NIHRDataInsights/HRCSData huggingface dataset:
Title: The mechanisms and regulation of the APC/C ubiquitin ligase system
Funder: Cancer Research UK

Abstract: Background_x000D_ The anaphase-promoting complex/cyclosome (APC/C) is an essential multi-subunit ubiquitin ligase complex and serves as an important regulator of the cell cycle, ensuring genome integrity. Deregulation of the APC/C leads to aneuploidy, thus the APC/C has to be tightly controlled, which is achieved by a number of regulatory networks. In addition, the APC/C co-activators, Fizzy/Cdc20 or Fizzy-related/Cdh1 play a role in substrate recognition and the catalytic activation of the APC/C. Despite extensive genetic and biochemical studies, little is known about the precise mechanism of APC/C substrate recognition and ubiquitylation of specific substrates or how APC/C activity is controlled. _x000D_ _x000D_ Aims_x000D_ 1. Study a key activation process of the APC/C: the interaction between APC/C and the N-terminal domain of its co-activators._x000D_ 2. Identify Nek2A (an early mitotic substrate) receptor in the APC/C to gain a more comprehensive view of substrate recognition._x000D_ 3. Reconstitute recombinant Xenopus holo APC/C for functional and structural studies._x000D_ _x000D_ Method_x000D_ We will use an optimized cell-free system derived from Xenopus eggs that can recapitulate APC/C-dependent substrate recognition, ubiquitylation and proteolysis. We have recently discovered that the co-activator Fizzy/Cdc20’s N-terminus has an independent function in activating APC/C besides its substrate recruitment. Preliminary results already indicate several promising N-terminal peptides with higher affinity to the APC/C and a novel interaction. Combined with site-specific photo-activation, we will identify the receptor subunit and investigate how this interaction is regulated in order to control the APC/C. Similarly, we will identify the Nek2A receptor to deepen our understanding of substrate recognition by the APC/C. Taking advantage of recent success in the Barford lab (ICR, London) in reconstituting budding yeast APC/C, we will reconstitute Xenopus APC/C holoenzyme and sub-complexes and then study the genetically engineered APC/C in egg extract where APC/C or protein kinases have been specifically depleted. This will enable us to discover the role of each subunit and the regulation of the APC/C ubiquitin ligase system. _x000D_ _x000D_ How the results of this research will be used_x000D_ Deregulation of the APC/C has been reported in several cancers (colon and breast cancer). Accumulative evidence also suggests that the APC/C might be a better therapeutic target than the spindle assembly checkpoint that the clinically approved taxane family of anti-tumour drugs activates. Detailed analysis of the APC/C system will lead to a new therapeutic development specifically targeting cancer cells by directly inhibiting the APC/C, thus preventing side effects in normal cells.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Validation of FLT-PET as a predictive biomarker in breast cancer
Funder: Cancer Research UK

Abstract: Background There is a clear need to develop and validate methods for early and objective assessment of tumour response to therapy, to improve patient outcomes and support development of new drugs. Cell proliferation biomarkers could be useful for predicting which patients will respond to therapy, but there are limitations associated with established methods such as Ki67. Positron emission tomography (PET) imaging with [18F]-3’-deoxy-3’-fluorothymidine (FLT) could overcome many of these issues, but FLT-PET needs to be validated to ensure that it is a robust and reproducible method. A few single-centre FLT-PET studies have been performed in the UK, but the high FLT price, limited FLT availability and complexity of running multi-centre studies with experimental PET radiotracers, have prohibited its use in multi-centre trials. There is a pressing need to perform a multi-centre trial with FLT, to validated it as a biomarker of proliferation. This will require identifying and solving the issues associated with setting up and running multi-centre PET trials involving experimental tracers and recruiting enough patients to compare FLT with an established cell proliferation biomarker. Primary Aims Validate FLT as an imaging biomarker of proliferation in primary breast cancer against the known histochemical proliferation marker Ki67 Evaluate whether change in FLT uptake produces important information on FLT as a predictive biomarker of response to AI therapy Identify and solve the issues associated with a multi-centre PET trial with an experimental radiotracer. Methods Patients with tumours bigger than 2cm will be recruited from POETIC, a multi-centre breast cancer trial. A baseline FLT examination will be performed prior to commencing aromatase inhibitor (AI) treatment. Patients will undergo a second FLT examination after 10-14 days of treatment, prior to surgery. Most patients will be imaged using a static PET protocol; SUVmax will be determined 60min after FLT injection. 10 patients will be imaged with dynamic PET. How results will be used Correlation between basal FLT SUVmax and basal Ki67 index will be calculated. A high correlation (80% or higher) will indicate that FLT-PET is a satisfactory surrogate for Ki67. This will allow FLT-PET to be used as a surrogate biomarker of cell proliferation in breast cancer. Dynamic PET data will be modelled and compared with gene expression profiling data to learn more about the factors that modulate FLT uptake. Identifying the issues associated with multi-centre PET trials with experimental radiotracers will make it easier to perform this type of study in the future.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: CRUKD/12/009: FIESTA: FGFR Inhibition in Epithelial Solid Tumours - AZD4547 in combination with gemcitabine and cisplatin: a phase Ib trial in advanced bladder cancer and other solid tumours
Funder: Cancer Research UK

Abstract: Background The combination of gemcitabine and cisplatin (GC) intravenous chemotherapy is a standard of care, both as neoadjuvant chemotherapy for life-threatening, muscle-invasive bladder cancers, and as first-line palliative chemotherapy for metastatic bladder (and other urothelial) cancers. Although bladder cancer is a chemotherapy-sensitive disease, unfortunately there have been no significant improvements in treatment outcomes for advanced bladder transitional cell carcinoma (TCC) for nearly two decades. However, recent advances in our understanding of the molecular pathogenesis of bladder cancer suggest that appropriate targets for novel agents include the Fibroblast Growth Factor Receptor (FGFR) family. Data from our lab, and others, indicate that both FGFR1 and 3 are rational targets for molecularly-targeted therapies, as both play key roles in bladder cancer pathogenesis, and that there are positive benefits from combining the highly-selective FGFR inhibitor, AZD4547 with either gemcitabine or cisplatin. Aims • To investigate the safety, tolerability and feasibility of the novel AZD4547 plus gemcitabine/cisplatin (AGC) combination in advanced non-haematological malignancies • To obtain a preliminary indication of the relative toxicities of AGC and GC. • To obtain information on the pharmacokinetics of AZD4547 in combination with GC • To obtain a preliminary indication of the relative activity of AGC, compared to GC, in locally-advanced metastatic TCC of the urinary bladder (and other urinary tract sites) • To guide the development of a subsequent randomised controlled trial of AGC as neoadjuvant chemotherapy, as a component of radical treatment for muscle-invasive bladder TCC. • To collect blood & tumour samples and to perform functional imaging studies for potential future exploratory research studies, including the identification of predictive biomarkers of response to AZD4547 and/or GC Methods This is a UK multi-centre, open-label Phase Ib trial, consisting of a preliminary, dose-escalation stage, followed by a secondary small, randomised expansion stage. • The dose-escalation stage of the trial (3-6 patients at each of 4 planned dose levels) will be for “all-comers” – patients with advanced solid tumours (including bladder cancer) for whom the GC combination would be appropriate therapy. • The randomised expansion cohort (10 patients in each of 2 arms – AGC vs GC) will be restricted to patients with advanced/metastatic TCC of the bladder. How the results of this research will be used This trial is intended as a preliminary study to guide a potential subsequent randomised controlled trial of AGC vs GC as neoadjuvant chemotherapy (NAC) for life-threatening, but potentially-curable bladder TCC.


Research activity codes: 6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Genetic insight into ethanol and reactive aldehyde induced DNA damage and epithelial cancer development
Funder: Cancer Research UK

Abstract: Alcohol is one of the most common carcinogens that humans are habitually exposed to. On a worldwide scale this chemical is responsible for a large number of cancers. Despite this the mechanisms by which alcohol contributes to cancer risk is largely not known. We have recently found in mice that when we combined the inactivation of a specific step alcohol metabolism with the FA DNA repair defect this precipitated a profound phenotype. Such animals are developmentally compromised, very sensitive to ethanol and unusually prone to cancer. We believe that this points to a fundamental function of the FA DNA repair pathway in protecting our genomes from a byproduct of ethanol metabolism. This chemical is acetaldehyde a chemically highly reactive simple aldehyde. The aim of this grant application is to build on our discovery and establish new mouse models for ethanol-induced carcinogenesis. These models will allow us to address how ethanol and its metabolic byproduct acetaldehyde contribute to certain epithelial cancers. In addition the work will extend to define how other aldehydes, which are also like acetaldehyde omnipresent, and are also byproducts of certain metabolic transactions that might cause DNA damage and cancer predisposition. We have also found that aldehyde accumulation by acute alcohol exposure in FA repair pathway defective mice or cells results in a profound cytotoxic effect. We intend to extend on this observation to determine if aldehyde catabolism can be manipulated both for cancer prevention and also treatment.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: How is mitosis controlled by ubiquitin-mediated proteolysis?
Funder: Cancer Research UK

Abstract: Background: My overall aim is to determine how the mutual antagonism between the Anaphase Promoting Complex/Cyclosome (APC/C) and the Spindle Assembly Checkpoint (SAC) ensure that each daughter cell receives an equal and identical set of chromosomes in mitosis. This is directly relevant to cancer research because when this process fails cells can suffer chromosome damage and become aneuploid, a hallmark of many types of cancer. Furthermore, the therapeutic action of a number of agents, such as the taxanes and the vinca alkaloids, is derived from their ability to activate the SAC. We will build on our research over the past 10 years, which has begun to reveal both how the APC/C and the SAC work, and how we may be able to increase the therapeutic action of these drugs. Aims: 1) To determine how the APC/C is activated at mitosis 2) To determine how the APC/C is inactivated in the next interphase. 3) To determine how the SAC inactivates the APC/C. Methods: We will combine our live cell assay for APC/C-dependent proteolysis with biochemical and mass spectroscopy analysis of SAC complexes and APC/C ubiquitylation activity using 'knock-in' cell lines in which one allele of a specific gene is epitope-tagged. How the research can contribute to understanding cancer: A number of therapeutic drugs act through the control of mitosis through the SAC and its inhibition of the APC/C. Our studies on how the APC/C is activated and inactivated, and how it responds to the SAC could provide insights into better ways to kill cancer cells. Indeed, we have found that we can enhance the ability of taxol and other drugs to arrest cells in mitosis and subsequently promote their death through apoptosis. We hope that this research will provide further insights into how to develop drugs to improve the therapeutic action of these important anti-cancer drugs.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Centres Post-Doctoral Surgical Research Training Scheme
Funder: Cancer Research UK

Abstract: Surgical training associated with the Oxford Cancer Research UK centre: The Oxford Cancer Research Centre builds on the city's long history of groundbreaking scientific research, bringing together world-class scientists, doctors, nurses and radiotherapists to speed progress in beating cancer. The Centre is a partnership between Cancer Research UK, the University of Oxford and the Oxford University Hospitals NHS Trust. Researchers at the Centre are studying many different types of cancer. In particular, they are focusing on breast, skin, gastrointestinal and urological cancers, which covers bladder, prostate, kidney and testicular cancers. Radiotherapy research is a particular strength of the Oxford Centre, spearheaded by scientists at the Gray Institute for Radiation Oncology and Biology - created to be the largest group of doctors and scientists specialising in radiotherapy research in the country. Another key strength is research into the genetic and lifestyle factors that increase the risk of cancer, building on decades of experience in this area. Oxford scientists first showed that smoking causes cancer more than 40 years ago, and researchers in the city have made many more major discoveries in understanding the causes of the disease. Other areas of expertise at the Oxford centre include research into the faulty genes and molecules that lie at the heart of cancer. Researchers will also be investigating ways to improve cancer diagnosis, as well as searching for 'biomarkers' to help doctors predict the effectiveness of cancer treatment. A major aim of the Oxford centre is to bring benefits to cancer patients faster than ever before. To achieve this, the University has established a new Department of Oncology next door to the new Oxford Cancer Centre at the Churchill hospital in the city. This will help to build stronger links between researchers working in the lab and doctors caring for cancer patients, speeding up the creation of new treatments. Finally, the centre is designed to be part of the community in and around Oxford. Working with our Local Engagement and Development team, researchers will engage with local people to raise awareness of cancer and the world-class research on their doorstep.


Research activity codes: 6.4 SURGERY;6.9 RESOURCES AND INFRASTRUCTURE (TREATMENT EVALUATION)
Health categories: CANCER AND NEOPLASMS
Title: Identification and targeting of clinically relevant molecular and genetic subtypes in colorectal cancer
Funder: Cancer Research UK

Abstract: Background: In advanced colorectal cancer (CRC), addition of cetuximab to standard chemotherapy regimen has resulted in modest increases in responses rates and overall survival, but only in patients with Kras wild type (WT) tumours. There are currently no biological agents available that benefit patients with Kras mutant (MT) tumours. In addition, the recent advances in the use of ‘biologics’ in metastatic CRC has not translated into any clinical benefit in the adjuvant setting. The challenge is now to identify efficacious treatments for patient groups with stage II and stage III CRC based on the specific molecular characteristics of the patient’s tumour. Aims: A) Investigate the importance of JAK/STAT3 and EpHA2 as novel modulators of chemo-resistance and metastasis in CRC, particularly KrasMT disease. B) Identify clinically relevant molecular and genetic subtypes of stage II and III CRC using a novel integrated transcriptional profiling and next generation sequencing approach. Methods: Part 1: In vitro cell proliferation, apoptosis and migration studies, in vivo xenograft and orthotopic models, immunohistochemistry, ELISA assays. Part 2: 2A: In the discovery screen, we plan to perform transcriptional profiling on 150 stage II and 150 stage III CRC samples. 2B: Bioinformatics analyses will be carried out to identify clinically relevant molecular sub-groups, and specific sub-groups will be selected for next generation sequencing (NGS). 2C: We will validate prevalence and prognostic role of identified molecular genotypes in an independent large datasets of stage II/III CRC. 2D: We will map the genotypes defined as clinically important to Sanger Institute drug sensitivity database to identify novel treatment strategies for defined molecular subsets of stage II and stage III colorectal cancer. How the results of this research will be used: 1. This work will determine whether targeting JAK/STAT3 and EpHA2 pathways could be novel treatment strategies for CRC tumours, particularly KrasMT disease. In addition, we will determine whether JAK, STAT3 and EpHA2 exhibit potential as predictive and/or prognostic biomarkers in CRC. 2. This work will establish a unique molecular taxonomy of stage II and III CRC by classifying the disease into distinct subgroups based on genetics and gene expression. The prognostic and predictive significance of these subgroups will be evaluated. In addition, by connecting specific molecular and genetic subtypes to drugs, this work will also provide a unique avenue for accelerated drug development for poor prognosis stage II and stage III CRC.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;5.1 PHARMACEUTICALS;4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: CRUK/10/056: Translational research in potentially hormone responsive recurrent/metastatic gynaecological neoplasms - TRANS PARAGON
Funder: Cancer Research UK

Abstract: Background:_x000D_ The PARAGON study is an international phase II study led by ANZGOG investigating the role of the aromatase inhibitor in hormonally sensitive gynaecological cancer. These tumours are relatively rare and this is the first large study to include such patients. Collecting material from these patients will allow the generation of a unique resource for future study._x000D_ _x000D_ Aims:_x000D_ 1 To collect tumour blocks and blood samples from patients entering the PARAGON study._x000D_ 2 To generate tissue micro arrays from these blocks to allow collaborative research investigating the role of hormones in gynaecological cancer_x000D_ _x000D_ Methods:_x000D_ all patients entering the PARAGON study will be approached to give consent to enter TransPARAGON. If consent is given, then blocks from primary surgery and any subsequent surgery for relapse will be collected from the referring centre and transferred to a central biorepository (Glasgow, UK). In parallel blood samples will also be taken for future analysis._x000D_ _x000D_ How the results of this research will be used:_x000D_ 1 to investigate the interaction between androgen receptor expression, estrogen receptor expression and subsequent response to an aromatase inhibitor_x000D_ 2 to generate tissue microarrays to promote collaborative research


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES;4.5 RESOURCES AND INFRASTRUCTURE (DETECTION)
Health categories: CANCER AND NEOPLASMS
Title: Molecular mechanisms by which the cancer chemopreventive properties of transcription factor Nrf2 are regulated through its Neh6 redox-independent phosphodegrons as a consequence of GSK3 activity
Funder: Cancer Research UK

Abstract: Background_x000D_ Transcription factor Nrf2 plays a central role in cancer chemoprevention. It is an unstable CNC-bZIP protein that under normal conditions is rapidly targeted through its N-terminal Neh2 domain for proteasomal degradation by Keap1, a redox-sensitive substrate adaptor for the Cul3-Rbx1 ubiquitin ligase. Many cancer chemopreventive agents are thiol-reactive, and induce cytoprotective genes by inhibiting Keap1 and stabilizing Nrf2. Recently we made the exciting discovery that the Neh6 domain of Nrf2 contains two phosphodegrons that allow SCF(beta-TrCP) to ubiquitylate the transcription factor in a GSK3-dependent manner. However, it is not known which of the potential GSK3 consensus sites are phosphorylated and what other kinases are required to create the beta-TrCP recognition motifs. We now wish to test the hypotheses that the ubiquitylation of Nrf2 by SCF(beta-TrCP) is critically dependent on GSK3 in vivo, and that this in turn requires additional kinases to create the two beta-TrCP binding sites in Neh2._x000D_ _x000D_ Aims_x000D_ 1. Map the GSK3 phosphorylation sites in Nrf2, and identify other kinases that are required to phosphorylate the CNC-bZIP protein in order to create the phosphodegrons._x000D_ 2. Determine how the loss of phosphorylation sites impairs in vitro ubiquitylation of Nrf2 by SCF(beta-TrCP)._x000D_ 3. Establish the in vivo contribution of GSK3 to setting the basal and inducible expression of Nrf2-target genes, and its consequence for cancer chemoprevention._x000D_ _x000D_ Methods_x000D_ Phosphorylation sites in the Neh6 domain of Nrf2 will be identified using mass fingerprinting and SILAC technology. Such sites will be verified using phospho-specific antibodies to probe endogenous Nrf2. The identity of the kinases required to produce the phosphodegrons will be determined by the use of specific inhibitors and by RNAi knockdown. The significance of phosphorylation of the beta-TrCP binding sites will be determined by using immobilized phosphorylated peptides to capture [35S]methionine-labelled beta-TrCP. In vitro assays will be performed to determine residues in Nrf2 that have to be phosphorylated in order for it to be ubiquitylated by SCF(beta-TrCP). Cells and tissues from GSK3-null mice will be examined to determine if loss of the kinase increases basal Nrf2 activity in vivo and also the magnitude of gene induction affected by cancer chemopreventive agents._x000D_ _x000D_ Use of results_x000D_ This project will provide novel information about control of Nrf2 through its Neh6 domain. It will therefore allow additional chemopreventive strategies to be devised that do not antagonize Keap1 but involve inhibition of beta-TrCP activity or the kinases required to create the beta-TrCP recognition motifs.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: CANCER AND NEOPLASMS