NIHRDataInsights/HRCSData huggingface dataset:
Title: Investigation of the role of Syntaxin 16 and mVps45 in cytokinesis
Funder: Cancer Research UK

Abstract: BACKGROUND:_x000D_ Throughout normal development, and in aberrant conditions like cancer, cells divide by cytokinesis. After replication of the genetic material and formation of two nuclei, the mother cell divides by the formation of a furrow that constricts the cytoplasm, leaving two daughter cells connected by a thin intercellular bridge. Resolution of this bridge (abscission) results in the physical separation of the two daughter cells. Understanding the molecular basis of these processes is an important goal, as it is clear that defective cytokinesis can have profound consequences; many key cytokinesis genes are proto-oncogenes (e.g. ECT2, anillin) and loss of tumour suppressors may promote carcinogenesis by disrupting cytokinesis. _x000D_ Present models suggest that intracellular membrane trafficking is essential for abscission. Secretory and endosomal membrane traffic is re-directed into the intercellular bridge during telophase, and in late telophase vesicles from these pathways accumulate in the intercellular bridge. These vesicles are thought to fuse with each other and with the plasma membrane, thus ‘thinning’ the intercellular bridge and facilitating subsequent abscission. _x000D_ _x000D_ AIMS AND OBJECTIVES:_x000D_ Defining the mechanism(s) by which membrane traffic is re-directed into the intercellular bridge and the mechanism(s) involved in the tethering and fusion of these vesicles are the main objectives of my research laboratory. We have uncovered an important role for the SNARE protein Syntaxin 16, and its interacting partner, mVps45, in cytokinesis. We propose that these proteins constitute an important control node in regulating membrane traffic into the intercellular bridge. We describe a series of experiments aimed at defining how, when and where these proteins act to control the abscission event._x000D_ _x000D_ Specifically we will:_x000D_ i. Determine which trafficking steps are regulated by Sx16 and mVps45 in telophase._x000D_ ii. Determine how the interaction of mVps45 with Sx16 controls abscission._x000D_ iii. Identify which midbody protein(s) are mis-localised upon depletion of Sx16 or mVps45._x000D_ iv. Characterise proteins localised within/on Sx16-positive vesicles in telophase. and_x000D_ v. Ask whether phosphorylation of Sx16 controls abscission._x000D_ _x000D_ METHODS:_x000D_ We will employ siRNA and mutant expression approaches to study membrane trafficking in HeLa cells._x000D_ _x000D_ HOW WILL THE RESULTS BE USED:_x000D_ Our data will provide new mechanistic insight into the process of abscission. A fuller understanding of this important stage of the cell cycle represents an important research goal.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Mechanisms of RAS/RAF-driven oncogenesis
Funder: Cancer Research UK

Abstract: Background_x000D_ Oncogenic mutations in BRAF and RAS are prevalent in human cancers and current evidence suggests that that these mutations are early genetic drivers to which late-stage cancers become addicted. Of the cancer types most relevant to this application – colorectal cancer (CRC) and non-small cell lung cancer (NSCLC) – RAS/BRAF mutations are detected in ~45% and ~22% of samples respectively and thus they are associated with ~7000 deaths per year in the UK in the case of both diseases. Despite the recent advent of some effective biological therapies, treatment of these cancers remains a huge clinical challenge and can only be tackled by undertaking further basic research to identify novel therapies and to design strategies for early detection._x000D_ _x000D_ Aims and Objectives_x000D_ We intend to undertake basic research aimed at identifying novel mechanisms by which BRAF and RAS are involved in driving the cancer phenotype. Specifically we aim to:_x000D_ a) Gain a mechanistic understanding of the role of V600EBRAF in the evolution of serrated CRC._x000D_ b) Investigate the role of secreted proteins and the tumour microenvironment in the induction of senescence by both oncogenes in the lung and gut._x000D_ c) Identify mechanisms by which senescence is overcome to allow further tumour progression._x000D_ d) Utilise Braf and Kras-driven lung tumour mouse models as reagents for identifying biomarkers for early stage lung cancer._x000D_ _x000D_ Methods_x000D_ We will use a multidisciplinary approach to explore the aims outlined above. This will involve the use of cutting-edge technologies including mouse models, proteomics, cell biology, biochemistry and studies of human cancer samples. Although much of our work is based on the utilisation of mouse cancer models, it is believed that this is necessary as a detailed mechanistic understanding of the evolution and progression of cancer cannot be gained in any other way. A fundamental aspect of our work is to translate our findings in the mouse to the human by cross-examining the signaling pathways we identify in primary human tissue samples and cell lines._x000D_ _x000D_ How the results of this research will be used_x000D_ The results of our research will: i) Inform the utilisation of existing drugs for the treatment of RAS/BRAF-driven cancers and identify novel drug combinations, ii) Identify novel targets from which new therapies could be developed for RAS/BRAF-driven cancers. iii) Contribute to the development of early biomarkers for NSCLC, which currently stands as one of the biggest clinical challenges in cancer today.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Investigation of the Molecular Basis of Tumour Radiosensitivity
Funder: Cancer Research UK

Abstract: Delivering a potentially curative dose of radiotherapy is associated with the risk of developing significant side-effects due to the effects of radiation on normal tissues that surround the tumour. Significant gains in the therapeutic ratio of radiotherapy could be made if treatments could be developed that selectively rendered tumour cells more sensitive to IR without altering the sensitivity of normal tissues._x000D_ _x000D_ I plan to identify clinically exploitable targets that may enable tumours to be made more sensitive to radiotherapy without increasing the side-effects associated with treatment. In order to do this I will perform a series of siRNA screens._x000D_ _x000D_ These screens will involve a druggable genome siRNA library using HeLa cells, and a library of 800 kinase genes using NSCLC cell lines containing specific mutations. The aim of this work is to identify novel genes whose depletion induces tumour-specific radiosensitisation. Therefore, for each of the candidate genes identified, I will assess the effects of gene knockdown on a panel of tumour and normal tissue cell lines._x000D_ _x000D_ The clinical significance of those genes confirmed as being involved in tumour cell radioresistance will be assessed by correlating the clinical outcomes of retrospective series of cancer patients with the tumour RNA expression levels of these genes.


Research activity codes: 5.5 RADIOTHERAPY AND OTHER NON-INVASIVE THERAPIES
Health categories: CANCER AND NEOPLASMS
Title: Identification of key drivers of carcinogenesis in endometriosis tissue
Funder: Cancer Research UK

Abstract: Background_x000D_ The mortality for epithelial ovarian cancer has not changed significantly over the past 30 years. It has become increasingly clear that ovarian cancer is a series of distinct diseases with different aetiologies and genetic drivers. Recent work (Hunstman, co-applicant) has identified mutations in ARID1A in 57% of ovarian clear cell adenocarcinoma, (OCCA). Endometriosis is considered a plausible precursor of OCCA and in 2 OCCA cases the identical ARID1A mutation was present in contiguous endometriosis._x000D_ _x000D_ As there are no effective therapies for OCCA there is a pressing need for a model of OCCA to elucidate driver mutation mechanisms and test interventions._x000D_ Key features of predictive models are that the animal is immunocompetent and that the tumour arises from a small population of initiating cells in the appropriate site and tissue of origin. These features maintain the host tissue microenvironment and preserve the interactions between tumour cells and the matrix, vasculature and immune system._x000D_ _x000D_ Aims & Objectives_x000D_ _x000D_ 1) To establish and characterise an autochthonous mouse model for OCCA _x000D_ 2) To test whether other candidate driver mutations in OCCA are necessary for progression of endometriosis to OCCA_x000D_ _x000D_ Methods_x000D_ We have developed a unique model of endometriosis which uses wild-type immunocompetent, gonad-intact mice. The resulting lesions are histologically identical to human endometriosis and critically, treatment with an estradiol antagonist suppresses lesion growth. Donor mice, (K-rasLSL-G12V/Ah-Cre+/+/ROSA26R-LacZ+/+), are steroid-primed to sensitise the endometrium, and a small volume of oil is injected into the uterine lumen provoking a decidual response. The oil contains ß-napthoflavone, which induces cre expression driven by the CYP1A1, (Ah) promotor. This excises the lox-stop-lox flanking the k-rasG12V allele. _x000D_ _x000D_ We already have established “floxed” Arid1a mice and will cross with a reporter line (cre-dependant luciferase) and K-rasLSL-G12V/Ah-Cre+/+ animals. We will monitor the expected transformation from endometriosis to OCCA using serial in vivo bioluminescent imaging. This will be followed by detailed histological analysis and molecular characterisation of lesions as they progress (aCHG and RNASeq). _x000D_ _x000D_ How the results of this research will be used_x000D_ Generation of a clinically relevant animal model of OCCA is vital for the development and testing of therapeutics in this rare but lethal cancer and to inform the design of human clinical trials. We will validate the mouse model by genomic comparison to extensive OCCA datasets and perform in vivo validation of targets from synthetic lethal screens in collaboration with the Vancouver group.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Kaposi's sarcoma herpesvirus: The role of latency transcripts in lymphatic endothelial cell biology and oncogenesis
Funder: Cancer Research UK

Abstract: BACKGROUND:_x000D_ Infection accounts for 20% of the global cancer burden. Kaposi’s sarcoma-associated herpesvirus (KSHV, or HHV8) is an oncogenic virus and the etiological agent of Kaposi’s sarcoma (KS) and of certain lymphoid neoplasms, including multicentric Castleman’s disease (MCD), and primary effusion lymphoma (PEL). Lymphatic endothelial cells and endothelial precursors are the cellular hosts for KSHV, with the majority of cells latently infected and subject to functional changes that lead to KS development. KS is a tractable model for elucidating the role of inflammation in cancer, and for delineating critical pathways involved in virus-driven, and non virus- associated malignancies. KS also provides an entry point to study aspects of normal lymphatic endothelial cell biology and lymphangiogenesis, which is critical during tumour metastases. In particular, our work suggests that KSHV-encoded microRNAs, latent transcripts and virus-induced cellular microRNAs regulate differentiation and key cellular pathways linking inflammation and oncogenesis._x000D_ _x000D_ AIMS & OBJECTIVES:_x000D_ We aim to identify the key molecular mechanisms and transcriptional networks underlying virus-induced cellular reprogramming, maintenance of viral infection and cell transformation. The programme consists of three interrelated sections, which integrate key transcriptional (MAF, beta-catenin) and post-transcriptional (microRNA) regulatory circuits to determine their role in lymphangiogenesis and oncogenesis: 1. The role of MAF in lymphatic endothelial cell fate specification; 2. The functions of the latency-associated region; 3. Functional pathways of viral and cellular microRNAs. _x000D_ _x000D_ METHODS:_x000D_ Using a combination of state-of-the-art molecular and cell biological techniques and approaches, in vitro and in vivo modelling and analysis of clinical material, we will examine the functional impact of the signaling networks nucleated by viral transcripts expressed in lymphatic endothelium, and in KS tumour cells._x000D_ _x000D_ HOW THE RESULTS OF THIS RESEARCH WILL BE USED:_x000D_ Our programme will significantly contribute to understanding the regulatory circuits controlling endothelial cell fate during development and tumour lymphangiogenesis. Our data will extend considerably the current knowledge of the roles that viral and cellular microRNAs play in KS biology and their function in cell fate decisions and key developmental processes, such as cell proliferation and lymphangiogenesis. The pathways we identify will highlight molecular targets that can be used as a basis for novel therapeutic approaches against latent viral infection, and cellular processes critical in non-viral malignancies.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Biomarkers of UVR exposure: tools for determining the relationship between the benefits and hazards of UVR.
Funder: Cancer Research UK

Abstract: Background. Solar ultraviolet radiation (UVR) has both negative effects for health, particularly skin carcinogenesis, and positive effects, primarily vitamin D synthesis. Skin cancer has a high, and rising, incidence in countries with large populations of white skinned individuals, including the UK. Skin type, hair and eye colour are predictive of skin cancer risk, implying protection by constitutive melanin, and differences in DNA repair. UVR is a complete carcinogen and the principal aetiological agent in most cases of skin cancer. However, skin exposure to UVR is also responsible for the majority of the human requirement for vitamin D, with over 50% of the world's population at risk of vitamin D deficiency due to low UVR exposure. Vitamin D deficiency causes growth retardation and rickets in children, and leads to osteopenia, osteoporosis and increased fracture risk in adults. Recently, possible links between vitamin D deficiency and other serious health consequences including increased risk of common cancers, autoimmune and cardiovascular disease have become apparent. These are major global concerns and a source of uncertainty amongst the public and health agencies regarding 'safe' sun exposure and vitamin D status. Cyclobutane thymine dimers (T=T) are formed uniquely by UVR and therefore their measurement represents an ideal means to biomonitor UVR exposure, and DNA damage. Aims 1. validation and qualification of our novel assay for urinary T=T for non-invasive biomonitoring of UVR exposure, following acute UVR dose, time and skin type studies. 2. pilot evaluation of the applicability of UVR biomarkers to study the relationship between benefits/risks of cumulative UVR exposure. Methods. Prospectively study the distribution of T=T, and 8-oxo-7,8-dihydro-2'-deoxyguanosine (biomarker of oxidative stress) in urine samples from humans following UVR dose, time and skin type studies, using UHPLC-ESI-MS/ms Comparisons with tissue levels of damage, the current approach to assessing UVR exposure, and gain in serum 25(OH)D, provide added validation. Perform retrospective analysis of a small pilot study for urinary DNA damage, plus 25(OH)D levels in UVR-exposed populations with different sun-reactive skin types and skin colour. How the results of this research will be used. This study will represent the first assessment of the effect of 'real life' exposure to UVR upon relevant, non-invasive biomarkers of DNA damage, and their association with other biological outcomes of these exposures. This will provide a useful, validated method to biomonitor UVR exposure in humans, plus valuable pilot information to power a future, larger study addressing the UVR risk/benefit question.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: A randomised controlled trial of deferred androgen deprivation therapy ± upfront cryotherapy in men with localised radiation recurrent prostate cancer (RRPC) to evaluate efficacy and tolerability (CROP)
Funder: Cancer Research UK

Abstract: There is significant uncertainty regarding the optimal treatment for patients with radiation recurrent prostate cancer (RRPC). Among the available treatment options, none have been formally tested in RCT. Deferred androgen deprivation therapy (ADT) is one of the commonest management plans. As an interventional option, cryotherapy appears to have the most acceptable efficacy to toxicity profile from uncontrolled case series. In the CROP study, we will compare upfront salvage prostate cryotherapy and deferred androgen ablation. Overall, our study objective is to determine the efficacy of salvage prostate cryotherapy in RRPC and to evaluate its side effect profiles. Specifically, the CROP imaging sub- study aims to apply functional imaging with Magnetic Resonance Imaging methodology to predict tumour response and to detect persistent/recurrent disease following second line prostate cryotherapy. We aim to recruit 216 patients to the CROP Imaging sub-study. Two baseline scans will be performed for 30 patients to determine the reliability/reproducibility of the MRI measurements. The remaining patients will have one baseline scan only. All patients will receive a scan at 6 months post cryotherapy. Diffusion weighted images (DW-MRI) will be obtained using a diffusion-weighted EPI sequence. Multiple b values (ranging from 0 to =1000) will be used. Apparent diffusion coefficient (ADC) values and maps will be calculated from diffusion weighted images. All suspicious areas will be chosen within the prostate to perform quantitative DCE-MRI analysis using a single-compartment Tofts-Kermode model, producing estimates of Ktrans (transfer constant from the plasma to the extravascular-extracellular space, EES), kep (contrast efflux from tumour) and ve (fractional volume of extravascular extracellular space). The change in the enhancement characteristics and ADC values for each tumour following cryotherapy will be assessed. Data obtained from fMRI will be used to to investigate the following research questions: 1. What is the value of fMRI prior to cryotherapy in predicting early treatment failure (at 6 months following treatment)? 2. Do data from fMRI obtained before and after cryotherapy predict the time to treatment failure (or disease relapse) beyond 6 months following treatment?


Research activity codes: 4.4 POPULATION SCREENING;6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Evaluation of Lactate as a Biomarker in Clinical Trials of new mechanism-based anti-cancer drugs using 1H Magnetic Resonance Spectroscopy
Funder: Cancer Research UK

Abstract: Many new targeted anticancer drugs cause tumour stasis rather than immediate regression. Non-invasive biomarkers are therefore important in identifying effective targeting and tumour response in clinical trials. Several novel agents target pathways directly involved in lactate production. In preclinical studies glioma cells treated with PI3k inhibitors demonstrated a 70% lactate reduction, while 50% reductions were seen in melanoma xenografts treated with a novel MEK inhibitor. This project is designed to develop lactate imaging methodology on a clinical scanner, and to test the hypothesis that lactate concentration measured with 1H magnetic resonance spectroscopy is a robust non-invasive biomarker of progression and response in patients undergoing treatment with novel targeted anti-cancer drugs in relevant Phase 1 clinical trials. _x000D_ _x000D_ Direct measurement of tumour lactate is often confounded by intense overlapping lipid signals. Commercial lactate sequences are not available for clinical scanners. We have access to a novel prototype Hadamard-encoded multiple-quantum filtered (MQF) spectroscopic imaging sequence written for the Siemens MRI scanner platform that achieves approximately 1500-fold suppression of lipid relative to lactate. The sensitivity limit for a 5-minute scan is ~ 1 mM for a (2 cm)3 voxel, well below many reported tumour concentrations (e.g. 1.5 – 14 mM in brain, 8-14 µmol/g in head & neck, and 6-13 µmol/g in colorectal tumours). _x000D_ _x000D_ The first year will be spent optimising the MQF lactate sequence to maximise lactate selectivity, comparing experimental MQF lactate signals with theory, writing data analysis software in IDL, and developing a signal quantification method, using signals from tissue water as a concentration reference. The MQF sequence will be evaluated for lipid elimination, lactate sensitivity, localisation efficiency, contamination, motion tolerance, and robustness-of-use in test objects and a pilot group of patients. _x000D_ _x000D_ In the 2nd and 3rd years lactate concentration will be measured in three patient groups_x000D_ a) lymphoma patients before and after receiving standard CHOP therapy, and in which preclinical data suggest that large changes in lactate may be anticipated. _x000D_ b) small pilot groups of patients with metastatic colorectal cancer, ovarian cancer or melanoma where disease stage is likely to correspond to high lactate levels. Where this is the case measurements will be performed before and after treatment with standard therapy in an expansion cohort._x000D_ c) in patients recruited to Phase 1 trials of appropriate targeted new agents whose mechanism of action is known to affect lactate, and where preclinical measurements have shown significant lactate reductions with treatment. _x000D_ _x000D_


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES;4.4 POPULATION SCREENING
Health categories: CANCER AND NEOPLASMS