NIHRDataInsights/HRCSData huggingface dataset:
Title: Specific targeting of a pro-inflammatory B cell subset in rheumatoid arthritis.
Funder: Medical Research Council

Abstract: The importance of B cells in the pathophysiology of rheumatoid arthritis is highlighted by the clinical effectiveness of B cell-depleting therapy (BCDT) using rituximab, an anti CD20 antibody(1). Potential mechanisms by which B cells may drive disease processes in RA include their ability to produce autoantibody, act as antigen-presenting cells, or secrete cytokines. The best characterized autoantigens in RA are posttranslationally citrullinated proteins. The major disadvantage of currently available BCDT is that it indiscriminately removes the vast majority of B cells without specifically targeting those that are involved in pathology. In a systematic study of synovial cytokine expression we have identified a pro-inflammatory B cell population that expresses high levels of RANKL and TNF at the site of inflammation, in the synovial fluid and tissue of patients with RA (2, 3). In an extensive study of the immunophenotype of these cells we have established that they are characterized by surface expression of FcRL4, a transmembrane protein that is related to Fc receptors (3). FcRL4+ B cells express high levels of the costimulatory molecules CD80 and CD86 (3, 4). Expression of FcRL4 is highly restricted to this B cell subset and FcRL4 mRNA expression was detected in inflamed but not in uninflamed synovium. Due to their expression of pro-inflammatory cytokines, their capacity for costimulation and their recognition of citrullinated autoantigens we propose that FcRL4+ B cells are a promising clinical target. We plan to use a custom service offered by AbD Serotec under license from Morphosys to generate human antibodies and use them to target FcRL4+ inflammatory B cells in vitro and in mouse/human chimeric models. Starting with a range of antibodies, we will select the most promising clones over a series of in vitro and in vivo experiments. These experiments will provide data that will allow us to assess the potential of FcRL4 as a target for a novel therapy for RA.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: BIOMARKERS AND MECHANISMS OF FOOD ALLERGY AND ORAL TOLERANCE IN IgE-SENSITISED CHILDREN
Funder: Medical Research Council

Abstract: The majority of children with food-specific IgE do not have food allergy (FA). In equivocal cases, oral food challenges (OFC) are required. However, these are expensive, time-consuming and not without risk. Aims: 1. To improve the diagnosis of food allergy using the basophil activation test (BAT) 2. To validate the mast cell activation test (MAT) and the inhibition of MAT (IMAT) as novel biomarkers of food allergy and food tolerance 3. To understand the mechanisms underpinning FA and tolerance in IgE-sensitised children Methodology: Following the success of BAT in accurately diagnosing peanut allergy (PA) and reducing the need for oral food challenges (OFC), I will create a new BAT to diagnose cow's milk, egg, sesame and cashew nut allergies. I will validate MAT and IMAT as biomarkers of PA and peanut tolerance, respectively. In order to understand the discrepancy between clinical allergy (and tolerance) and the levels of IgE (and IgG4) to peanut allergens, I will test the following hypotheses: 1.The location, number and combination of IgE epitopes in the allergen molecules play a role in the ability of allergen-IgE complexes to trigger effector cell activation; 2.Relative titres and matching of epitope-specific antibodies between IgE and IgG4 contribute to the overall effector cell activation or inhibition. IgE and IgG4 binding to peanut allergen peptides will be performed. IgE and IgG4 epitopes will be identified on the allergen structure and compared between allergic and tolerant patients. The crystal structure of IgE-allergen complexes will allow identifying critical epitopes for IgE binding and their relevance will be assessed on MAT using wild-type and allergens mutated to remove specific epitopes. The blocking effect of IgG4 and Fab fragments with the same specificity as IgE will be tested in the IMAT. Scientific and medical opportunities: This study will improve FA diagnosis and may identify targets for definitive treatment and prevention of FA.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: The roles of novel mammalian insulator factors in T-cell development
Funder: Medical Research Council

Abstract: Insulators are thought to play important roles in establishing cell type-specific transcription patterns and in genome organisation. Mammals are widely believed to rely on a single type of insulator, defined by the factor CTCF. However, recent studies, including my own work, indicate the distribution and activity of CTCF are not sufficient to explain the compartmentalisation and three-dimensional organisation of the genome, implying significant contribution from alternative insulators. Identifying these elements and the factors that mediate their functions is essential for understanding how gene expression programs are established, maintained and changed during mammalian development. Using developing T-cells as a model system, I intend to carry out a combination of genome-wide correlative analyses and functional studies to investigate the roles of newly identified putative insulator factors in transcriptional regulation and chromatin organisation. My research plan presents an opportunity to expand the repertoire of mammalian insulators beyond the current monopoly of CTCF, and gain new mechanistic insights into the roles of these regulatory elements in mammalian development. Drawing up the complete landscape of mammalian insulators has direct implications on our ability to understand and diagnose human disease, as recent comprehensive analyses of genome-wide association data revealed more than 50% of disease- and trait-associated genetic variants lie in intergenic regions bearing hallmarks of long-range gene regulatory elements such as enhancers and insulators.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Investigating NF-kappaB p50 phosphorylation and the regulation of transcription
Funder: Medical Research Council

Abstract: NF-kB is the master regulator of the immune response and is essential for the development and homeostasis of the immune system. The NF-kB subunit p50, encoded by NFKB1, is the transcription factor most highly expressed in human macrophage and is a critical factor in human inflammatory disease. Functional polymorphisms in NFKB1 are significant risk factors for inflammatory disease and cancer. Whilst the importance of NF-kB in human health and disease is indisputable, we lack fundamental information on the biochemistry of NF-kB activity. Phosphorylation of NF-kB is critically important for the transcription of target genes. The impact of specific NF-kB phosphorylation events is highly selective and directs transcription in a gene-specific fashion. However, a comprehensive analysis of p50 phosphorylation and its role in regulating transcription is notably lacking. We propose to generate a unique dataset of transcriptional events that are dependent on site-specific p50 phosphorylation, using a human macrophage model. Our approach will develop a series of gene-expression signatures predictive of functional classes of p50 phosphorylation, the activity of the kinase responsible, and determine how these influence overall transcriptional responses in macrophages. Existing transcriptomic data obtained from human macrophage or tissue from patients with inflammatory disease will be interrogated with phospho-p50 specific transcriptomic signatures. We will generate a co-expression network model of p50 target genes regulated by site-specific phosphorylation. The network will integrate public data on chromatin modifications from human monocytes, macrophages and dendritic cells. The model will provide a powerful tool for the development of novel therapeutic strategies based on the regulation of gene expression by p50 phosphorylation. This proposal will enable a highly innovative approach to tackle fundamental questions in NF-kB biology of high importance to human health and disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: MAximizing Sle ThERapeutic PotentiaL by Application of Novel and Stratified approaches (MASTERPLANS)
Funder: Medical Research Council

Abstract: Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease affecting approx. 16,000 people in the UK. It is highly heterogeneous: each patient presents with a unique fingerprint of organs involved, severity, and response to therapy. The clinical heterogeneity is paralleled by a range of underlying genotypic and immune function abnormalities. A number of new therapies, with different mechanisms of action, have been developed. Overall response rates are only 40-60% but subsets of patients respond very well to each drug. Current therapy is based on a 'trial and error' approach often involving glucocorticoid co-therapy. Delayed and poor control of inflammation results in organ damage, cardiovascular disease and glucocortocoid toxicity. We aim to fundamentally improve care for SLE patients by identifying key endotypes associated with remission and low disease activity (LDA) on particular therapies. Our goal requires a co-ordinated and co-operative approach. Our consortium combines expertise from academia with pharmaceutical and small and medium enterprise (SMEs) partners to identify predictors of response to exemplar therapies. Clinical and genotypic predictors of response will be sought by analysis of existing data from large studies. Two parallel prospective studies assessing tissue, blood and urine from patients with skin or renal lupus will study immunophenotypic predictors of response. These discoveries will be used to develop near-patient devices and algorithms to select therapy in clinic. We will test the benefit and cost-effectiveness of this stratification approach in trials. Using the right drug at the right time will improve control of inflammation for people with SLE. Long-term benefits will be improved survival, improved quality of life and reduced glucocorticoid requirements, whilst providing financial savings for health and social care. MASTERPLANS will be the international model of best practice in delivering stratified medicine to SLE patients.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;6.1 PHARMACEUTICALS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: The role of P2X7 in Autoimmune Vasculitis
Funder: Medical Research Council

Abstract: Antinuclear cytoplasm antibody (ANCA) associated vasculitis (AAV) is an important cause of morbidity and mortality causing multisystem organ damage, including glomerulonephritis (GN) with subsequent progression to renal failure, and life-threatening lung haemorrhage. Better-targeted are needed. The P2X7 receptor (P2X7R) is an ATP-sensitive cell surface receptor with multiple roles, including inflammatory cytokine production and release, apoptosis and antigen presentation. P2X7R mediates renal injury in immune complex mediated GN. However, antibody deposition is not a prominent feature of pauci-immune GN in AAV, the role of P2X7R as a novel therapeutic target in AAV remains to be investigated. We hypothesise that P2X7R is critical in the production of ANCA and leucocyte activation leading to glomerular and lung injury. We aim to investigate a role for P2X7R in patients with AAV and also in a model of myeloperoxidase (MPO) induce experimental autoimmune vasculitis (EAV) in Wistar-Kyoto (WKY) rats. The objectives of the project are to investigate: 1. P2X7R expression in renal biopsies and peripheral blood leucocytes in patients with AAV, and whether P2X7R synergises with ANCA in the activation of macrophages and neutrophils in vitro. 2. The role of P2X7R in the EAV model. I will (i) detect P2X7R and its downstream pathways and their cellular localization; (ii) use a P2X7R antagonist to prevent or treat disease and measure autoantibody production, inflammasome activation, cytokine production, and severity of renal and lung injury; (iii) the effect of P2X7R genetic deletion in WKY rats will be analysed as in (ii); (iv) use bone marrow transplantation between wildtype and knockout rats to determine if immune or renal cells are more important in mediating disease. 3. The role of P2X7R in antigen presentation and leucocyte responses to ANCA in vitro using dendritic cells, T cells, macrophages and neutrophils from WKY rats using P2X7R knockout and antagonist.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Processing and activation of IL-1 related cytokines in psoriatic inflammation.
Funder: Medical Research Council

Abstract: Psoriasis is one of the most common immune-mediated inflammatory diseases affecting up to 4.8% of Europeans. It affects the skin, joints and in some cases is associated with significant co-morbidities including Crohn's disease and increased risk of cardiovascular disease. The disfiguring nature of skin psoriasis, often leads to multiple detrimental sociopsychological and economic consequences. Importantly, 20% of people suffering from skin psoriasis are affected by psoriatic arthritis. This element of psoriatic disease is debilitating and without early appropriate therapy causes permanent joint destruction similar that observed in rheumatoid arthritis. The complex genetic and environmental etiology of psoriasis leads to heterogeneity in disease subtype, progression and responsiveness to drugs. We therefore require further molecular insight into the initiation and progression of the disease in order to provide new drug targets, and need new diagnostic/prognostic tools to allow the subtyping necessary for early and effective therapeutic intervention. From published and unpublished results showing the dramatic and selective gene regulation, and the functional requirement for post-translational processing, we hypothesise that the regulation and proteolytic activation of IL-1 related cytokines is critical in initiating and modulating psoriatic inflammatory. Our objectives for this proposal are: Using patient biopsies, already generated recombinant proteins and inhibitor screening we will: 1 "Isolate mature IL-1 related cytokines generated in psoriasis and determine the proteases responsible"Using newly generated unique monoclonal antibodies suitable for assaying serum and synovial fluid we will: 2 "Compare IL-1 related protein expression in psoriasis subtypes and other immune-mediated inflammatory diseases"Using a skin equivalence model from healthy/patient-derived cells we will: 3 "Validate the role of IL-1 related cytokines in a 3D skin equivalence model"


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Identification of HLA-B*27 bound peptides involved in the pathogenesis of Ankylosing Spondylitis and Reactive Arthritis
Funder: Medical Research Council

Abstract: Although the link between Ankylosing Spondylitis (AS)/Reactive Arthritis (ReA) and HLA-B*27 has been recognised for 40 years, the pathogenic role played by the disease-associated MHC class I subtypes is not understood. Various theories have been proposed, including models implicating the unusual biology of HLA-B*27 that drive the production of aberrant protein forms which contribute to inflammatory disease. The alternative and original, 'arthritogenic peptide' model suggests that disease pathology relates to normal HLA-B*27 forms, where pathogen-specific HLA-B*27-restricted T cells recruited during natural infection subsequently cross-react with self-peptides bound to HLA-B*27 and promote autoimmunity. Recent GWAS studies have re-ignited interest in this latter model, but the lack of powerful techniques to perform peptide-hunting screens have severely hampered progress in this area. In this study, we want to apply newly developed, cutting-edge MHC class I-peptide yeast display technology to re-appraise the 'arthritogenic peptide' model of disease. For the first time, this approach offers a non-bias platform to hunt for peptides that might be linked with disease. Used in conjunction with disease-associated T cell receptor screening tools, the HLA-B*27 yeast platforms will allow high-throughput screening experiments to be performed. Our goal is that this study should dissect the relevance of this model in relation to disease pathology, whilst also driving discoveries amenable to therapeutic development.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: A pathway regulating neutrophil integrin inactivation and its contribution to inflammation
Funder: Medical Research Council

Abstract: The proposed work explores modulation of the activity status of neutrophil integrins, specifically integrin inactivation, and how this critically regulates neutrophil transendothelial migration, neutrophil recruitment and inflammation. The work builds onto our recent in vitro work, that demonstrated that the GTPase activating protein (GAP) ARAP3 is a negative regulator of beta2 integrin activity and integrin-dependent processes in the neutrophil. The proposed work will analyse primary murine and human neutrophils, as well as (human) PLB-985 cells, which are amenable to lentiviral transduction and can be induced to behave neutrophil-like. Three aims will be addressed. (1) To characterise which neutrophil integrins are regulated by ARAP3 in terms of their activity status, the mechanism by which this occurs (surface availability and/or activity status), and whether this activity is conserved between murine and human neutrophils. (2) To identify how this modulation of integrins affects neutrophil transendothelial migration in vitro, and neutrophil recruitment to sites of sterile inflammation in vivo. (3) To address the effects of ARAP3-dependent regulation of neutrophil integrins on the induction, persistence and resolution of inflammation in models of acute and chronic inflammation. The proposed work will contribute to our understanding of the regulation of neutrophil integrins, analysing their inactivation, a previously underexplored step. This work will furthermore characterise the importance of integrin inactivation to neutrophil recruitment and inflammation. The work may lay the foundations to the development of new therapeutic approaches targeting neutrophil integrin inactivation in chronic inflammation.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Development of novel JAK3 inhibitors for the treatment of autoimmunity.
Funder: Medical Research Council

Abstract: This project will carry out hit-to-lead optimization on a novel JAK3 inhibitor we have identified as a potential lead compound for development to treat autoimmunity. Our compound inhibits JAK3 kinase activity in vitro with an IC50 of 4.8nM in a fixed time-point assay and has at least 180-fold selectivity over other JAK isoforms. It is a covalent inhibitor and exhibits the expected time-dependent inhibition of JAK3 kinase activity. Profiling against extended kinase panels has confirmed its in vitro selectivity. In primary immune cells the compound inhibits signaling by JAK3 dependent cytokines at 500nM but not signaling by cytokines requiring JAK1, JAK2 or Tyk2 independently of JAK3. Preliminary DMPK analysis has found that the lead compound has good aqueous solubility, good metabolic stability in vitro when incubated with human liver microsomes and good oral bioavailability in mice. The project will be divided into 3 sections; hit-to-lead optimization, testing of selected compounds in mouse arthritis models and the development of a chemical genetic model to validate the in vivo on target action of the inhibitors. The project will bring together the groups of Simon Arthur, Kevin Read (Dundee), Nathanael Gray (Boston) and Iain McInnes (Glasgow) to form a consortium that has all the required expertise to support the project. Hit-to-lead chemistry will be carried out by Nathanael Gray, whose lab in Boston identified the lead compound. This will be aided by the availability of a JAK3/lead compound crystal structure and supported by kinase profiling and DMPK analysis carried out in Dundee. Selected compounds will be analyzed for their effect in vivo on Jak3 dependent biomarkers and in mouse arthritis models (Dundee and Glasgow). The chemical genetic mouse model will use a point mutation that does not affect catalytic activity but which blocks inhibitor binding. Once validated, a mouse knockin for this mutation will be generated to provide the chemical model (Dundee).


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: ERA-NET NEURON: Identification and study of different immune cell populations and their role in chronic pain
Funder: Medical Research Council

Abstract: Chronic pain is a major burden on the life of sufferers and society as a whole. While our understanding of the condition is progressing, adequate therapies are still lacking and millions of people are living with persistent, debilitating pain that has no discernable function. One of the most promising developments in recent years has been the recognition that microglial activation is a key driver of neuropathic pain. This unexpected role of neuroinflammation brings us tantalizingly close to potential new drug treatments. However, in order to translate this potential, we need answers to several key questions. First and foremost, which cells are driving chronic pain? The contribution of infiltrating monocytes versus resident microglial cells is unknown. Resolving this issue is a critical first step to target the neuroinflammatory process. A second key question is how exactly do resident microglia and infiltrating monocytes respond in different pain states? How do they mediate pro-inflammatory (and possibly antiinflammatory) cascades? And over what time courses? Are they able to maintain an epigenetic, molecular memory of previous insults, which might then 'prime' them to drive a chronic pain state? The aim of our proposal is to seek answers to these questions. A vital and obligatory step, if we hope to develop novel analgesic strategies directed at spinal neuroinflammatory processes. We are bringing together three laboratories, each with highly-specialised expertise that will enable a unique synergy. In the McMahon laboratory, we will use novel cell sorting techniques as well as highthroughput molecular analyses to define and characterise the different immune cell populations involved. Together with the Svensson laboratory, we will use a clinically relevant animal model to examine the epigenetic profile of resident microglia. And finally, the Svensson and Baron laboratories will use in-depth patient phenotyping and human tissue samples to generate translationally relevant findings. Our work will not only yield a much deeper understanding of the CNS neuroimmune response in chronic pain, but also has the potential to reveal novel upstream mediators and biomarkers that may help identify who will be at risk of developing a chronic pain condition. Crucially, it may therefore bring us one step closer to the elusive goal of adequately treating patients and thus greatly increasing their quality of life.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM