NIHRDataInsights/HRCSData huggingface dataset:
Title: Development of a rapid in-vitro multi-drug test device to predict response to combined drug chemotherapy in leukaemia patients, before treatment.
Funder: Medical Research Council

Abstract: This project addresses a need for an in-vitro multi-drug test device that predicts response to combined drug chemotherapy BEFORE treatment, to allow selection of best dose and combination for individual Acute Myeloid Leukaemia (AML) patients. Currently, standard dose aggressive chemotherapy is given at diagnosis but up to 40% of patients do not respond. Over 50% of new AML diagnoses are in the over 60's and prognosis is poor. Co-morbidity and intolerance to side effects are seen with standard dose chemotherapy, giving a need for in-vitro tests to predict response to low dose, less aggressive, chemotherapy. We have devised a same-day test to measure response of AML blood or bone marrow samples, to the mainstay AML drug, cytarabine. The test predicts patient response with an efficiency of 83%. Our target product is a in-vitro multi-drug test device that predicts response to COMBINED chemotherapeutics. The device uses bioluminescent bacterial biosensors that give a quantitative measure of the active form of chemotherapeutic drugs within leukaemic cells. Drugs such as fludarabine are used in combination, to increase the effectiveness of cytarabine. Preliminary studies show that our device predicts effectiveness of combined cytarabine/fludarabine chemotherapy. Our technology is accurate, robust and cost effective, with few reagents, suitable for routine laboratory use; end point detection is by low light camera with analysis and interpretation software. The results will inform clinicians quickly on the best treatment options BEFORE patients begin treatment, and reduce subsequent morbidity and mortality. Our development plan is: i) optimising biosensor response and test protocols for combined chemotherapy tests ii) scaling down reagent volume and reducing the number of leukaemic cells per test reaction, to get more tests from a single clinical sample iii) validating on 500 clinical samples with known outcomes to provide data for regulatory submissions.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Mechanism of toxicity of aluminium-based adjuvant (ABA) nanomaterials
Funder: Medical Research Council

Abstract: Aluminium-based adjuvants (ABA) are micron-sized materials which upon dissolution in interstitial fluid form nanometer-sized particles with biological effect. Their mechanism of action in improving the immune response remains to be fully elucidated and these nanomaterials are excellent models for understanding nanotoxicity. The latter is important in that the number and amount of nanomaterials to which humans are exposed during their everyday lives are burgeoning without any sure knowledge of their ultimate biological fate and safety. Understanding of the mechanisms of toxicity of nanomaterials in humans could be determined from a thorough understanding of how ABA enhance the immune response. In preliminary research in which phagocytosing cells were exposed to ABA we identified mitochondria as potential targets of ABA mediated nanotoxicity. Cell fractionation suggested that mitochondria accumulate ABA nanomaterials. Herein we combine a thorough examination of the physico-chemical properties of both current and novel ABA with detailed investigations of their biological effect in phagocytosing cells and in an established rodent model of malignant brain tumours. Physico-chemical characterisations will enable an understanding of the bioinorganic chemistry of these materials in biological milieu which will be used to understand their cellular toxicity and ultimately their application to adjuvant-based immunotherapy. We will apply state-of-the-art fluorescent labelling methods such that we will be able to follow the trafficking of ABA nanomaterials both in cell culture and in an animal model. Once a detailed understanding of the toxic mode of action of ABA is obtained the idea is to optimise similar materials and exposure regimes both for the effective and safe use of ABA in vaccinations programmes but also for targetting cell death in malignant brain tumours which could become a therapeutic off-shoot of an improved understanding of the toxicity of ABA nanomaterials.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Development of tumour-targeted gene delivery systems for intravenous cancer therapy
Funder: Medical Research Council

Abstract: The possibility of using genes as medicines to treat cancers is currently limited by the inability to efficiently deliver therapeutic genes to remote tumours by intravenous administration, without secondary effects to healthy tissues. With the long-term aim of developing an efficacious cancer-targeted gene medicine, we demonstrated that polypropylenimine dendrimers conjugated to transferrin, whose receptors are overexpressed on most cancers, were highly promising, tumour-selective gene delivery systems : their intravenous administration to tumour-bearing mice led to a rapid and sustained tumour regression over one month, with disappearance of 90% of the tested tumours and regression of the remaining ones for the cancer model tested. This treatment was well tolerated by the animals. Building on the promising results already obtained, the proposed project aims to develop new "seek-and-destroy" systems, by modifying the tumour targeting functionalities of the dendrimers as follows: 1) Replacement of the transferrin moiety by other promising tumour-targeting ligands of the same family that have intrinsic antitumoral activity, such as lactoferrin, lactoferricin and Beta-35. 2) Preparation of bifunctional dendrimers: conjugation of transferrin and lysine, conjugation of transferrin and transforming growth factor alpha. The tumour targeting and therapeutic efficacy of these dendrimers will be evaluated on various cancer models, in vitro and in vivo after intravenous administration.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: CANCER AND NEOPLASMS
Title: Defining the role of aberrant O-linked glycosylation in breast cancer
Funder: Medical Research Council

Abstract: Aberrant O-linked glycosylation is seen in many cancers and in 90% of breast carcinomas. Breast cancers and especially those expressing the estrogen receptor overexpress two sialyltransferases, ST6GalNAc-II and ST3Gal-I. This results in these tumours carrying core 1 rather than the core 2 glycans observed in the normal mammary gland. We will: 1.Determine how over-expression of ST3Gal-I contributes to tumorigenesis. Expressing this sialyltransferase in the mammary gland results in earlier development of tumours when mice transgenic for ST3Gal-I are crossed with a model of mammary cancer (PyMT mice). We will determine if expression of ST3Gal-I in the virgin gland alters their development by microscopically comparing morphology to control glands. Mammary glands from ST3Gal-I mice x PyMT mice will then be examined for morphology, glycan expression and immune infiltration using histochemistry, applying the proximity ligation assay when necessary. Changes in signalling will be analysed by co-immunprecipitations and Western blots looking at pathways activated in PyMT tumours, many of which are also activated in human cancers. 2.Investigate how the O-linked glycosylation of two glycoproteins influences tumour growth. Using a breast cancer line engineered to express MUC1 carrying core 2 or core 1 glycans, we will study the effect of these glycans on its interaction with EGFR after EGF stimulation using immunofluorescence microscopy. The transcriptome of the cells lines carrying MUC1 with core 1 and core 2 glycans will also be established. Osteopontin (OPN) acts as an instigator stimulating growth of tumour cells. We will determine the glycosyltransferase expression (by qRT-PCR) and glycomic profile (by mass spectrometry) of cells expressing OPN that can stimulate cell growth and engineer the glycosylation of another cell type by shRNA and/or transfection of glycosylatransferases to mimic this glycosylation. The activity of OPN will be tested in murine models.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Role of BRCA2 in the response to replication inhibitors
Funder: Medical Research Council

Abstract: I will use my expertise in the field of replication fork progression and the DNA damage response to replication inhibitors to further our understanding of the role of BRCA2 and HR in the response to replication inhibitors. We will use Gemcitabine (20,20-difluorodeoxycytidine) as a model replication inhibitor and BRCA2-deficient and BRCA2-complemented normal and cancer cell lines of human and hamster origin. We will use DNA fibre analyses to measure replication fork progression in presence of Gemcitabine and replication fork restart after removal of Gemcitabine. Cells will be labelled with chlorodeoxyuridine (CldU) for 20 minutes followed by incubation with Gemcitabine for periods from 30 min to several hours. Fork stalling and restart will be monitored by adding iododeoxyuridine (IdU) in presence or after washing out of Gemcitabine. Chromosomal DNA will be spread out on microscope slides and labelled DNA detected by immunofluorescence using specific antibodies. We will further investigate the role of BRCA2 in the repair of Gemcitabine-induced DNA damage using immunostaining for DNA damage markers phospho-Serine S4/S8 RPA, phospho-Serine 139 H2AX (gamma-H2AX) and 53BP1 to detect excessive single-stranded DNA and double-strand breaks. We will also measure DNA breaks by comet assay and pulse-field gel electrophoresis. To investigate the role of BRCA2 in cellular sensitivity and resistance to Gemcitabine we will use clonogenic survival and apoptosis assays. To better understand the mechanism of BRCA2 function at stalled forks, we will use a combination of mutant cell lines, siRNA depletion and small molecule inhibitors to investigate the roles of DNA repair proteins (RAD51, BRIP1, PALB2 and MRE11) that functionally interact with BRCA2 during the response to Gemcitabine. The known molecular functions of these proteins will allow us to work towards a model of the molecular transaction promoted by BRCA2 at stalled replication forks.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: The central importance of CXCL8 signaling to inflammatory-based tumour initiation and invasive prostate carcinoma.
Funder: Medical Research Council

Abstract: We will exploit well-established transgenic lines modeling Pten loss in the prostate. Models have been chosen due to their suitability in (i) studying the development of pre-malignant tumours, (ii) modelling inflammatory cells recruitment into Pten-depleted tumours and (iii) studying progression to invasive carcinoma. The importance of CXCL8 to tumour initiation and inflammation-promoted disease progression will be studied through a molecular approach. Cross-breeding of Pten-depleted mice with CXCR2-null mice will model the effects of dampening the microenvironment response to cancer cell-derived CXCL8 signaling. Moreover, using adoptive bone marrow transfer from CXCR2-null mice, we can study in isolation the importance of CXCL8-promoted inflammatory cell infiltrate to disease progression and invasive carcinoma. A pharmacological approach exploiting selective CXCR2 antagonists or a non-selective CXCR1/CXCR2 compound in these models will also be performed. Phenotypic effects will be evaluated by post-mortem histopathology of excised prostate tissue, complemented by gene expression analysis of epithelial and stromal tissue isolated by laser-capture micro-dissection to understand the basis of CXCL8-promoted tumour-stromal communication, and the pathways associated with disease progression in Pten-depleted tumours. In vitro experiments will further characterize CXCL8-promoted tumour-stromal communication using co-culture systems; clones modeling the effects of dose-dependent decreases in Pten expression, conditional re-expression of Pten and reductions of CXCL8 in Pten-depleted cells have already been developed for this purpose. Experiments will study the impact of CXCL8 in amplifying inflammatory responses, upregulating other chemokine pathways to stimulate directional chemotactic cell migration and promoting the protease activity required to underpin the matrix remodelling associated with invasive prostate cancer.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Developmental Clinical Studies - Evaluation of [18F]fluoroethyl triazole labelled [Tyr3]Octreotate analogues for the imaging of neuroendocrine tumours
Funder: Medical Research Council

Abstract: The incidence and prevalence of gastroenteropancreatic neuroendocrine tumours (NETs) has been increasing over the past three decades. Due to the high density of somatostatin receptors (SSTR), mainly SSTR2, on the cell surface of these tumours, [111In]-DTPA-octreotide scintigraphy (SRS) has become an important part of clinical management. More recently, however, [68Ga] labelled octreotate analogues are being increasingly used because of their improved sensitivity and specificity (Gabriel et al., 2007). Fluorine-18 radiolabelled analogues with suitable PK would permit positron emission tomography (PET) with more rapid clinical protocols. Previous work by this group, under a Developmental Pathway Funding Scheme (DPFS) award, designed five structurally-related [18F]-fluoroethyltriazole-[Tyr3]octreotate analogues. Based on high binding affinity and low tumour-to-background noise ratio, within this series and also compared to other related F-18 and Ga-68 SSTR imaging agents, we selected [18F]-FET-betaAG-TOCA as the lead compound to take forward into clinical development. We propose to develop [18F]-FET-betaAG-TOCA clinically via a 2 stage trial design. The initial study will assess the pharmacokinetics (PK), biodistribution and safety of the novel tracer employing 'whole body dynamic PET scanning'; of particular interest will be the optimal time for imaging. Using this information we will construct an appropriate protocol for 'whole body static PET scanning' in the subsequent study. We will then compare the diagnostic efficacy of [18F]-FET-betaAG-TOCA PET/CT to [68Ga]-DOTATATE PET/CT in patients with a histological diagnosis of NET. These clinical studies will be used as the basis for future larger clinical trials and a Department of Health application to establish this tracer as the new clinical standard based on equivalent sensitivity and specificity but improved kinetics and handling, as well as ease of GMP manufacturing than the existing [68Ga]-DOTATATE PET/CT.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Human apurinic/apyrimidinic endonuclease (APE1) is a novel drug target in glioma.
Funder: Medical Research Council

Abstract: Aims: APE1 is important in DNA base excision repair and redox regulation. APE1 is linked to glioma pathogenesis, radio- and chemo-resistance. This proposal aims to investigate the potential of APE1 inhibition in glioma. Objectives: 1) To show that APE1 inhibitors potentiate the cytotoxicity of MMS, temozolomide (TMZ) and ionising radiation under normoxic and hypoxic conditions, including in APE1 knockout cells. 2) To investigate synthetic lethality strategy using APE1 inhibitors in ATM, ATR, PTEN and RAD51 deficient cells and compare to wild type cells. 3) To correlate APE1, ATM, RAD51 and ATR expression in human glioma to clinicopathological and neuroradiological features. Methodology: Tissue culture investigations (MTS and clonogenic assays): APE1 inhibitors will be tested in glioma cell lines [U89MG/SNB19/U373V/TMZ resistant (SNB19VR and U373VR)] for their inherent toxicity. At relatively non-toxic concentrations the ability of APE1 inhibitors to potentiate the cytotoxicity of MMS, TMZ and ionising radiation (0 to 10 Gy) will be investigated. The ability of APE1 inhibitors to selectively target hypoxic cells under hypoxic conditions will be conducted. Similar studies will be conducted in APE1 knockout cells. APE1 and PARP inhibitor (positive control) will be tested for synthetic lethality targeting in ATM, ATR, PTEN and RAD51 deficient cells and compared to wild type cells. Functional studies in cells include: Aldehyde reactive probe, neutral COMET assay, gamma-H2AX immunocytochemistry, FACS analyses, and cell proliferation, migration and invasion assays. Clinicopathological and neuroradiological correlations of APE1 and RAD51 expression in glioma tumours will be studied by immunohistochemistry. Scientific and medical opportunities of the study: This proposal will enhance our knowledge of APE1 biology in cancer and lead to a novel treatment strategy for glioma. Radiobiological analysis to be conducted in this study is a key priority area for the MRC


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: The role of P21/Cdc42/Rac1-activated kinases (PAK) in prostate cancer.
Funder: Medical Research Council

Abstract: To elicit new therapeutic interventions for prostate cancer, we must determine pathways involved in cancer initiation, growth and metastases. Cell signalling pathways are involved in all of these roles. The Rho GTPases Cdc42 and Rac1, are molecular switches to a wide array of cellular functions. The main downstream effector for these are the P21-associated kinases (PAK), which have become of interest in cancer. These pathways are involved in a number of cellular processes including cytoskeletal dynamics and actin polymerization, growth factor and steroid hormone signalling, transcription, mitosis, apoptosis and facilitating the metastatic cascade. There is evidence in a number of different cancer models, particularly breast, that the Rac1/PAK1 pathways help regulate tumourigenesis. In the prostate, Rac1 levels are elevated, particularly in castrate resistant disease, and higher expression correlates with poorer outcome. There is evidence implicating Rac1 as a key mediator for the interaction of prostate cancer cells with bone marrow endothelium during osseous invasion. The evidence for PAK1 expression in prostate cancer is limited but high levels of expression have been demonstrated to relate to Gleason score. In vascular tissue, Rac1 is regulated by the CAMKK2/AMPK axis. This pathway acts as an intracellular metabolic master-switch. In our laboratory, we have established the regulation of CAMKK2 and AMPK through androgen receptor (AR) signalling in prostate cancer cells. This link is beginning to shed some light on the metabolic functions associated with the AR. The aim of this project will be to investigate the role of Rac1/PAK1 signalling in the progression of prostate cancer, including castrate resistant cancer, with reference to the AR and metabolic signalling. It is hoped that the results from these studies will increase our knowledge of the processes involved in prostate cancer progression and metastasis and provide targets for drug intervention.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Interaction between Sox-10 and the tumour suppressor Merlin in tumours of the nervous system.
Funder: Medical Research Council

Abstract: It is estimated that 1/300 individuals will develop a tumour due to Merlin loss in their lifetime. In individuals with neurofibromatosis type 2 or NF2 (incidence 1/35,000) inheritance of one mutant Merlin allele and somatic loss of the second leads to patients developing multiple tumours of the nervous system, typically presenting with bilateral vestibular schwannomas but also meningiomas, ependymomas and gliomas, which require repeated surgery for their removal during the lifetime of the patient. Typical life expectancy for a NF2 patient is less than 40 years. Attempts to model NF2 in mice, using conditional Merlin knockout mice, to develop therapies for the human disease have been difficult because the occurrence and tissue distribution of tumours following Merlin loss are different between mouse and man, and it is proposed there are additional genetic changes in human tumours. Many of the transcription factors that control Schwann cell proliferation and differentiation have now been characterised. These include the transcription factor Sox-10 which controls both axon-Schwann cell interaction and induction of Krox-20, the master regulator of differentiation, and together they control both proliferation and myelination of Schwann cells. Using primary human schwannoma cells we have discovered that Sox-10 is reduced or absent in these tumours in vitro and in vivo. We also find and that restoration of Sox-10 in Merlin null cells restores the ability of Merlin null cells to induce Krox-20 and differentiate. Merlin and Sox-10 are both located on the long arm of chromosome 22 and our analysis shows these two genes are lost together in at least 60% of the somatic mutations occurring in NF2. We propose that loss of Sox-10 function in the human Merlin null tumours is a novel and potential major modifier of Schwann cell phenotype in human tumours and plan to model this both in vitro in human schwannoma cells and in vivo using Merlin and Sox-10 conditional mouse strains.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Dysregulation of tissue-specific bone morphogenetic protein signaling initiates and promotes colorectal carcinogenesis
Funder: Medical Research Council

Abstract: Objectives: Overall - To establish constituents of the BMP pathway's involvement in the initation and progression of human colorectal cancer Specific 1. To identify tissue-specific regulatory regions upstream of Grem1 2. Explore the timing and effect of dysregulation of the BMP pathway in human tissue and colorectal cancer cell lines 3. Assess the contributions of canonical and non-canonical second messenger systems in BMP signal mediation Experimental design 1. Regulatory regions - We will use Formaldehyde assisted identification of regulatory elements (FAIRE) Chromatin immunoprecipitation (CHiP) to identify transcription factor occupancy profiles and chromosome conformation capture (3C, 4C) together with luciferase reporter assays in cell lines to assess the enhancer structure of this key genes upstream region 2. Up and down regulation - Clonal ordering studies, in situ hybridisation and immuno will be used on archival or fresh human tissue to assess the site and timing of BMP constituent dysrgulation. Lentiviral stable transduction of cell lines with doxycycline responsive elements or shRNA will be used to up and down regulate BMP2, 4 and Grem1 to allow assessment of effects on cell morphology, proliferation, clonogenicity. 3. Second messenger systems - We will use immuno and qRT-PCR to assess target gene activity followed by specific receptor and pathway inhibitors to determine the contribution of canonical (SMAD4 dependent) signaling and non-canonical (SMAD4 independent) pathways on BMP signaling mediation. Exploitation: We hope to highlight the role of the mesenchyme in control of epithelial stem cells and establish constituents of the BMP pathway as colorectal cancer initiating and promoting genes in familial and sporadic colorectal cancer. This will help to facilitate the development of novel biomarkers and therapeutic agents


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS