NIHRDataInsights/HRCSData huggingface dataset:
Title: To investigate the role of neutrophils in breast cancer progression
Funder: Medical Research Council

Abstract: Pre-clinical evidence suggests that neutrophils are mobilised from the bone marrow in presence of primary breast cancer and act to promote distant metastatic growth (Wculek et al, 2015). Clinical evidence suggests that the neutrophil:lymphocyte ratio has prognostic value, and that circulatory neutrophils in breast cancer patients are different compared to heathy individuals (Wei et al, 2016), (Sagiv et al, 2015). Our collaboration lab has found specific phenotypical changes within neutrophil pools in mice with metastatic breast tumours (unpublished data). We hypothesise that those phenotypic changes are functionally linked to the pro-metastatic activity of neutrophils and that similar changes could be present in human patients. G-CSF is given to patients with early breast cancer to reduce the risk of chemotherapy-related neutropenic sepsis but its effect on influencing neutrophil subsets and their tumourigenic function is unknown. Aim: To investigate the role of neutrophils in breast cancer progression Objectives: 1. To assess how G-CSF affects the function and phenotype of neutrophil subsets in G-CSF knockout mice models of breast cancer 2.To characterize phenotypical changes in circulatory neutrophil subsets in patients with early and advanced stage breast cancer compared to healthy controls 3. To assess how circulatory tumour-educated neutrophil subsets are functionally acting on cancer cells 4. To assess how G-CSF dependent replenishment of circulatory neutrophils can influence neutrophil subsets in patients with breast cancer that have had chemotherapy Methods: Neutrophil subsets will be identified using flow cytometry. Functional characteristics will be assessed with techniques including in-vitro culture with breast cancer cell lines, cell viability assays, immunoassays, proteomics and RNA-Seq. Understanding how neutrophil subsets influence tumour progression has clinical implications for developing new therapies to improve patient outcomes.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: MRI Conspicuity of Prostate Cancer: Investigating Clinically Significant Prostate Cancer Not Detected by Multiparametric MRI
Funder: Medical Research Council

Abstract: This project will utilise an interdisciplinary approach to address challenges associated with multiparametric MRI (mpMRI) conspicuity of clinically significant prostate cancer. Primary objectives are: - Describe characteristics of inconspicuous cancer on mpMRI, link these to mechanisms of inconspicuity, and clarify on the level of clinical significance - Describe characteristics of benign disease that generate suspicious signal on mpMRI and link this to mechanisms of conspicuity - Survey patient perceptions on the role that mpMRI plays in prostate cancer, and their views on the relative risks of false negative and false positive results To address the phenomenon of clinically significant cancer not detected on mpMRI, the PROMIS database will be interrogated. All false negative mpMRI cases will be identified, and each mpMRI will be re-evaluated to assess the impact of: re-reading, varying reporting schemes, assessing levels of quality and artefact, and investigating effects of dominant sequence and lesional position. Further advanced software analysis will be conducted on mpMRI regions of interest containing inconspicuous cancer. Prostate biopsies will be rigorously evaluated in their current state (assessing: Gleason grade, percentage of Gleason pattern 4, presence of stromal gaps, cancer core length) and then will be subject to extensive immunohistochemistry assessment to reveal histopathological composition of inconspicuous cancer. Similarly, cases of false positive mpMRI (from the PROMIS and PRECISION cohorts) will be identified and subjected to comprehensive radiological and histological assessment to determine causation of inappropriately suspicious mpMRI signal. To evaluate current patient perceptions of mpMRI, a carefully constructed survey will be disseminated to men with suspected prostate cancer. Their views on acceptability of false negative and false positive mpMRI will be scrutinised, and focus will be given to which facet they value most.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Defining the roles of fibroblast growth factor receptors on glioblastoma stem-like and non-stem cell populations
Funder: Medical Research Council

Abstract: This project will examine the contributions of fibroblast growth factor 2 (FGF2) signalling to tumour progression in glioblastoma, the most common brain cancer in adults. Glioblastoma has a median survival of only 15 months, and is characterized by diffuse infiltration of the healthy parenchyma, and resistance to therapies, which causes recurrence. The presence of FGF2 in the tumour microenvironment, its correlation with tumour grade in astrocytoma, and functions in self-renewal of brain cancer stem cells, support the notion that FGF2 contributes to malignancy and tumour progression in glioblastoma. But the precise actions of FGF2 remain unclear because the expression and functions of different FGF receptors on glioblastoma cells have not yet been investigated in a comprehensive manner. This fundamental lack of understanding limits the potential of therapies and diagnostics based on FGF2 signalling. Here, we hypothesise that FGF2 acts on different cell populations in glioblastoma (through different FGF receptors expressed on their surface) to promote both cancer stem-like cell self-renewal and tumour growth. We will unravel the cell-type specific expression patterns of FGFRs, and use these to isolate stem-like and non-stem glioblastoma cell populations. Genetic ablation and pharmacological blocking of FGFRs will provide proof-of-concept that targeting specific FGFRs may be therapeutically beneficial. Preliminary data suggests that this may enable development of more tumour-selective therapeutics. Transcriptional profiling will facilitate investigating how expression signatures of these cell populations contribute to intertumoural and intratumoural heterogeneity, and whether these can be used to identify biomarkers and/or methylation-based classifiers to improve patient selection for clinical trials.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: The role of supracellular actomyosin in collective cell migration in vivo
Funder: Medical Research Council

Abstract: Cell migration is essential for development, adult homeostasis and cancer invasion. It has been recognized that cells often migrate in groups and not only as single cells. This collective migration of cells has become an intense area of research in Cell, Developmental and Cancer Biology, as it is essential for both morphogenesis and cancer invasion. However the molecular mechanisms behind this phenomenon are still unclear. It has been assumed that leader cells are the ones that actively drag the trailing cluster and therefore most of the efforts have focused on the front. The alternative explanation that trailing cells drive cluster movement is counterintuitive and would be equivalent to the proverb "putting the cart before the horse". Surprisingly, our exciting preliminary evidence suggests just that. Therefore in this project we will test the challenging idea that collective chemotaxis is driven by the back cells ("cart") making the rest of the cells ("horse") move forward. Specifically, we will test the central hypothesis that a supracellular actomyosin ring contracts at the back of the cluster driving the group forwards. To do this we aim to establish: 1) The significance of actomyosin contractility at the back of the cluster 2) The cellular dynamics by which contractility promotes collective chemotaxis 3) The mechanism by which adjacent cells coordinates their contractility We will use Xenopus neural crest for our in vivo developmental studies. Should our hypothesis prove correct, it has the potential to fundamentally change the current view on collective cell migration. Regardless, however, our findings will have far reaching implications not only in morphogenesis and cancer invasion, but in a multitude of other related biological phenomena.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Deciphering the molecular mechanism of Wnt trafficking in gastric cancer
Funder: Medical Research Council

Abstract: There is an urgent need to understand the cellular signalling network because it is fundamental to the maintenance of a stable state of mature tissue. Deregulation of this network leads to devastating diseases such as the initiation and growth of tumours. The Wnt signalling pathway contributes to this important signalling network and aberrant activation of Wnt signalling is a major driver of gastric cancer growth and metastasis. Despite three decades of Wnt research, we are still not able to explain how Wnt ligands are disseminated in a tissue, which is a prerequisite to control the Wnt network effectively during disease. Recently, our work has revealed the existence of an unexpected transport mechanism for Wnt proteins, which changes the way we understand how ligands spread in tissues: Wnt is rapidly loaded on thin cellular extensions to be transferred to target cells for signal activation. However, the mechanism controlling Wnt signalling protrusions is currently unknown. By closing this gap in our understanding of Wnt transport, we will enable the development of novel approaches to control Wnt signalling in malignancies like gastric cancer. This project will provide new understandings of extracellular Wnt protein trafficking at the molecular, cellular and tissue level. Our hypothesis, supported by preliminary data, is that activating the Wnt pathway in the Wnt producing cells reinforces formation of signal protrusions and thus Wnt signal activation in the neighbouring tissue. This will enable us to understand how Wnt signalling spreads through the tissue to induce proliferation in GC. To test our hypothesis, we will analyse the mechanism of Wnt trafficking and analyse the effect of drugs that limit Wnt signalling protrusions to control tumour growth. The results of this multiscale, multidisciplinary project will provide a step change in understanding how the Wnt signalling pathway operates in gastric cancer.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: PET imaging of ATM using 18F-ATMi
Funder: Medical Research Council

Abstract: Given the increasing number of cancer drugs exploiting DNA damage repair (DDR) defects in tumour tissue, such as ATM inhibitors, there is a clear need for patient stratification, therapy prediction and evaluation tools. Previously, our group has developed several methods for visualising DNA damage and genetic instability (a key hallmark of cancer), based on medical imaging techniques used in routine clinical practice, such as positron emission tomography (PET), targeting DDR proteins such as yH2AX and PARP. ATM is a member of the PIKK family involved in cell cycle regulation and DNA double strand break (DSB) damage recognition and repair. Upon ATM activation it phosphorylates a number of proteins involved in cell cycle checkpoint control, apoptotic responses and DNA repair, including p53, Chk2, BRCA1, and H2AX. To date, no companion biomarkers exist that predict the pharmacokinetic/dynamic behaviour of ATM inhibitors in patients. Such a tool would nonetheless be exceedingly useful to select patients that may or may not respond to the treatment. Therefore, to a known ATM inhibitor (ATMi), we have tagged a radioactive isotope, 18F, while retaining its original chemical structure, to allow visualisation of olaparib biodistribution and ATM expression using a PET scanner. We hypothesize that PET imaging with radiolabelled ATMi can be used to determine the response to chemo- and radiotherapy of cancers. Molecular information about the response to treatment will allow much faster decision making regarding treatment, compared to the anatomical changes such as tumour shrinkage that is used routinely. Also, 18F-ATMi may, in future, be used as an early diagnosis system, detecting the increased genomic instability that accompanies tumorigenesis. Although here we are using PDAC and glioma cells in vitro and in vivo as a model system, given applications of PARP inhibitors in many other tumour types, the approach may be expanded to other disease sites.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Force in Migration: The mechanism of Nuclear Force Coupling driven invasive cell migration
Funder: Medical Research Council

Abstract: Cells migrating through 3D matrices form hybrid adhesion structures, which display hallmarks of both focal adhesions and invadopodia. We have identified an interaction between beta-PIX (ARHGEF7; COOL-1) and Myosin18A (M18A) in 3D matrix adhesion sites. Loss of beta-PIX /M18A abolished cancer cell invasion, but increased matrix degradation and pseudopod extension in collagen matrices. These seemingly contradictory results can be explained by our finding that beta-PIX is part of a betaPIX -M18A-nonmuscle myosin 2A (NM2A) recruitment cascade that is required for adhesion site turnover and nuclear-force coupling (NFC). The nucleus can act as limiting factor in 3D cell migration. Migrating cells need to actively squeeze the nucleus through matrix pores. Knockdown of nuclear envelope actin binding proteins called Nesprin-1 and -2 severely affect 3D cell migration. We have found a reciprocal regulation of Nesprin-2 and beta-PIX /M18A function. We can show using a novel nuclear membrane FRET/FLIM force biosensor, that direct force coupling from 3D adhesion sites to the nuclear membrane through beta-PIX/M18A is required for 3D cell migration. We propose that this mechanism establishes front-rear cell polarity during 3D cell migration through NM2 isoform polarisation. In addition, disruption of nuclear-force coupling inhibited the tension dependant translocation of the pro-invasive transcription factors YAP/TAZ to the nucleus. This proposal will explain how nuclear-force coupling facilitates cell invasion into extracellular matrices in three connected work packages (WP): WP1: Elucidate mechanism and function of M18A mediated NFC and NM2 isoform polarisation in 3D invasive migration. WP2: Identification of molecular signalling pathways required for and affected by mechano-transmission from adhesions to the nucleus in cancer cells. WP3: Effect of nuclear force transduction on cancer cell invasion


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Inactivation of the PRH/HHEX tumour suppressor and its effects on inflammatory signalling in prostate cancer.
Funder: Medical Research Council

Abstract: Transforming Growth Factor beta (TGFb) and other cytokines are secreted by tumour associated macrophages and these cells are important in prostate cancer progression, epithelial-mesenchymal transition (EMT), and metastasis. We have shown that PRH/HHEX inhibits prostate cell proliferation, EMT, and cell migration and invasion in part by activating the transcription of the gene encoding Endoglin, a TGFb co-receptor. Moreover, TGFb down-regulates PRH mRNA and protein levels in prostate cells. This is important because PRH is inactivated by multiple mechanisms in benign prostatic hyperplasia (BPH) and prostate cancer including the accumulation of phosphorylated and transcriptionally inactive pPRH. We hypothesise that PRH activity in prostate cells shapes their response to signals from immune cells and that the inactivation of PRH as a result of signals from immune cells, results in increased cell proliferation and cell migration/invasion and thereby contributes to cancer progression. Our aim is to test this hypothesis and in doing so investigate the importance of immune signals in cancer progression. To achieve this aim we will: 1. Establish a Kenyan archive of blood and urine samples from patients with prostate disorders and an archive of prostate tumour sections to include BPH, prostate carcinomas, and castration resistant carcinomas and document the epidemiologic risk factors and clinical histories associated with these samples. 2. Determine the levels of PRH and pPRH in Kenyan and UK prostate cancer samples using immunocytochemistry (IHC). 3. Determine the inflammatory signals (cytokines and immune cells) that are relevant in the Kenyan population using flow cytometry and IHC. 4. Determine how changes in PRH levels alter the response of prostate cells to signals from immune system cells and how signalling from immune system cells down-regulate PRH activity.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;2.5 RESEARCH DESIGN AND METHODOLOGIES (AETIOLOGY)
Health categories: CANCER AND NEOPLASMS
Title: Targeting natural killer cell receptors for immunotherapeutic benefit
Funder: Medical Research Council

Abstract: The aim of this project is to test and develop natural killer (NK) cell therapy for hepatocellular carcinoma (HCC). We have identified that the NK cell receptor KIR2DS2 recognizes peptide:MHC complexes. This project will exploit this finding and determine whether this feature of KIR2DS2 can be targeted for therapeutic benefit. We have generated preliminary data suggesting that a DNA vaccine strategy targeting KIR2DS2 can be used to activate NK cells and that these have potential to inhibit tumour growth in two different models. The overall goal is to optimise this strategy by: a) Addressing co-stimulation of NK cells using this vaccine, by targeting complementary signalling pathways. We will target TLR activation, IL-15 stimulation and the cell surface receptors, NKG2D. b) Refine the vaccine strategy to bring it closer towards translation to the clinic by: optimising delivery, testing additional vaccine constructs, investigating the phenotype of the NK cells generated, and performing pre-clinical testing. c) Test a novel mechanism for tumour recognition by KIR2DS2+ NK cells, based on a cancer peptide derived from XPO1 that we have identified. For this we will define the HLA-C restriction of this peptide, and then test recognition by KIR2DS2+ NK cells in vitro, and then in an in vivo model of HCC. NK cell therapy is an emerging field. We propose a novel strategy to target NK cells, which may have direct anti-tumour effects or, synergise with immunotherapies that target the adaptive immune system.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS
Title: Tumour virus deregulation of the cell cycle: translational control and function of RGC-32
Funder: Medical Research Council

Abstract: The disruption of cell-cycle control by Epstein-Barr virus (EBV) plays an important role in growth transformation, but the mechanisms involved are not fully defined. RGC-32 is translationally upregulated on B-cell infection by EBV and is deregulated in numerous cancers. RGC-32 may influence cell growth by regulating the mitotic kinases, CDK1 and PLK1, but the molecular details are unknown. We have shown that RGC-32 is essential for the growth of EBV-transformed B cells and that the translational repressor, Pumilio, can control its translation. To decipher the role of RGC-32 in viral and non-viral oncogenesis we will study its regulation, function and structure. We will: 1. Investigate the role of specific RNA-binding proteins and stress granules (SGs) in RGC-32 translation control in EBV-infected cells. We will study this in EBV-infected B cell lines and on primary B-cell infection using immunofluorescence, RNA-FISH, Western blotting and RNAi. 2. Investigate whether EBV latent proteins are responsible for activating RGC-32 translation. We will interfere with the function of EBV latent proteins and pathways activated by them and use knock-out cell-lines to determine the influence of specific EBV proteins on the activity or expression of the proteins identified in (1) that control RGC-32 translation. 3. Determine the role of RGC-32 in the regulation of mitosis and response to DNA damage. Using chemical genetics in tractable epithelial cell lines in which we have shown that RGC-32 promotes mitotic entry, we will examine the influence of RGC-32 on mitotic phenotypes and cell-cycle checkpoints through effects on CDK1 and PLK1. 4. Obtain molecular information on the structure and function of RGC-32 complexes. With recombinant proteins, we will determine the mode, sites of association and effects of RGC-32 binding to CDK1-cyclin B1 and PLK1 using FRET-based binding assays and peptide substrate screening. We will also obtain 3D structures of RGC-32 complexes.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: CANCER AND NEOPLASMS
Title: Phase 1 evaluation of CRISPR-CAR gene edited T cells in relapsed refractory B cell acute lymphoblastic leukaemia
Funder: Medical Research Council

Abstract: Universal T cells aim to circumvent HLA barriers and provide an 'off-the-shelf' alternative to autologous cell therapies. We undertook the first application of genome modified T cells expressing a chimeric antigen receptor (CAR) against CD19 to successfully treat infant B cell acute lymphoblastic leukaemia (B-ALL) in 2015. Cells were transduced with a lentiviral vector to express CAR19 and then further modified using TALENS to disrupt T cell receptor (TCR) and CD52. Genome editing technology is evolving rapidly and we have developed a self-inactivating lentiviral vector that tightly couples CAR expression with CRISPR guides through a hybrid 3' long terminal repeat region (LTR). Subsequent electroporation of Cas9 mRNA results in transient CRISPR mediated DNA cleavage and highly efficient disruption of target loci. Further downstream processing enables depletion of residual TCR+ cells and yields highly homogenous (>99% CAR+TCR-) populations. Furthermore, we have developed an automated process with selection techniques to generate CAR-T cells from umbilical cord blood T cells as well as conventional peripheral blood lymphocyte harvests from healthy donors. In vivo, human-murine anti-leukaemia modelling has confirmed functional integrity of CAR+TCR- cells, and found these were less prone to exhaustion compared to TCR+ cells. Extensive molecular screening for off-target events using Digenome analysis, targeted next-generation and whole-genome sequencing was undertaken. We now propose a Phase 1 study to treat children with relapsed refractory CD19+ leukaemia as a proof of concept application of CRISPR genome modified T cells. The therapy will aim to induce molecular remission & thereby secure eligibility for allogeneic stem cell transplantation. We will assess disease responses, relapse, survival as well as complications including cytokine release syndrome and graft versus host disease. Multiple additional therapeutic applications are envisaged upon completion.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES;6.2 CELLULAR AND GENE THERAPIES
Health categories: CANCER AND NEOPLASMS