NIHRDataInsights/HRCSData huggingface dataset:
Title: Improving the functional affinity of human anti-glycan monoclonal antibodies (mabs) for cancer therapy
Funder: Medical Research Council

Abstract: Mabs targeting tumours show variable efficacy and improving this is a key goal to this $billion industry. Glycolipids are excellent targets for mabs as they are over-expressed in tumours and are co-accessory molecules for key survival pathways. Blocking these pathways can lead to direct cell killing. Despite the attractiveness of the targets, there are very few mabs recognising glycolipids as they fail to provide T cell help and induce low affinity IgM responses. We have overcome this limitation and produced high functional affinity mouse IgG3 (mIgG3) mabs that show potent in vitro and in vivo anti-tumour responses. These mabs show proinflammatory direct tumour killing without the need for immune effector cells or complement and can reinitiate immune responses in the tumour microenvironment. mIgG3 has the unique ability to non-covalently multimerise on the cell surface through intermolecular cooperativity, resulting in increased functional affinity and enhanced tumour cell killing. This only occurs at high antigen density (tumour) thus reducing off target (normal) toxicity. We have chimerised the mouse mabs to human IgG1 (hIgG1) but they have lost direct cell killing and have reduced functional affinity as intermolecular cooperativity is not present in human IgGs. By crosslinking the chimeric mabs with anti-human Ig anti-serum or by changing one key amino acid residue we have shown that direct killing is directly related to functional affinity. The next stage in development of these novel mabs is to recapitulate the functional affinity of the mouse mabs by introducing intermolecular cooperativity in the chimeric constructs. No one has mapped the mIgG3 cooperativity and the key amino acids that differ between hIgG1 and mIgG3 are not patented giving us a unique position. This project will not only instigate the development of our series of anti-glycolipid mabs but could be used by other groups to improve the therapeutic index of any mab.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: MICA: Early phase clinical trial repurposing ATRA as stromal targeting agent in a novel drug combination for pancreatic cancer (STAR-PAC)
Funder: Medical Research Council

Abstract: Pancreatic cancer (PDAC: pancreatic ductal adenocarcinoma) is a lethal disease. Surgical excision offers the best chance for cure but only is feasible in less than a fifth of patients. Systematic reviews highlight the need for new treatment strategies. No effective neo-adjuvant protocols exist to enable surgical excision for PDAC. We propose a proof-of-concept clinical trial to enable surgical resection for locally advanced PDAC by developing a combination of the state-of-the-art chemotherapy (gemcitabine and nab-Paclitaxel) and repurposing All Trans Retinoic Acid to target the stroma. Desmoplastic stroma and hypo-vascularity impede successful delivery of chemotherapeutic drugs for PDAC. Pancreatic stellate cells (PSC) are critical for the desmoplastic stroma and mediate cancer cell pro-survival and pro-invasive capability through multiple signalling cascades. Quiescent PSC (normal pancreas) store retinol which is lost in activated state (PDAC). This effect is maximal when PSC constitute the predominant component of the cellular mix, as seen in surgical resection specimens. This multi-faceted tumour-stroma cross-talk is unlikely to be blocked effectively by targeting a single pathway. Dampening the tumour-promoting interaction between cancer and stromal cells or, in other words, normalising the desmoplastic stroma, by 'multi-targeting' agents may enhance traditional chemotherapy. Our preclinical data demonstrate that we can both target effectively and measure changes within, the stroma following targeting of PSC, using ATRA. Further data suggest other stromal compartments (immune cells and vasculature) also can be targeted, to the detriment of tumour behaviour, using ATRA. ATRA's effect is potentiated to reduce tumour size in combination with state-of-the-art chemotherapy (gemcitabine and nab-Paclitaxel). Thus, translation of preclinical data into clinical trial exploits the 'co-targeting' of both cancer and stromal compartments in a step-wise development.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS
Title: MSCTRAIL for lung cancer
Funder: Medical Research Council

Abstract: Lung cancer is the leading cause of cancer death worldwide with a 5 year survival of only 10%. The majority of patients present with advanced, incurable disease and existing therapies have significant systemic toxicities with only a modest improvement in life expectancy. Other cancer types frequently spread to the lungs at which stage they too become incurable. Our pre-clinical data shows human bone marrow derived mesenchymal stem cells (MSCs) can be successfully modified to carry an anti-cancer therapy using a lentiviral vector. The anti-cancer therapy is TNF-related apoptosis inducing ligand (TRAIL) which induces death in only cancer cells, leaving healthy cells unaffected. The MSCs engineered to express TRAIL (MSCTRAIL) have been shown to home to the sites of tumours, induce tumour cell death and eliminate or reduce lung metastases in mouse models. We have robust processes for production of MSCs, lentiviral vectors and cell transduction within our fully Good Manufacturing Practice (GMP) licensed facilities. UCLH is a large cancer centre with highly skilled clinical staff. UCL is one of the national Experimental Cancer Medicine Centres with a dedicated Clinical Research Facility for early phase trials and is one of the 9 accredited clinical trials units of the National Cancer Research Institute (NCRI) and one of the UKCRC registered Clinical Trials Units. We propose a 60 month (m) project encompassing final testing of a clinical grade product, followed by a phase I/II trial of MSCTRAIL in 28 patients with metastatic lung cancer predominantly affecting the lungs with one year follow-up. Milestones will be: 1. Complete GMP viral manufacture (13m) 2. Trial approvals obtained and cell manufacture process validated (25m) 3. Complete phase I trial identifying Maximum Tolerable dose (MTD) and safety of dose delivery (36m) 4. Completion of phase II, follow-up, reporting to MRC and MHRA and publication (60m)


Research activity codes: 6.2 CELLULAR AND GENE THERAPIES
Health categories: CANCER AND NEOPLASMS
Title: MICA: Molecular imaging of HER2 receptor expression using PET: mechanistic study of the novel [18F]-HER2-Affibody tracer in metastatic breast cancer
Funder: Medical Research Council

Abstract: 1 in 8 women will develop breast cancer in the UK. In 25% of cases over-expression of the human epidermal growth factor receptor-2 (Her2) is found, which is an adverse prognostic factor and used to guide therapy selection. Tumour biopsies are unreliable and painful. There is an urgent need now for a non-invasive test which can measure her2 expression reliably. Aims:To evaluate the clinical utility of the novel PET probe GE-226 Affibody ([18F]FBA-ZHER2:2891) in patients with breast cancer. Methodology: An initial method development phase will include 8 patients with her2 positive and 8 patients with her2 negative metastatic breast cancer who will dynamic PET imaging over 90 minutes post-injection of GE-226. Radial arterial sampling will be performed to obtain a kinetic input function. Scans will be reconstructed by iterative and filtered back projection algorithms. Mathematical modelling techniques will determine GE-226 binding, delivery, and retention in tumour and non-tumour tissue. Metabolism of GE-226 will be quantified by HPLC. A tissue specimen will be obtained for immunohistochemistry and silver in situ hybridisation testing to quantify her2 expression and amplification. Safety data will be collected and patients will be monitored for any adverse effects as this is a first in patient study. This study will determine the optimum time-point for imaging for the second phase of the study. The second phase will include a baseline static PET imaging study to be performed in 25 her2 positive patients with metastatic breast cancer who are to be treated with trastuzumab- emtamsine (TDM-1) (standard for this group). Tumour uptake will be quantified by SUV, response will be measured using standard CT imaging and RECIST criteria after 3 cycles of treatment. The study will be performed at 4 of the largest breast cancer units in the UK by leading oncologists and imaging physicians. The results could redefine management of patients with her2 positive cancer.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: BAG-1: A novel strategy for targeting the androgen receptor splice variants in castrate resistant prostate cancer.
Funder: Medical Research Council

Abstract: Aims/Objectives: BAG-1, Androgen Receptor (AR), AR spliced variant-7 (ARV7) and Ki67 mRNA and protein expression will be correlated with survival from castrate resistant prostate cancer (CRPC). The BAG-1L:AR/ARV7 interaction and BAG-1 regulation of AR/ARV7 signalling will be determined in prostate cancer (PC) cell lines. The role of BAG-1 in driving castration and treatment resistance in PC will be investigated. The ability of small molecules to reverse BAG-1 function in PC will be determined and assays will be established for drug discovery screening efforts. Methodology: NanoString technology will be used to determine mRNA levels in PC patient biopsies. We have generated novel monoclonal antibodies to ARV7 for use in immunohistochemistry (IHC) and immunofluorescence (IF) . Protein expression will be determined in PC patient biopsies using IHC and IF. Co-immunoprecipitation experiments will characterise the interaction between BAG-1L and AR/ARV7 in PC cell lines (PC3, LNCaP, VCaP and 22RV1). BAG-1L regulation of AR target genes (luciferase reporter assays, reverse transcription polymerase chain reaction and RNA-seq) will be determined. Clonogenic assays will determine the role of BAG-1 in both castration and therapeutic resistance. Scientific opportunities: Establish multispectral imaging and NanoString technology for the use in tissue biopsies from PC patient tumour samples. The fellowship will provide new knowledge into regulation of nuclear hormone receptors providing new experimental tools to explore key physiological and pathological pathways and opportunities for drug discovery efforts targeting ARsv. Medical opportunities: Develop predictive and prognostic biomarkers for the management of CRPC and identify novel therapeutic strategies for treating CRPC. Career opportunities: Pre-clinical drug discovery under international leaders at the world's top rated academic drug discovery unit and highly active and effective drug development unit


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: The role of miR-31 in regulating response of colorectal cancer to EGFR inhibition - relationship to the detriment observed in the New EPOC study
Funder: Medical Research Council

Abstract: Background The New EPOC trial demonstrated detriment with the use of cetuximab in the setting of operable CRLM. Our preliminary data suggest that miR-31 may be predictive of the effect of cetuximab in this study. Objectives To dissect the functional role of miR-31 to enhance our understanding of the effect of EGFR inhibition in CRC and to validate miR-31 as a predictive biomarker. Methodology External validation will be provided by analysing miR-31 expression of tissue from the PICCOLO study of panitumumab in advanced CRC. The molecular targets of miR-31 will be identified using in-silico analyses and experimentally verified using western blotting, qPCR, and 3' UTR reporter assays in vitro, and verified in vivo on TMAs I have constructed. Biological significance of altered miR-31 expression will be explored using standard functional assays with CRC cell lines in which miR-31 has been selectively induced or knocked-down. A mouse-metastasis model will be used to determine the effect of altered miR-31 expression on disease progression in vivo. SCID mice will undergo laparotomy with colonic implantation of CRC cell lines with normal, reduced, or overexpression of miR-31. To assess the effect of EGFR inhibition, mice will be treated with chemotherapy+/- cetuximab. Partial hepatectomy at the time of intra-caecal implantation will assess influence of liver regeneration. The IVIS Lumina III in-vivo imager will be used to non-invasively assess disease progression. This will provide a model for the validation and further dissection of putative targets of miR-31. Scientific and medical opportunities The New EPOC study is not the only trial in CRC to show detriment with the use of EGFR inhibition. A greater understanding of the molecular profiles associated with these adverse outcomes will permit better patient selection for EGFR inhibition, and may offer new therapeutic targets. Lastly it may enhance understanding of the mechanisms underlying CRC progression.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Lynch syndrome-associated endometrial cancer: prevention, screening and prognosis
Funder: Medical Research Council

Abstract: Lynch syndrome (LS) is a tumour predisposition condition caused by mutations in the mismatch repair (MMR) genes. Mutation carriers are at increased risk of several cancers including colorectal (CRC) and endometrial cancer (EC). Establishing a diagnosis of LS has the potential to save lives. Intensive cancer surveillance by colonoscopy and removal of colorectal polyps prevents CRC-related death. Prophylactic hysterectomy prevents EC. Benefits extend to other family members who undergo genetic testing/cancer surveillance. The prevalence of LS in EC patients is unknown. We do not know whether screening for LS is feasible, acceptable to women or cost effective. We do not know if the prognosis of LS-associated EC differs from sporadic EC. There is also no evidence to support progestin chemoprevention or EC screening in LS, even though both strategies are routinely recommended. Objectives: 1. To determine the prevalence of LS in EC and the feasibility, acceptability and cost effectiveness of screening for LS in EC 2. To compare prognosis and tumour biology of LS-associated and sporadic EC 3. To assess EC rates in LS patients undergoing screening for EC, progestin chemoprophylaxis and prophylactic hysterectomy Methodology: 1. I will assess 400 EC for microsatellite instability (MSI) and MMR protein loss. Patients with tumour studies suggesting LS will have reflex genetic testing. I will establish the prevalence, feasibility and cost effectiveness of LS screening in EC patients. 2. I will compare MSI, MMR protein loss and other EC biomarkers in 165 archived LS-associated EC and stage-matched contemporaneous sporadic EC controls. I will compare EC-specific survival in the two groups. 3. I will conduct a questionnaire survey to assess the prevalence of EC risk-reducing strategies in women with LS (n=364). I will compare EC rates in women who had a prophylactic hysterectomy with those who did not, age-matched and stratified by risk-reducing strategy.


Research activity codes: 4.4 POPULATION SCREENING
Health categories: CANCER AND NEOPLASMS
Title: The effect of AIP mutations on the apoptotic RET pathway in pituitary adenomas
Funder: Medical Research Council

Abstract: AIP-related pituitary disease is a novel clinical entity; however, the mechanism by which AIP, a ubiquitously expressed putative tumour suppressor gene, causes adenomas primarily from a single cell type is currently unclear. RET is a dual function protein with both tyrosine-kinase receptor and, in the absence of ligand, apoptosis-inducing 'dependence receptor' activity. RET is predominantly expressed in growth hormone cells of the pituitary gland and previous data show an interaction between RET and AIP. The current project will study the impact of AIP dysfunction on RET pathway components. Our preliminary data show that knockdown of AIP or overexpression of mutant AIP can inhibit RET-induced apoptosis. Building on this work, we propose to set out and dissect the impact of AIP dysfunction (knockdown and mutated AIP overexpression) on members of the RET pathway (RET, IC-RET, Caspase 3, PKCdelta, CREB, JNK, Pit-1) ) in rat pituitary cell line GH4C1, and dissect the exact point in the pathway affected by AIP dysfunction. We will also explore if AIP, as a partner to heat-shock protein 90, plays a 'chaperone' role in RET biogenesis, trafficking or degradation. In addition to cell line work, we will use primary rat and mouse pituitary cultures expressing endogenous RET to confirm the observed changes in a more physiological setting. Human pituitary adenoma samples from patients carrying AIP mutations will be studied for apoptosis and RET pathway member expression compared to sporadic pituitary adenoma samples. We have developed a pituitary-specific transgenic AIP knockout mouse model and we will study embryonic and postnatal pituitaries for RET pathway member expression and phosphorylation status. Finally viral constructs containing mutated AIP will be microinjected into neonatal and pubertal rat pituitary glands to see the effect on growth hormone cells, hyperplasia and tumour formation and body growth during puberty and adulthood.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Cysteine oxidation as a regulator of cancer cell motility and invasion
Funder: Medical Research Council

Abstract: Cell motility is regulated by a number of signalling pathways that influence the activity and localization of proteins, often via post-translational modifications (PTM) such as phosphorylation. Although oxidation of cysteine residues has been shown to be an important regulator of some proteins, for example protein tyrosine phosphatases, there is little information regarding the identities of proteins modified by cysteine oxidation that contribute to cell migration. We found that during cell migration, hydrogen peroxide (H2O2) is elevated close to the plasma membrane and throughout the cell cytoplasm by fluorescent lifetime imaging microscopy (FLIM). Migration also increases protein oxidation as determined by the incorporation of the sulfenic acid reactive compound dimedone. Using mass spectrometry (MS) we identified over 200 dimedone labelled proteins and determined that oxidation of Cofilin1 in particular is increased in migrating cells. By biochemical and cell biological analysis, we determined that the oxidation of Cofilin1 inhibits its filamentous actin severing activity and facilitates directional cell migration. Using mass spectrometry, we will identify proteins undergoing cysteine oxidation during cell migration across a number of tumour cell lines, and in specific instances will determine how these modifications affect protein function. By siRNA mediated knockdown, we will examine how NADPH oxidase complexes contribute to H2O2 production during cell migration and determine the role of the Rac1 GTPase. Finally, using intravital microscopy to do FLIM measurements of fluorescent biosensor proteins, we will determine how H2O2 production and Rac1 activation are associated with tumour cell invasion in living tissue. These assays will also be used to ascertain whether blocking H2O2 generation is part of the mechanism of action of Rac1 inhibitors in reducing tumour cell invasion.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS
Title: Innovative Approaches for characterisation and treatment of Olfactory Neuroblastoma
Funder: Medical Research Council

Abstract: Olfactory neuroblastoma (ONB), a rare sinonasal tumour, is advanced at diagnosis in a third of patients, in whom prognosis is poor (disease free survival 1.5y). Paediatric disease is frequently aggressive, and the current gold standard treatment (surgery plus chemoradiotherapy) results in significant long-term sequelae. ONB has a strikingly high recurrence rate of 60%, meaning that alternative treatment options and prognostic biomarkers are urgently required. The sparse data available on ONB suggests that the mTOR/MAPK and SHh signalling pathways play a role in tumourigenesis. In cancers, cross-talk between pathways provides a mechanism of drug resistance and treatment failure when single agent inhibitors are used against many different tumours. Targeting both pathways therefore makes rational sense. Aims: i) Establish an inhibitor combination that causes simultaneous pathway inhibition in ONB; ii) RNA seq to identify novel targets and biomarkers; iii) Validate findings on fresh tissue and iv) Reproduce effective treatment in a nude mouse model. Objectives: Using human ONB cell lines, we have established that inhibitors to both pathways reduce cell viability in a dose and time dependant manner. Comparison of multiple treatment combinations suggests novel S6K1 inhibitors plus growth factor receptor inhibition with SHh inhibitors is highly promising. Further outcome measures of efficacy will include western blot, RT-qPCR, apoptosis and cell cycle analysis. RNA seq (n=4 samples, 4 controls) will examine gene expression data, and significant candidates will be validated in overexpression/siRNA cell line studies and on both fresh and fixed tissue samples. In vivo assessment of the optimised drug combination in nude mice will validate the efficacy and assess tolerability. Establishment of a safe, effective, targeted, combined therapy could then be proposed for clinical trials, ultimately aiming to treat both primary and recurrent / inoperable disease.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS
Title: Defining the impact of secondary mutations in CEBPA-mutated acute myeloid leukaemia.
Funder: Medical Research Council

Abstract: The development of leukaemia occurs as a result of the step-wise acquisition of mutations within a haematopoietic stem or progenitor cell (HSPC). It is now recognised that between 5 and 20 mutations occur per AML before the development of frank leukaemia. The CEBPA gene is mutated in 10% of AML and defines a distinct subset of patients. Intriguingly, in a few reported kindreds, mutation in CEBPA occur in the germline, providing clear evidence that CEBPA is the initiating event. The additional mutations that lead to the development of AML remain unclear. Mutations in the methylcytosine dioxygenase TET2 have recently been reported to occur in concert with CEBPA mutation. We hypothesize that combined mutation of CEBPA and TET2 results in epistatic effects on HSPC, which drive clonal evolution to AML. In Aim 1 of this proposal we will utilize zebrafish models to define the effects of combined mutation of cebpa and tet2 on haematopoiesis and leukaemogenesis in vivo. This model is ideally suited to high throughput assessment of the effects of genetic combinations given its size, fecundity and optical clarity. Other important secondary driver events in CEBPA mutated AML are still to be defined. In Aim 2. of this proposal we will capitalise on next generation sequencing data from cases of familial CEBPA mutated AML along with available published whole exome sequencing in CEBPA mutated AML in order to define a series of candidate secondary driver mutations. These candidates will be then be sequenced in a large cohort of 120 patient samples with CEBPA mutations. Mutational profiles will be correlated with clinical outcome data to define clinically relevant recurrent genetic combinations. Mutations identified in this aim will then be poised for functional assessment in our zebrafish model.


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