NIHRDataInsights/HRCSData huggingface dataset:
Title: The role of haematopoietic stem cell bias in acute leukaemia initiation
Funder: Cancer Research UK

Abstract: Acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) are highly aggressive malignancies that, despite advances in treatment, remain associated with significant mortality and morbidity. Understanding the cellular and molecular etiology of acute leukemias is critical to find new treatment options. Hematopoietic stem cells (HSCs) are the only long-lived progenitor cells in the hematopoietic system and accumulate mutations during ageing. Whereas ALL is most common in pediatric patients, the incidence of AML increases during ageing. Previous studies have demonstrated that murine HSCs (mHSCs) are heterogeneous and during ageing HSCs with lymphoid bias become rare and myeloid-biased HSCs more frequent. The aim of this project is to study the role of HSC lineage bias in determining the cellular lineage of acute leukaemias during ageing. To address this question leukaemogenesis will be induced by MLL (Mixed Lineage Leukaemia) translocations, as these can cause both AML and ALL. First, it will be determined how far the activation of MLL-AF4 in mHSCs with defined lineage bias effect the lineage identity of the resulting leukemias. Parallel, human HSC (hHSC) subsets will be identified by single cell transcriptome analysis, prospective isolated and functionally analysed. Furthermore, the role of MLL-AF9 transformation on hHSC subsets at different age will be investigated. To address the effect of HSC bias on leukaemia phenotype inducible Mll-Af4 knockin mice will be crossed to Hoxb5-CreERT2 mice and Gata1-EGFP mice. Transplantation of single HSCs from these triple transgenic mice will allow the establishment of HSC clones with different lineage bias and subsequently the induction of leukaemia by tamoxifen. Based on results of single cell transcriptome analysis, hHSC subtypes will be identified by cell surface markers, isolated by cell sorting and transplanted into NSGW41 mice to monitor lineage outcome. To test leukemogenesis during ageing, leukaemias will be generated from hHSC subpopulations at different ages with MLL-AF9/EGFP and K-RasG12D/tdTomato lentiviral vector. The outgrowth of EGFP+ and tdTomato+ cells will determine the MLL-AF9 transformation potential at different ages and the frequencies of myeloid and lymphoid outgrowth. Finally, the transduced hHSCs will be transplanted into NSG mice to monitor different leukemia development. Understanding the cellular basis for the age-dependent increase in AML incidence and in particular the effect of HSC ageing is highly important and can lead to the development of improved therapies. Furthermore, the better understanding of the cellular origin of infant leukaemia with MLL can also lead to the improvement of current therapeutic options.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: Study of the role of secreted platelet thiol isomerases in the regulation of platelet function, haemostasis & thrombosis
Funder: Medical Research Council

Abstract: Platelets perform a vital role in haemostasis, although inappropriate platelet activation causes arterial thrombosis leading to heart attacks and strokes. Understanding the mechanisms that regulate platelet function is important for the development of more effective strategies to prevent thrombosis. We have reported that during platelet activation and thrombus formation platelets release ERp5, a member of the thiol isomerase family of enzymes. This protein interacts with the beta3 integrin subunit on the platelet surface, whereupon its enzymatic activity enhances platelet function. Protein disulphide isomerase (PDI) is also known to be secreted from platelets and influence their reactivity. Increasing evidence supports the role of thiol isomerases in haemostasis, and recent data implicate PDI in thrombosis in the mouse. Extracellular PDI has also been implicated in the regulation of tissue factor, which triggers coagulation following exposure and de-encryption at sites of injury. We have recently demonstrated that platelets contain a range of additional thiol isomerases, some of which are also secreted and bind to the cell surface during activation. The aims of this study are to systematically characterise catalytically-competent thiol iomerases that are released from platelets during activation, to establish their roles in the regulation of platelet function in vitro, and to determine the potential contribution of each to the regulation of pathological thrombus formation using intravital microscopy models of thrombosis in the mouse. Using reagents that selectively inhibit thiol isomerase family members, we aim to establish the identities of thiol isomerase substrates on the platelet surface, and begin to understand the means by which they modulate platelet reactivity. Understanding of this paradigm for the regulation of thrombus formation may lead to the identification of new targets for the effective pharmacological suppression of platelet function.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: The role of MYBL2 in the age-related development of blood disorders
Funder: Medical Research Council

Abstract: The haemopoietic system is prone to age-related disorders ranging from deficits in functional blood cells to the development of a variety of neoplastic states. Haemopoietic neoplasms often involve defining cytogenetic abnormalities, frequent amongst which is a deletion in the long arm of chromosome 20 (del20q). MYBL2, a key protein involved in the maintenance of genome integrity, is encoded within the minimum del20q region, suggesting a causative role for the factor in disease onset or progression. I have demonstrated that mice expressing half the normal level of MYBL2 are susceptible to the development of myeloid disorders with age. Moreover, analysis of global expression studies conducted with cells from patients with myelodysplastic syndrome (MDS) confirmed that down regulation of MYBL2 expression levels correlates with poor prognosis (in refractory anaemia with excess blasts (RAEB)), regardless of the cytogenetic abnormality. The importance of MYBL2 in myeloid neoplasms is supported by a strong correlation between MYBL2 RNA levels and expression of a subset of genes related to cell cycle checkpoint control in cells from patients with MDS. This project will utilize the mouse model of MYBL2 haploinsufficiency as a tool to study the molecular mechanisms of initiation and progression of haematological malignancies during ageing. The project will also investigate the molecular mechanisms leading to down-regulation of MYBL2 observed in MDS patients that do not exhibit del20q. If it can be validated that down-regulation of MYBL2 is related to MDS with bad prognosis, then MYBL2 could potentially serve as prognostic biomarker of disease progression and have an impact in prospective therapeutic strategies for MDS patients.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: Role of Early B Cell Factor-1 (Ebf1) in replication initiation of normal B cells and in B-cell precursor acute lymphoblastic leukaemia (BCP-ALL).
Funder: Medical Research Council

Abstract: Early B cell factor 1 (Ebf1) is a DNA-binding protein with a pivotal function in the specification of the B lymphocyte lineage. Mutations in the EBF1 gene or deregulated EBF1 expression are found in a high proportion of BCP-ALL cases, and a direct role of Ebf1 in leukaemogenesis is supported by several mouse models of ALL. By conditional gene inactivation, we have found that Ebf1 is required for proliferation and survival of B cells at virtually every stage of their development. Loss of Ebf1-expression results in a phenotype that highly resembles activation of the replication licensing checkpoint. This cell cycle checkpoint represents a physiologically important mechanism that cells utilize to maintain DNA replication rates under conditions of replicative stress. DNA-damage or depletion of replication licensing factors (e.g. Cdc6 or Cdc7) result in activation of this novel G1 checkpoint that prevents S phase progression until the sufficient number of replication-competent origins is achieved. Cancer cells with a deficient checkpoint progress through S-phase, but are eliminated by a subsequent intra-S or G2 checkpoint. Ebf1-deleted transformed cells die without arrest, similarly to tumour cells depleted of Cdc6 or Cdc7 by siRNA. We have found that transformed, but not primary pro-B cells survive upon exogenous c-myb expression (Gyory, Boller et al. 2012). Thus, the lethal S-phase in transformed cells is overcome by overexpression of c-myb or b-myb, which are known transcriptional inducers of G2/M genes. The aim of this proposal is to elucidate the exact role of Ebf1 in replication initiation, to identify the factor(s) that are responsible for the maintenance of the replication licensing checkpoint in normal B lymphocytes and that are potentially deregulated in human ALL. We are planning to investigate the therapeutic importance of replication licensing inhibition in this disease.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: The Thrombin Pathway in Breast Cancer Dissemination: a potential biomarker and novel therapeutic target
Funder: Department of Health (England)

Abstract: Cancer cells express molecules that trigger clotting. This contributes to the very high rate of deep vein thrombosis (clots in the leg) in cancer patients and is a major cause of death. Conversely, the clotting system helps cancer grow and spread. Patients with evidence of clotting have much worse cancers and release more cancer cells into the bloodstream. Most cancer cells in the circulation are stem cells. Stem cells have the ability to produce daughter cells that are different from the main cancer – allowing the cells to resist conventional treatments. I hypothesis that breast cancer growth and release of circulating cells, particularly stem cells, can be inhibited using anti-clotting drugs. First I shall establish what happens to the clotting system and circulating cancer cells when breast cancer is treated with a conventional cancer drug that we know is working. This will allow me to identify new ‘biomarkers’. Biomarkers detect subtle, early response to treatment without having to wait for a measurable clinical response. This means drugs can be tested in smaller short-term trials. These are cheaper and quicker than conventional clinical trials of cancer drugs. I shall also investigate the effect of stimulating and inhibiting clotting, including using new anti-clotting drug on stem cells grown from breast cancers. I shall be assisted by scientists at the Paterson Institute with international recognition in this technique. Finally,in a double-blind randomised trial of two weeks anti-clotting drug/placebo in early breast cancer patients awaiting surgery, I shall measure changes in known biomarkers and novel clotting biomarkers in the tumour before and after drug/placebo. We will also measure changes in cancer cells and clotting in the blood. This research will not delay patients surgery and is an established trial methodology in our unit.


Research activity codes: 1.4 METHODOLOGIES AND MEASUREMENTS;6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: Randomised controlled trial to evaluate ferric carboxymaltose in the treatment of pre-operative colorectal adenocarcinoma related anaemia.
Funder: Department of Health (England)

Abstract: Background There are 35,000 cases of Colorectal Cancer diagnosed each year in the UK, of which up to 60% are diagnosed with pre-operative anaemia. Untreated preoperative anaemia increases the need for allogenic blood transfusion, and has been shown to increase post-operative morbidity and mortality. Previous strategies to correct anaemia are expensive, poorly tolerated or ineffective. These strategies include allogenic blood transfusion; which places a resource burden on transfusion services, increases peri-operative morbidity and mortality, and has been implicated in the transmission of variant Creutzfeldt-Jakob Disease. Currently patients receive oral iron supplementation on an ad-hoc basis, but this is associated with intolerable side effects and its efficacy remains in question. Aims The primary aim of this study is to determine peri-operative blood transfusion requirements in patients with colorectal cancer related anaemia following randomisation to receive either (i) intravenous ferric carboxymaltose or (ii) oral ferrous sulphate (current best practice). Our secondary aims are to (a) determine the effect on post-operative quality of life in each group, (b) compare peri and post-operative outcomes (morbidity, mortality, length of stay), (c) evaluate the baseline (pre-treatment) and pre-operative markers of iron metabolism (haemoglobin, erythropoeitin (EPO), serum ferritin, iron, and hepcidin). Plan of Investigation A phase IV randomised controlled trial comparing oral ferrous sulphate (control group) and intravenous ferric carboxymaltose. Patient consultation has been critical in supporting our pilot study and lay representation will be provided by the patients members of Beating Bowel cancer UK, to aid the design of trial literature and support the running of the trial. Specifically 116 patients with pre-operative colorectal cancer related anaemia will be randomised to receive either oral or intravenous iron supplementation. Response will be determined by blood transfusion requirement peri-operatively, quality of life, 30 day morbidity/mortality, length of hospital stay. Baseline (pre-treatment) markers of iron metabolism will be reviewed to identify those which predict any response to treatment. Potential Impact A positive outcome from this study would reduce the risk and resource burden of excessive blood transfusion and lead to symptomatic improvement in patients with colorectal cancer. Demonstration of success potentially opens the door to new national treatment guidelines with overall health gain for patients and economic/safety benefits for the NHS as a whole. Negative findings, as has been the case with the pre-operative use of erythropoietin, will enable us to exclude ferric carboxymaltose as a viable single therapy option.


Research activity codes: 6.1 PHARMACEUTICALS
Health categories: CANCER AND NEOPLASMS;BLOOD
Title: The genetic basis and pathophysiology of bone marrow failure syndromes
Funder: Medical Research Council

Abstract: The bone marrow (BM) failure syndromes are a heterogeneous group (including those with the subtypes dyskeratosis congenita, aplastic anaemia and myelodysplasia) of life threatening disorders affecting children and adults. They are unified by the inability of the BM to make an adequate number of mature blood cells and patients die prematurely from infection or bleeding. For one of these syndromes, dyskeratosis congenita (DC), we hold a large unique registry, with samples stored from more than 370 families. In approximately 40% of these DC patients the genetic basis is unknown and in aplastic anaemia and myelodysplasia the primary pathology remains unknown in over 80% of cases. The main aim of the research is to determine the genetic basis and primary pathophysiology of the bone marrow failure syndromes dyskeratosis congenita, aplastic anaemia, and myelodysplasia. To facilitate this goal we will undertake genetic analysis (including those based on next generation sequencing) in families we have collected with two or more affected individuals with aplastic anaemia and/or myelodysplasia/leukaemia, but which do not meet the criteria of known bone marrow failure syndromes. Characterization of the familial forms of aplastic anaemia/myelodysplasia/leukaemia will lead to the identification of new genes and biological pathways that are likely to be important in the idiopathic forms of these disorders. Elucidation of the genetic basis will provide new diagnostic and therapeutic strategies that are highly desirable since treatment remains unsatisfactory for many patients with bone marrow failure.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD;CONGENITAL DISORDERS
Title: Hierarchical organization of haematopoietic stem- and progenitor cell populations during steady state and stress haematopoiesis
Funder: Medical Research Council

Abstract: Recent work from our group and others have provided an improved understanding of the hematopoietic stem- and progenitor compartment, including the identification of novel hematopoietic stem cell (HSC) subtypes and progenitor populations. The challenge we face now is to translate this knowledge into a better understanding of the molecular mechanisms that control the specification of HSC subtypes and the downstream progenitor hierarchy, and to subsequently identify and target the molecular perturbations that cause hematopoietic decline during ageing and subversion of normal differentiation in hematological malignancies. The current program has 3 aims: In the first aim we will use CRISPR/Cas9-based gene perturbation and ectopic expression to identify the transcriptional regulators of key lineage bifurcations, and subsequently combine proteomics and high-throughput perturbation of regulatory elements to identify physical and regulatory transcription factor cross-talk and the resulting gene regulatory networks (GRNs) involved in myelo-erythroid lineage specification. This will include measuring the effect of known leukemogenic mutations on protein-protein interactions and GRN activity, in order to identify putative therapeutic intervention points. In the second aim we will study how HSC fitness and susceptibility to clonal expansion is regulated by ageing. In particular, we will develop improved mouse models of clonal hematopoiesis (CH) that allow reproducible and temporally controlled sub-stoichiometric introduction of relevant mutations (Dnmt3a, Tet2) into HSC. These models will be used to study the role of ageing in clonal hematopoiesis and identify extrinsic cues that promote expansion of mutant HSCs as a strategy to ameliorate CH. In the third aim we will use the sub-stoichiometric introduction of mutations into HSCs to generate an accurate model of myelofibrosis (MF), and in addition use HSC subtype-specific Cre drivers to determine the role of platelet-biased HSCs in disease initiation. In these models the role of the aged micro-environment in MF progression will be studied, and putative extrinsic regulators identified and evaluated for their therapeutic potential in iPSC-based organoid models. IPR from the new CH and models will be protected and commercialised where possible. New putative molecular targets will be explored using MHU spin-offs (e.g. Alethiomics)


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;BLOOD;GENERIC HEALTH RELEVANCE
Title: The genetic aetiology of acute early-onset chemotherapy-induced cardiotoxicity in children
Funder: Children with Cancer UK

Abstract: Anthracyclines are one of the most effective chemotherapy drugs used in many childhood cancers but they can lead to a severe form of heart failure called cardiomyopathy. This risk limits the total dose of anthracyclines that can be used, potentially affecting cure rates. This is particularly relevant in childhood acute myeloid leukaemia (AML) where event-free survival is only 50-60%. Potentially, survival could be improved by increasing anthracycline doses in children shown to be at low risk of cardiomyopathy. Anthracycline-induced cardiotoxicity can be defined as early- or late-onset depending on the time of presentation. “Early-onset” anthracycline-induced cardiotoxicity, occurs in 1-5% of all anthracycline-treated children. “Late-onset” occurs in 10-20% of anthracycline-treated children and represents a major cause of severe morbidity and premature mortality in long-term survivors of childhood cancer. We have identified a unique patient cohort who suffered severe early-onset anthracycline-induced cardiomyopathy and thus have shown exquisite sensitivity to the cardiotoxic effects of anthracyclines. The hypothesis of this pilot study is that such severe early onset anthracycline-induced cardiotoxicity is a monogenic trait, where genetic risk is due to inheritance of a rare but high penetrance genetic variant. Should we identify a gene(s) responsible for anthracycline-induced cardiomyopathy, this will enhance understanding of the underlying pathogenetic mechanisms, aiding both prediction of which children will develop heart problems after anthracyclines and the development of more effective treatment regimens.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;CARDIOVASCULAR
Title: An in vivo model of Burkitt lymphoma allowing therapeutic target analysis and drug testing SNPs in cardiac ABC transporter genes as predictors of anthracycline induced cardiomyopathy in childhood cancer
Funder: Children with Cancer UK

Abstract: Treatment for sporadic Burkitt lymphoma has a high chance of success but at the cost of aggressive multi-agent therapy, resulting in distressing and dangerous side effects and prolonged periods of hospitalisation. Furthermore, for children with refractory or relapsed Burkitt lymphoma, treatment options are limited and frequently unsuccessful. However, whilst there is a need for the development of new treatments, this must be balanced by rigorous pre-clinical evaluation to reduce the risk of unsuccessful early phase clinical trials, which not only expose children to unnecessary risk but delay the development of effective agents. Recent identification of recurrent genomic events in Burkitt lymphoma has provided potential therapeutic targets for further evaluation. It is critical, however, that this evaluation considers the differences between patient-derived lymphomas and cultured cell lines and the role of the host microenvironment. This project aims to develop the tools required to conduct the functional analysis of genomic events in Burkitt lymphoma, essential prior to clinical drug trials. The three principle tools are: 1) a cell line based xenograft model to allow in vivo RNA interference (RNAi) based screening; 2) a real time, patient-derived xenograft model for candidate validation and analysis; 3) development of targeted RNAi screening. Once established, targeted screening of multiple genetic events will analyse the functional contribution of each to lymphoma development in vivo. By identifying those components critical to a lymphoma’s survival/growth, the most effective targets can be selected. Targets can then be validated by RNAi and pharmacological inhibition, using human lymphomas in a representative microenvironment.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CANCER AND NEOPLASMS;CARDIOVASCULAR
Title: Towards Reliable Diffusion MRI of Moving Organs
Funder: Engineering and Physical Sciences Research Council

Abstract: Diffusion Magnetic Resonance Imaging (DMRI) is a clinical imaging technique that has the unique potential to provide clinically-relevant information without the use of ionising radiation or invasive procedures. Although DMRI is often used in brain imaging


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS;CARDIOVASCULAR