell behaviour, which eventually kills the animal. The genetic advantages of using Drosophila, together with the conservation of fundamental biological mechanisms between flies and mammals, makes Drosophila an excellent model system. We plan to capitalise on this to analyse the effect of invasion of wild type tissue by tumour cells. The particular goal of this study will be to investigate the genetic requirements in the target tissue for invasion to occur. This is difficult to study in mammalian systems with less amenable genetic technologies, yet is an important hallmark of cancer morbidity and mortality: substantial damage is caused by the destruction of healthy tissue once the primary tumour spreads. We will use two well established fly tumour models. The first combines a loss of function of the tumour suppressor gene, scribbled, a component of the apical-basal polarity machinery, with an activated form of the oncogene ras. The second system is a recently developed model of human multiple endocrine neoplasia type II (MEN2). The use of two distinct cancer models will allow us to compare the biology of their invasion, showing which aspects are common between different tumours, and which are specific to each. We have recently established the first ex-vivo imaging system to study post embryonic Drosophila development, and in the proposed work will extend this to allow real-time imaging of tumour invasion. A dual genetic system will be developed to produce tumours in a discrete territory, while simultaneously genetically modifying the invaded tissue. This will allow the analysis of candidate genes that are predicted to be involved in response to tumour invasion. In addition, we will perform a whole genome RNAi screen to identify a comprehensive list of genes needed in the target tissue for successful tumour invasion. The use of the combined genomic/imaging approach will give us a comprehensive picture of how the tumours spread and invade the wild type tissue, and of the response generated by the invasion. Since the underlying cell biology of Drosophila and humans is similar, this should provide potential leads for novel therapeutic strategies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Establishment of sister chromatid cohesion by cohesin
Funder: Cancer Research UK

Abstract: By holding sister chromatids together from S phase until the onset of anaphase, a proteinaceous ring complex called cohesin has as key a role in chromosome segregation as the microtubules that actually move chromosomes to the metaphase plate and chromatids to opposite poles of the cell prior to its division. The mechanism by which cohesion is established and maintained is poorly understood. Due to its unique advantages for genetical, physiological, cytological, and biochemical analyses, we propose to study cohesin's role in the budding yeast S.cerevisiae. Because no work on cohesin makes any sense without understanding the physical nature of sister chromatid cohesion, an important early priority will be to test the generality of the ring model, according to which sister DNAs are entrapped inside individual cohesin rings. To do this, we will develop alternative methods of cross-linking inter-subunit interactions and test whether this entraps, within chemically circularized cohesin rings, sister DNAs from minichromosomes up to 25 kb in size and whether such entrapment is a property of cohesion established by arm as well as centromere sequences. Because cohesin associated with an individual chromatid is a possible precursor to cohesion itself, we will use cross-linking to investigate whether this association is also mediated by entrapment within cohesin rings. We will investigate the mechanism of chromatin fibre entrapment, in particular the role of chromosomal proteins at centromeres and at highly transcribed genes, and the consequences of entrapment, namely whether rings subsequently diffuse along chromatin fibres. Because deep mechanistic understanding will depend on knowing the structures of cohesin subunits, we will devote considerable effort into determining the structures of cohesin's Scc3 and Pds5 subunits as well as that of its Scc2/4 loading complex. We will use live imaging of GFP-tagged cohesin subunits to study changes in cohesin's turnover on chromatin before and after DNA replication, to study the role of acetylation by Eco1 in this process, and to determine whether cohesin rings associated with unreplicated chromatids are converted in situ to rings that entrap sisters after DNA replication. Lastly, we will investigate the molecular mechanisms that regulate acetylation during S phase and de-acetylation during anaphase of cohesin's Smc3 subunit, the role of Smc3 acetylation in maintaining cohesion in post-replicative cells, and the importance of de-acetylation for reforming a new round of cohesion.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Contribution of aberrant lipid signalling to the tumour micro-environment in Hodgkin's lymphoma
Funder: Cancer Research UK

Abstract: BACKGROUND: In Hodgkin’s lymphoma, the malignant HRS cells are surrounded by a prominent tumour micro-environment which has many features in common with other solid cancers. This micro-environment can suppress anti-tumour immune responses as well as directly support tumour cell growth. Two phospholipids, sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) have been implicated in the development and maintenance of the tumour microenvironment. Previously, we have shown that both lipids are aberrantly regulated in HRS cells. RELEVANCE TO CANCER: Tumour cells have been shown to contribute to the development and maintenance of the tumour micro-environment. Disrupting such tumour cell-stromal interactions is now considered a potentially important therapeutic approach. In this proposal we will study how S1P and LPA contribute to the tumour microenvironment in HL, and determine the extent to which these effects can be reversed using anti-lipid therapies. SPECIFIC AIMS The objectives of this study are to: - Study the influence of S1P and LPA on the phenotype, chemotaxis, and function of lymphoid stromal cells. - Investigate in a 3-dimensional co-culture system, the influence of HRS cell-derived factors on the phenotype of lymphoid stromal cells, and to determine if these changes can be reversed by lipid signalling antagonists. - By implantation into NOD/Shi-scid IL2rgamma null mice, establish a system that allows the HL tumour/stroma micro-environment to be maintained and analysed in vivo. Use this system to explore the impact on HRS cells and on the tumour micro-environment of lipid signalling antagonists. METHODS We will study the influence of S1P and LPA on the proliferation, apoptosis, and chemotaxis, of lymphoid stromal cells (T lymphocytes, macrophages, fibroblasts) isolated from human tonsils by microbead technology and MACS. We will determine which of the S1P and LPA receptors mediate these effects using siRNA, and receptor antagonists/blocking antibodies. We will use microarrays to explore the mechanisms responsible. We will use a 3-dimensional co-culture system to study the proliferation, survival and gene expression of the lymphoid stromal cells cultured either alone or in the presence of HL supernatant and following addition of S1P and LPA blocking antibodies. We will use NOD/Shi-scid IL2rgamma null (NOG) to establish an in vivo system in which tumour cell/stromal cells interactions can be maintained. We will use this system to explore the influence of S1P and LPA signalling antagonists on these interactions.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: High-throughput in vivo screens for the genetic determinants of genomic stability and cancer susceptibility
Funder: Cancer Research UK

Abstract: The vast majority of cancer cells become genomically unstable following the acquisition of mutations in DNA repair genes. We will screen for genes that regulate genomic stability by phenotyping mice for micronucleus induction. Micronuclei are fragments of extranuclear chromatin that have failed to be incorporated into daughter nuclei during cell division. The frequency of micronucleated cells is a useful biomarker of cancer risk. We have established a high-throughput flow cytometry-based method of micronucleus scoring that greatly increases the speed and sensitivity of detection over microscope-based methods. Our pilot studies have identified a number of genes that contribute to genome stability. These data strongly suggest that patients who have lost these genes may be predisposed to cancer and should participate in cancer surveillance programs With this application we wish to screen 1. up to 500 knockouts per year for increased levels of genomic instability as a hallmark for cancer predisposition and 2. 2000 genetically diverse mouse stocks for genes and interactions between genes (epistasis) that regulate genomic stability.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Structural basis of membrane trafficking and signalling interactions of proteins implicated in cancer progression
Funder: Cancer Research UK

Abstract: Our goal is to understand how signalling proteins are recruited to and traffic membranes, and to elucidate the effects of mutations linked to cancer on these interactions. We are particularly interested in how phosphoinositides and organelle membranes are specifically targeted by protein domains. In so doing we have discovered how proteins tubulate bilayers and direct traffic between organelles and the cell surface. Here we focus on the pleckstrin homology (PH) domain, the most widespread type of membrane-targeting module. Although known to play central roles in regulating signal transduction and cancer progression, the scarcity of quality structural and binding information on the bilayer complexes and protein assemblies has limited progress, and new experimental tools are needed. We hypothesize that membranes are generally engaged through concerted steps including (1) non-specific and transient electrostatic approach, (2) specific phospholipid recognition, (3) stable and oriented hydrophobic bilayer insertion, and (4) formation of protein multimers and complexes, which responsible for bending, tubulating and vesiculating the bilayer during trafficking events. This process is critical for cancer biology, being lined by transforming mutations and dictating the sphere of activity of oncoproteins and wild type forms We will map the networks of protein-lipid and protein-protein contacts that underlie these encounters, and that induce changes in protein and bilayer structure. The techniques used for structural analysis of will be performed at the UK’s largest nuclear magnetic resonance (NMR) facility installed within the Birmingham Cancer Research UK Centre; small angle X-ray scattering data will also be collected from synchrotron sources. We will develop and apply novel research tools for producing peripheral membrane proteins, predicting membrane interaction sites and solving lipid micelle complexed structures, thus overcoming major technical barriers within the field, providing general applicability to virtually any membrane-associated protein. The outcomes will include solution structures of multidomain proteins including FAPP1 and AKAP13 bound to lipid micelles and protein partners in physiological solutions. The lipid binding and membrane tubulation properties of full length wild type and relevant cancer mutant forms will be carried out by surface plasmon resonance and confocal microscopy. Novel biological ligands and allosteric sites and druggable pockets will be investigated by thermal shift, NMR and phage display library screening. The results will be generalized and extension to other proteins involved in cancer progression. Our endgoal is to develop a more predictive understanding of the molecular behaviour of oncoproteins in cellular environments, and to identify and develop membrane interaction sites as tractable sites for drug design.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: JMJD2B and MLL2 in androgen receptor signalling in prostate cancer
Funder: Cancer Research UK

Abstract: The Androgen Receptor (AR) is important in prostate cancer (PC) development and progression rendering it a significant drug target. Current therapies have short term effectiveness highlighting the importance of identifying novel therapeutic targets. The AR, a member of the steroid hormone receptor family, can interact with multiple co-regulatory proteins to modulate AR transcriptional output, providing alternative therapeutic targets. Although we have previously identified the histone methyltransferase (HMT) SET7/9 as an AR co-regulator protein, the role of other HMT and histone demethylases (HDM) in receptor regulation remains unclear. We have since identified the HDM, JMJD2B, and the HMT, MLL2, as being involved in AR regulation. Knockdown of either JMJD2B or MLL2 reduced expression of androgen regulated genes PSA, TMPRSS2 and KLK2 and decreases proliferation in cell line models. MLL2 resides in a complex with other proteins similar to the methyltransferase complex, COMPASS, in yeast. H2B ubiquitination is required for the complex to methylate H3K4 and activate transcription. Our initial data clearly indicates that MLL2 plays a role in the regulation of AR mediated transcription but how this is achieved is currently unknown. JMJD2B knockdown reduced migration and AR mRNA and protein levels in both castrate sensitive and castrate resistant cell line models and JMJD2B itself is up-regulated by androgens and can increase AR transcriptional activity in a demethylase-dependent manner. This suggests a close relationship between AR and JMJD2B. To further determine the roles of MLL2 and JMJD2B in androgen regulated gene expression and PC we will use the following strategies: - 1. Investigate the interplay between histone methylation, acetylation and ubiquitination using AR regulated transcription as a model. Question whether MLL2 and JMJD2B function in the same chain of events leading to transcription initiation. - 2. Assess the role of JMJD2B in AR regulation. Chromatin immunoprecipitation (ChIP)-sequencing will determine JMJD2B chromatin binding sites, identifying JMJD2B regulated genes to determine which cellular pathways may be affected. We will study the AR promoter to determine whether JMJD2B plays a role in transcriptional activation. - 3. Characterise phenotypic effects of JMJD2B. Using clinical PC specimens JMJD2B expression will be correlated with clinical parameters. In vitro, the role of JMJD2B in the cell cycle, proliferation, migration and invasion will be investigated.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Effect and significance of Notch/androgen signalling interactions in prostate cancer
Funder: Cancer Research UK

Abstract: Growth and differentiation of the prostate is driven by androgens, effects of which are mediated by the androgen receptor (AR). On binding androgen the AR translocates from cytoplasm to nucleus, where it binds specific DNA sequences in regulatory regions of target genes and recruits proteins including coactivators and corepressors, to form complexes that modulate transcription by altering chromatin accessibility. Initially, prostate tumour growth is androgen-dependent and treatment involves androgen ablation and antiandrogens. Most patients respond well but after a median 2 years, tumours progress to the aggressive “androgen independent” stage of the disease. This has been attributed to mutation or amplification of the AR gene and altered levels or distribution of coactivator and corepressor proteins. At this stage patients have a median survival of 12-18 months, thus it is important to understand the mechanisms that regulate AR activity and drive cancer progression. Notch signalling regulates stem cell renewal, cell fate and differentiation and its deregulation has been implicated in prostate cancer. HEY proteins (HEY1, HEY2, HEYL) are Notch target/effector proteins and transcriptional repressors. We found that all 3 HEY proteins interact with AR and inhibit AR activity. Furthermore, it appears that HEY1 and HEYL are excluded from the nucleus of malignant prostate epithelial cells in human tumours, but not in benign tissues. Cytoplasmic localisation alone would prevent AR repression but we have also found it may be result in HEY proteins increasing AR activity. We hypothesise that HEY proteins sequester additional repressors in the cytoplasm, alleviating nuclear repression thus increasing AR activity. We propose that this may promote progression of prostate cancer to the hormone-refractory stage, and may be a feasible target to halt or even reverse this progression. We will use cancer cell lines expressing wild-type and mutant HEY proteins to characterise interactions between Notch and androgen signalling in prostate, and the consequences of dysregulation of this interaction, at the proteomic and genomic level. We will identify HEY-interacting proteins with functional roles in prostate and ultimately prostate cancer progression. We will also identify genes and pathways co-regulated by HEY proteins and AR. Finally, we will investigate HEY localisation in models of prostate cancer and also progression of human tumours carrying wild-type and mutant HEY proteins. This will demonstrate whether altered HEY localisation, as observed in human tumours, can drive prostate tumour progression.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Understanding the cellular regulation of the NHEJ double-strand break repair pathway
Funder: Cancer Research UK

Abstract: DNA double-strand breaks (DSBs) are one of the most lethal forms of damage and their repair is crucial for the maintenance of genomic integrity. Incorrect DSB repair plays an important role in the generation of cancer and this process is a major cause of genome instability. In particular, chromosomal rearrangements, such as translocation, can lead to the activation of oncogenes. Components of the DSB repair pathways also provide excellent targets for the design of chemo- and radio-therpeutic treatments. Homologous recombination (HR) accurately repairs DSBs but requires the presence of a homologous sequence. Non-homologous end-joining (NHEJ) does not rely on a homologous chomatid and is a potentially mutagenic mechanism. The two major pathways are deployed in a cell-cycle specific manner. In S/G2 phase, HR utilizes the homologous chromosome as a template for repair; in contrast, sister chromatids are not available in G0/G1, where NHEJ predominates. HR and NHEJ are regulated independently, since inactivation of one does not alter levels of the other. Though several HR factors have been shown to be targets of cell-cycle regulation, no such targets have been identified for NHEJ. Since DSB repair is abnormal in many cancer cells, a better understanding of DSB repair regulation is key to optimising therapeutic strategies. We aim to elucidate cell-cycle specific regulation of NHEJ in eukaryotic cells.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: CRUK/10/049: 6MP BRCA: Phase II Clinical Trial Of 6-Mercaptopurine and low-dose Methotrexate in Patients with BRCA Defective Tumours
Funder: Cancer Research UK

Abstract: Background: Ovarian and breast cancer are major causes of death of women across the World. BRCA1 and BRCA2 genes are critical in homologous recombination (HR) DNA repair and have been implicated in familial breast and ovarian cancer tumorigenesis. Tumour cells with these mutations demonstrate increased sensitivity to cisplatin chemotherapy and poly(ADP-ribose) polymerase (PARP) inhibitors and these are presently in clinical trials. Interestingly, Helleday et al have recently shown that 6 thioguanine (6TG) selectively kills BRCA2-defective cells as effectively as the PARP inhibitor, AGO14699. Furthermore even after BRCA2-defective cells have acquired resistance to a PARP inhibitor or cisplatin, they retain sensitivity to 6TG. Aims and objectives: To determine the response rates and toxicity of 6MP with low dose methotrexate in patients with breast or ovarian cancer who are known to have a BRCA mutation. Methods: Multicentre Phase II study recruiting 63 patients with relapsed advanced breast or known to have a BRCA mutation. Patients will receive daily 6MP (75 mg/m2) and weekly low dose methotrexate (20mg/m2) until disease progression. How results will be used: If the efficacy criteria are met as defined in the protocol, then we anticipate moving straight on to a Phase III randomised controlled trial.


Research activity codes: 6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS