NIHRDataInsights/HRCSData huggingface dataset:
Title: Lectin-mediated B-cell receptor signaling in human follicular lymphoma
Funder: Worldwide Cancer Research

Abstract: <p>Many B-cell malignancies are thought to depend on B-cell receptor (BCR) signalling for their proliferation and survival, and mutations causing constitutive activation of BCR signalling have been identified in a subset of non-Hodgkin&rsquo;s lymphoma. However, the mechanisms that drive BCR activation in lymphomas lacking such genetic alterations are not known. Our previous studies revealed a potential new paradigm for antigen-independent BCR activation. We found that <em>N</em>-glycosylation sites introduced into immunoglobulin variable regions during somatic hypermutation carry high-mannose sugars and that these are targets for binding and activation of the BCR by mannose-binding lectins.Remarkably, immunoglobulin variable region <em>N</em>-glycoslation sites are present in essentially all cases of follicular lymphoma (FL) revealing a strong selective pressure for these sequences in this disease. Since this represents a novel stimulating pathway for a wide range of BCRs, it could be a common driver of lymphoma growth/survival.</p> <p><strong></strong>We will investigate the hypothesis that FL-specific mannosylated play an important role in activating proliferation and survival-promoting signalling via the BCR.</p> <p>The project has four specific aims:</p> <p>&nbsp;</p> <p>1. To investigate effects of candidate mannose-binding lectins on survival/proliferation in primary FL cells.</p> <p>2. To investigate effects of candidate mannose-binding lectins on BCR-associated signalling pathways, and the effects of inhibition of these pathways by specific drugs.&nbsp;</p> <p>3. To use a transducible cell line to analyze the influence of lectins on BCR pathways.</p> <p>4. To investigate localization of candidate lectins in FL tissue.</p> <p>&nbsp;</p> <p>These studies will provide important new insight into these common human lymphomas which could be exploited to improve the treatment of this disease. For example, our studies will determine the effect of inhibitors of BCR signalling to determine whether lectin-mediated signalling is also susceptible to these agents and detailed mechanistic signalling studies may reveal factors specifically required for lectin-mediated signalling, providing opportunities for novel therapeutic attack.</p>


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Pten mediated stromal /epithelial interactions in stem cell regulation and tumorigenesis
Funder: Worldwide Cancer Research

Abstract: <p>Deregulation of the PI3kinase pathway is a key molecular event underpinning human colorectal cancer. Previously, it has been assumed that such tumorigenesis is driven out of the epithelial compartment, in part as a consequence of increasing the size of the stem cell/cell of origin pool. However, we have shown that epithelial specific deletion of Pten does not immediately impinge upon either adenoma formation or the stem cell compartment. This raises the possibility that PI3 kinase pathway disruption may regulate the stem cell pool and tumour predisposition in a non-cell autonomous manner out-with the epithelial compartment.&nbsp; Preliminary data presented here show that this is indeed the case, with stromal deletion of Pten leading to changes in the epithelial stem cell compartment, enhanced epithelial-mesenchymal transition (EMT) and ultimately enhanced adenoma formation. In this proposal we wish to explore the mechanisms underlying these processes using genetic tools both <em>in vivo</em> and in organotypic cultures, with a specific aim to investigate the potential role of the Src/Fak pathway in mediating pro-tumorigenic signals from the underlying stroma. Our focus on this pathway derives from preliminary data implicating elevated Collagen-1 signalling from the stroma to its epithelial receptor Integrin B1. In summary, we wish to investigate the mechanisms underlying a new paradigm for PI3 Kinase associated tumorigenesis, which fundamentally alters our understanding of how this pathway may drive disease and how we may approach this pathway both diagnostically and therapeutically.</p>


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Cellular and molecular characterization of cytotoxic tumour-reactive CD4+ T cells
Funder: Worldwide Cancer Research

Abstract: <p>Whereas most studies in cancer immunology have focused in the role and activity of tumour-reactive CD8+ T cells, recent work has underscored the relevance of tumour reactive CD4+ T cells in the orchestration of the immune response to cancer, in some cases enhancing and in others suppressing immunity.&nbsp; Our group recently demonstrated that in addition to classical helper and regulatory function, melanoma-reactive CD4+ T cell can also act as potent direct effectors of anti-tumour immunity through acquisition of cytotoxic activity in vivo, mediating impressive rejection of large fully-established tumours in transplantable and spontaneous mouse melanoma models.&nbsp;Whilst CD4+ Granzyme B+ Cytotoxic T cells (CD4CTLs) have been reported in patients with differing pathologies of inflammatory origin little is yet know about their biology, physiological relevance and potential role during tumour progression and immunotherapy. Consindering the potent anti-tumour activity displayed by these cells we believe it is of great importance to further investigate their potential relevance and application to cancer immunotherapy.</p> <p>&nbsp;Hence, the specific objectives of this proposal are:</p> <ol> <li>To define the network of transcription factors controlling acquisition of cytolytic activity by tumour-reactive CD4CTLs.&nbsp;</li> <li>To explore the impact of manipulation of the transcription factors identified in 1 in the differentiation and function of tumour reactive CD4+ T cells.</li> <li>To identify the cellular signals (i.e. signals through the Tumour Necrosis Receptor family, the B7 co-stimulatory and co-inhibitory family and chemokine receptors) contributing to the activation, differentiation and recruitment of tumour-reactive CD4Trp1 CTLs.</li> <li>To evaluate the presence of cytotoxic tumour-reactive CD4<sup>+</sup> T lymphocytes in human malignancies and characterise the cellular and molecular determinants of cytotoxic activity of these cells.</li> </ol>


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.2 CELLULAR AND GENE THERAPIES
Health categories: CANCER AND NEOPLASMS
Title: Non-small cell lung cancer resistance to erlotinib and regulation of glutathione levels
Funder: Worldwide Cancer Research

Abstract: <p>Lung cancer is the principal cancer killer worldwide with non-small cell lung cancer (NCLC) accounting for approximately 85% of cases. Most patients present with advanced disease and less than 50% benefit transiently from chemotherapy so new therapies are urgently required. Genetic studies are revealing changes in subsets of lung cancer that not only enable stratification but also provide new targets for treatment. For example, about 10% of NSCLC patients have tumours with epidermal growth factor receptor (EGFR) activating mutations that are sensitive to EGFR kinase inhibitors such as erlotinib and gefitinib. These drugs, unlike traditional chemotherapy, induce responses in 90% of patients with EGFR mutant tumours. However, the benefits are usually short-lived due to the development of resistance which in about 50% of cases is associated with a secondary/additional T790M EGFR mutation. Therefore, reversal of this resistance is important to improve survival. Our preliminary NMR-based metabonomic data show that NSCLC cells with the T790M mutation have low cellular glutathione levels compared to erlotinib sensitive cells. Indeed, decreasing glutathione levels in erlotinib-sensitive NSCLC cells rendered them resistant to this compound. Conversely, increasing glutathione levels in resistant cells re-sensitised them to erlotinib. Therefore, elucidating the link between glutathione levels and responsiveness to EGFR inhibitors may reveal novel therapeutic approaches to increase their efficacy and prevent/revert resistance to these compounds. Here, we will investigate changes in the expression or activity of glutathione pathway enzymes that correlate with erlotinib resistance using qPCR/Western blotting and mass-spectrometry-based metabolic flux analysis, respectively. We will determine how these changes are linked to the EGFR T790M substitution by introducing this mutant receptor into erlotinib-sensitive cells. We will then use this knowledge to therapeutically target the glutathione pathway in our&nbsp;<em>in vivo</em>&nbsp;erlotinib-resistant NSCLC xenografts in order to assess the potential usefulness of this approach for translation into the clinic.</p>


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Prostate cancer immunotherapy using PSMA-targeted IL-4-responsive T-cells
Funder: Worldwide Cancer Research

Abstract: Immunotherapy using chimeric antigen receptor (CAR)-engrafted T-cells is emerging as a potent new therapeutic modality for malignant disease. The applicants have developed several CARs and, following pre-clinical work that was supported by AICR, we will initiate our first clinical study in 2013. In that trial, patients with head and neck cancer will receive an intratumoral injection of autologous T-cells that express a CAR directed against the ErbB network. To build on this, a body of pre-clinical work is proposed here that will provide the foundation for a second clinical trial, whereby CAR-engineered T-cells are administered systemically to patients with metastatic castrate-resistant prostate cancer (CRPC). This will be achieved through the iterative optimisation of an immunotherapy (named "PiN-4") in which three transgenes are co-expressed in patient-derived T-cells. All transgenes have been developed and validated by the applicants. The first transgene is a CAR named P28z which couples the binding of PSMA (prostate-specific membrane antigen) to the delivery of a T-cell activating signal, via CD3ζ and CD28. The second transgene is a chimeric cytokine receptor named 4αβ which couples the binding of (weakly mitogenic) IL-4 to deliver a potent T-cell growth signal. We are already using the 4αβ system in our forthcoming clinical trial to expand and selectively enrich CAR-engineered T-cells during ex-vivo manufacture of cell products. Here, we propose to take this system a step further, exploiting the fact that CRPC cells produce IL-4. Thus, we hypothesize that T-cells which co-express P28z and 4 αβ will acquire a selective growth, survival and functional advantage within the tumour microenvironment. To test this hypothesis, T-cells will be imaged in-vivo in real time, through the co-expression of a SPECT imaging reporter gene (human sodium iodide symporter) in the PiN-4 vector. By this means, an optimised vector will be advanced to clinical testing.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Advanced inhibitors of tankyrases-1 and -2 and wnt signalling, critical pathways in cancer
Funder: Worldwide Cancer Research

Abstract: We aim to build on our discovery of inhibitors of tankyrases -1 and -2 (AICR 10-0104) to develop exquisitely selective agents. These will be antiproliferative anticancer drugs through three downstream mechanisms, inhibiting wnt signalling, triggering mitotic spindle malfunction and inducing telomere-shortening crisis in proliferating tumour cells. Tankyrases are PARPs, using NAD+ as a substrate to poly(ADP-ribosyl)ate target proteins. Some tankyrase functions are isoform-specific but many are common to both. Their NAD+-binding domains are very highly homologous. Tankyrases are over-expressed in several clinical cancers. Many human cancers have aberrant wnt signalling; tankyrases regulate this system by acting on axin. Tankyrase-1 is essential for correct structure and function of the mitotic spindle and centrosome, acting on NuMA, and it regulates telomerase activity, acting on TRF-1. The project comprises structure-based drug design of innovative new selective inhibitors, evaluation for inhibition of tankyrases in vitro (with counter-screening against other NAD+-requiring enzymes) and cellular functional studies with the inhibitors. Our potent inhibitors bind at the nicotinamide-binding site. IWR-1 is a compound which binds at an adjacent site unique to tankyrases with limited potency. Using our crystal structures of tankyrase-2 with ligands bound, we have designed new molecules incorporating features to bind at both sites. These designed inhibitors will be synthesised by conventional methods. All target compounds fit Lipinski’s Guidelines. All will be assessed and optimised for appropriate pharmaceutical properties ( e.g . solubility). The primary screen will be an in-house assay for tankyrase-2 activity in 96-well format, with counter-screening against LADH, IMPDH-2 and PARP-1. Potent selective compounds will be assessed in human tumour cells for antiproliferative activity and for action downstream of tankyrases ( wnt , telomere elongation), demonstrating both uptake and functional activity. This project should provide lead compounds for this exciting triple-function target for further development towards the clinic.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Interference between replication and transcription as a mechanism of oncogene-induced DNA damage
Funder: Worldwide Cancer Research

Abstract: Cancer is a disease of genomic instability, characterised by high rates of DNA damage and chromosomal rearrangements that lead to cancer-driving mutations. One source of genomic instability during tumour development is the activation of oncogenes such as Ras, Myc and Cyclin E, which interfere with the proper control of DNA replication, leading to replication-associated DNA damage. Advanced tumour cells maintain high proliferation through loss of tumour suppressors and adaptation to DNA damage, which increases genomic instability but also vulnerability to targeted treatments. This project aims to investigate the role of transcription in oncogene-induced DNA damage. Ras, Myc, and a large variety of other oncogenes either are or activate transcription factors that promote tumour development by altering transcription activities. This underlies the use of transcription inhibitors for cancer treatment, but is also a potential source of genomic instability. The loss of proper transcription control can increase physical interference between on-going transcription and DNA replication machineries, which can lead to DNA damage and genetic recombination. We hypothesise that conflicts between DNA replication and transcription are an important mechanism underlying replication-associated DNA damage caused by oncogenes such as Ras and c-Myc. We further hypothesise that DNA damage response pathways can prevent DNA damage arising from interference between replication and transcription. We will test these hypotheses using oncogene overexpression, small molecule inhibitors and RNA interference in human cell lines. We will investigate transcription-associated replication fork slowing using single-molecule analysis of replication fork progression (DNA fibre analyses). We will characterise DNA damage induced by transcription-associated replication fork slowing using immunofluorescence-based DNA damage assays and pulse-field gel electrophoresis, and we will investigate DNA damage response pathways that counteract the causes and consequences of transcription-replication interference.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Histone variant loading in acute promyelocytic leukaemia pathogenesis
Funder: Worldwide Cancer Research

Abstract: Fusion of transcriptional regulators with heterologous partners as consequence of chromosomal translocations leads to leukaemogenesis. The oncogene of Acute Promyelocytic Leukaemia (APL), PML-RARa bears dominant-negative as well as gain-of-function activities leading to epigenetic changes, deregulation of transcription and ultimately leukaemogenesis. However, the molecular mechanisms underlying PML-RARa epigenetic function are not fully understood. Hugues de The’s laboratory has shown that PML-RARa interaction with the chromatin-associated factor DAXX through its PML moiety contributes to transformation in vitro . DAXX, along with the DNA helicase ATRX, has been recently demonstrated to function as chaperone for the histone H3.3 variant. ATRX appears to regulate DAXX/H3.3 chromatin association. We and others have shown that DAXX promotes H3.3 loading at gene regulatory elements and both DAXX and H3.3 are found at bivalent genes. Our supporting data suggest that PML-RARa regulates the H3.3 chaperone complex not only via DAXX association but also by affecting ATRX expression levels. Notably, H3.3 and its loading machinery are found mutated in human cancer, thus potentially implicating alterations of H3.3 deposition in cancer pathogenesis. Based on these findings, we hypothesise that the H3.3 chaperone DAXX regulates APL leukaemogenesis by facilitating PML-RARa-mediated epigenetic changes. H3.3 loading at PML-RARa-regulated loci would affect histone mark composition and nucleosome stability. To address these hypotheses, we have set the following aims: Aim 1. To study the impact of DAXX loss on APL leukaemogenesis Aim 2. To determine the role of the H3.3 chaperone DAXX in regulation of chromatin remodelling and transcription in PML-RARa-transformed cells This study will provide insights into the role of histone variant loading as a mechanism to promote epigenetic changes in leukaemia.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Generation of novel antibody-based HER2:HER3 heterodimerisation inhibitors
Funder: Worldwide Cancer Research

Abstract: The recognition of HER2 as cell-surface receptor which is over-expressed in ~25% of breast cancers has led to the development of a range of therapeutic agents that interfere with its functioning. Amongst these is trastuzumab (Herceptin®), a humanised monoclonal antibody, which binds to HER2 and prevents the formation of HER2 homodimers. Although trastuzumab has been used to treat over 500,000 HER2-positive metastatic breast cancer patients since its approval in 1998 there is an increasing volume of data showing both primary and acquired resistance to this therapy. Evidence suggests that the upregulation of HER3, another member of the receptor family, following HER2 blockade is a major contributor to this resistance. Thus, there is an urgent need to develop reagents which can directly target HER3 and prevent its dimerisation with HER2, thereby offering a strategy for the treatment of trastuzumab-resistant tumours. In 1995 a novel antibody isotype (IgNAR) was discovered in sharks; unlike mammalian antibodies IgNAR binds its antigen via a pair of highly soluble, single-domain binding regions. We have shown that phage-displayed libraries of these binding regions (VNARs) can be constructed from suitably immunised animals then selected for clones that bind with high affinity and specificity to target. Additionally, via their unusually long CDR3 loop (the main determinant for antigen binding) VNARs can bind to epitopes on the antigen that are inaccessible to conventional antibodies (such as clefts and active sites). We hypothesise that these innate qualities of VNARs can be exploited to generate molecules which can effectively block HER2:HER3 dimerisation and halt the resultant downstream signalling. We will test this hypothesis by raising a suite of VNAR single-domain binding proteins directed against human HER2 and HER3 and subsequently assessing the ability of these domains to inhibit HER2:HER3 dimerisation and limit cancer cell growth in vitro.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: Targeting bone disease in myeloma
Funder: Worldwide Cancer Research

Abstract: Bone destruction is one of the most debilitating clinical features of multiple myeloma and new therapies for targeting this aspect of pathogenesis are urgently required. The main aim of this project is to determine whether Fc receptor-mediated modulation of the myeloid compartment is a viable therapeutic strategy for the treatment of myeloma-associated bone disease. Previous work in our laboratory has shown that Staphylococcal aureus Protein A:IgG complexes (SIC) interact with Fc receptors on monocytes, macrophages and pre-osteoclasts, and are able to effectively target autoimmune-driven inflammation, and the associated bone disease. Moreover, via the down-regulation of RANK and CCR1, SIC directly inhibit osteoclastogenesis in an in vitro myeloma/pre-osteoclast co-culture system. In conjunction with SIC’s ability to inhibit the osteoclastogenic potential of myeloma cells we have found that the resulting cellular environment does not support tumour survival. These findings provide us with new insights that reveal an innovative therapeutic strategy for the inhibition of cancer cell-driven bone disease. However, this has yet to be tested in an in vivo model of multiple myeloma. The proposed studies will use a murine model of multiple myeloma (5TGM1) to determine the efficacy of SIC on myeloma driven osteoclastogenesis, and test our prediction that Fc receptor-mediated modulation of the myeloid compartment will not only inhibit the associated bone disease but also prevent myeloma survival in the usually supportive bone marrow environment. In addition, the mechanisms of action will be interogated to investigate whether or not there are novel pathways associated with this modulation that are valid drug tragets. By achieving these aims we will validate Fc receptor-mediated modulation as a potent strategy for the inhibition of bone disease, therefore providing pre-clinical supporting data to justify this as a new therapeutic approach for the treatment of multiple myeloma.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS