NIHRDataInsights/HRCSData huggingface dataset:
Title: Products of lipid oxidation produced by synovial macrophages as triggers of articular inflammation: novel therapeutically relevant approaches to their silencing
Funder: Versus Arthritis

Abstract: Rheumatoid arthritis (RA) is characterized by recruitment of leukocytes, including neutrophils, into the synovium[1]. Neutrophils from RA patients overproduce reactive oxygen species[4] that initiate lipid peroxidation within the joint, including peroxidation of arachidonic acid (AA)[5]. Downstream products of AA encompass (1) enzymatically derived eicosanoids that induce vascular permeability and vasoconstriction, promote leukocyte chemotaxis, enhance adherence to endothelium and transedothelial migration[12,14,15,19,20]; and (2) non-eznymatically derived lipids, such as isoprostanes[6], with less defined function in inflammation. The identity of synovial cell responsible for initial release of eicosanoids and isoprostanes that guide neutrophil entry into the articular space is unknown. Our preliminary data generated by advanced imaging techniques indicate that neutrophil entry may begin at the synovial lining area. This project aims to investigate whether a distinct population of synovial lining macrophages release eicosanoids and isoprostanes and trigger directional neutrophil recruitment in response to a challenge. I also aim to explore strategies of direct interference with the lipid pathways in macrophages via blocking a key enzyme in AA processing (cytosolic phospholipase A2 (cPLA2)[8]) using unique genetic models and intra-articular delivery of nanocarrier supported cPLA2 inhibitors. Finally, I will evaluate whether dietary uptake of re-enforced AA that is oxidation resistant leads to attenuated production of eicosanoids and isoprostanes in the joint and silence inflammation. I will employ a combination of state of the art imaging, biochemical and immunological techniques and unique mouse models to address these aims. The outcome of this project will be a mechanistic insight into lipid mediated macrophage-neutrophil crosstalk and novel therapy design targeting oxidation pathways to silence inflammation.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Investigating the pro-inflammatory effects of synovial B-cell derived recombinant monoclonal antibodies in fibroblast-like synoviocytes of rheumatoid arthritis patients
Funder: Versus Arthritis

Abstract: Up to 50% of rheumatoid arthritis (RA) patients display synovial ectopic lymphoid structures (ELS) supporting B autoreactivity toward locally generated citrullinated and other translationally modified antigens. Recently, screening a large number of recombinant monoclonal antibodies (rmAbs) which I derived from locally differentiated B-cells from RA ELS+ synovium, I identified a subset of antibodies which specifically recognise fibroblast-like-synoviocytes (FLS), suggesting FLS as a cellular source of autoantigens fuelling the local autoimmune response. These rmAbs did not cross-react with the anti-neutrophil-extracellular-traps (anti-NETs) that I previously described. RA-FLS have a crucial role in the RA pathogenesis contributing to local cartilage destruction/inflammation by producing matrix metalloproteinase, direct invasiveness of cartilage/subchondral bone and release of pro-inflammatory cytokines/chemokines. It has been proposed that endogenous toll-like-receptors (TLRs) ligands, particularly TLR4, have an important role in inducing a pro-inflammatory response from RA-FLS. Interestingly, TLR4 might act as receptor for immune complexes containing citrullinated antigens. However, these mechanisms have not been fully investigated. Notably, when incubated with FLS, selected anti-FLS rmAbs displayed pro-inflammatory properties with induction of IL-6/IL-8 release. Recently, I reported that calreticulin is one of the antigenic targets of these anti-FLS rmAbs, while the nature of other FLS-derived autoantigens is still unclear, although my preliminary work suggests that heat-shock protein 60, another protein involved in stress-response and an endogenous TLR4 ligand, is targeted by some of the pro-inflammatory anti-FLS rmAbs. Thus, in this proposal, I aim to: i) fully characterise the stromal-derived autoantigens recognised by anti-FLS RA-rmAbs, ii) elucidate the anti-FLS rmAbs functional role in modulating inflammation and the mechanisms involved in RA-FLS activation in vitro, and iii) study the pathogenicity of anti-FLS rmAbs in driving RA joint inflammation/autoimmunity and destruction in vivo. This study has the potential to lead to the identification of new targets for the development of novel diagnostics/therapeutics for clinical use in RA.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Stromal cells as therapeutic targets in soft tissue disease of the joint
Funder: Versus Arthritis

Abstract: Frozen shoulder is a common cause of shoulder pain without a cure. The condition is a unique example of a localised, self-limiting, chronic inflammatory disease of the joint capsule. Patients are symptomatic for 2-5 years, during which time the disease spontaneously resolves. Animal models do not exist and the disease is poorly understood. Access to patient tissues is vital to inform why the condition ultimately resolves and to identify new therapies to accelerate disease resolution. I identified CD90negfibroblasts and CD163+IL10+Lyve1+ macrophages in capsular tissues from frozen shoulder patients, hypothesising that these resolution-phase fibroblasts (rFB) and macrophages (rMØ) are key stromal subsets concerned with resolving inflammation in frozen shoulder. We have established a clinical study to collect capsular tissues from frozen shoulder patients and non-diseased matched tissues, providing a unique resource to identify the pivotal cellular checkpoints regulating successful resolution of self-limiting inflammation. I will use scRNASeq to characterise the phenotype and function of tissue resident stromal subsets, creating a spatio-temporal cellular atlas of the joint capsule. Lipid metabolomics will identify the distinct resolution circuits active in capsular stromal subsets isolated from frozen shoulder patients, such that resolution circuits can be mapped onto identified stromal subsets. This will inform precision therapeutic targeting of stromal cells, identifying if functionally distinct stromal subsets drive inflammation and resolution, or whether stromal cells exhibit a plastic multifunctional phenotype. I will then test if Specialised Proresolving Mediators (SPMs) target stromal cells from frozen shoulder patients, moderating their inflammatory phenotype. Transfection of patient-derived rFB and rMØ subsets with short hairpin RNA (shRNA) to SPM synthetic enzymes and receptors will confirm causal mechanism, identifying the pivotal cellular checkpoints regulating successful resolution. Access to unique patient tissues and cutting-edge platforms will catalyse a step-change in understanding of disease to identify if new proresolving therapies accelerate resolution of frozen shoulder.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Optimising the functional potential of serpins to block cartilage breakdown: Implications for novel therapeutics for osteoarthritis
Funder: Versus Arthritis

Abstract: The proteolytic destruction of cartilage, in particular type II collagen, is a central and irreversible process that underpins osteoarthritis (OA). Collagenases such as MMP13 are key effectors in this proteolysis but their structural architecture makes selective therapeutic targeting over other MMPs challenging. We and others have demonstrated that serine proteinases are important in regulating collagenase activity and cartilage breakdown. Moreover, serine proteinases are proven drug targets in many other disorders. The largest family of serine proteinase inhibitors are the serpins. Through a unique, covalent mode of inhibition, serpins have evolved to directly target specific proteinases. One serpin, SERPINE2, is highly downregulated in OA compared to control cartilage (padj=1.35x10-38). When added to an ex vivo model of cartilage destruction, SERPINE2, and the highly related SERPINE1, block collagen destruction, whereas serpins with different specificities do not. Their known inhibition of trypsin-like serine proteinases, coupled with collagen release also being sensitive to MMP3 inhibition, suggests they are blocking a serine proteinase which functions as a proMMP3 activator, which in turn is a major activator of proMMP13. Thus, elucidation of this pathway will highlight new, potentially druggable targets. These serpins are abundant in cartilage and yet almost nothing is known of their regulation in the tissue, or indeed how this impacts on OA. There are 3 aims to this proposal, which are complementary but importantly independent. 1) Identify proteinase targets of SERPINE1 and SERPINE2. 2) Determine the key mechanisms by which these serpins are regulated in, and by, the cartilage. 3) Characterise the role of serpins in chondroprotection in an animal model of OA, and assess the feasibility of using serpin biology as a way of blocking cartilage destruction. This project is both novel and timely, and highly feasible. Furthermore, it paves the way for new therapeutic avenues for a major unmet clinical need.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: The role of CD8+ T-cells and type-I IFN in juvenile-SLE and accelerated atherosclerosis
Funder: Versus Arthritis

Abstract: Juvenile-onset systemic lupus erythematosus (JSLE) is an autoimmune disorder characterised by chronic inflammation and pro-inflammatory type-I interferon (IFN) signatures. Patients with JSLE have an increased risk of developing cardiovascular disease (CVD) through atherosclerosis, chronic inflammation of the larger arteries, compared to healthy individuals, a leading cause of patient mortality. JSLE patients also have a more severe disease phenotype, increased CVD-risk and mortality compared to adult-SLE. Investigating early developments of atherosclerosis in these younger patients could allow for early intervention to improve long-term prognosis. No guidelines exist for CVD-risk management in JSLE and it is not possible to predict CVD-risk using traditional biomarkers; the precise mechanism is yet to be elucidated. CD8+ T-cells were recently shown to be extremely abundant in atherosclerotic plaques (>30% of immune cells) and circulating type-I IFN levels are associated with sub-clinical markers of CVD in JSLE. In support, my preliminary data suggests a key role for CD8+ T-cells and type-I IFNs in JSLE-associated atherosclerosis. In world-leading research environments specialised in CVD and adolescent rheumatology, I now aim to: Define how type-I IFN-signalling associated with JSLE influences CD8+ T-cell metabolism/function. Investigate IFN-signatures in JSLE patients with sub-clinical atherosclerosis (collaboration with 2 clinical trial sponsors) and compare matched CD8+ T-cell transcriptomic signatures to experimental models of atherosclerosis (1-year placement at New York University). Investigate therapeutic strategies to mitigate atherosclerosis-associated CD8+ T-cell activation by inhibition of IFN-signalling. This will generate novel mechanistic knowledge regarding CD8+ T-cell-mediated inflammation in atherosclerosis and could identify new predictive biomarkers and therapeutic targets for CVD in young patients with JSLE to improve disease prognosis and quality-of-life, an unmet clinical need. Patients have been involved in the development of this project and these aims will run in parallel with frequent patient engagement events to maximise the translational potential of the research outputs.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Modelling cardiomyopathy in inflammatory arthritis to define mechanisms and guide therapeutic strategies
Funder: Versus Arthritis

Abstract: Cardiovascular comorbidities contribute to around half of all deaths in patients with rheumatoid arthritis (RA), among which non-ischaemic heart failure plays a predominant role. RA patients are particularly susceptible to a type of heart failure with preserved ejection fraction (HFpEF), which is accompanied by diastolic dysfunction. The causes of diastolic dysfunction in RA are unknown and current medications for RA do not produce marked cardioprotective effects. To address this unmet clinical need, I have identified a mouse model of arthritis, K/BxN F1 progeny, which recapitulates the specific diastolic dysfunction reported in RA patients; this is the first mouse model of inflammatory arthritis which shows diastolic dysfunction. Based on my preliminary data, I hypothesise that the observed diastolic dysfunction in arthritic mice is due to 1) cardiac fibrosis/hypertrophy contributed by systemic and local inflammation indicated by elevations in multiple circulating inflammatory cytokines and dysregulated infiltration of cardiac immune cells; and 2) substantial metabolic alterations, with uncoupled glycolysis and mitochondrial oxidative metabolism. I have identified a separate second model of inflammatory arthritis also coupled to diastolic dysfunction: hyper-homocysteinaemic mice subjected to injection of arthritogenic KBN serum. In this fellowship, I will address the following key goals: I will utilise K/BxN F1 progeny to elucidate how cellular/molecular dysregulation contributes to the development of cardiac dysfunction by defining time-courses for immune cell infiltration, glycolysis and mitochondrial oxidative metabolism. I will also test the impact of both established anti-inflammatory as well as innovative pro-resolving therapeutic strategies on cardiac function and joint symptoms in K/BxN F1 mice. I will validate discoveries in a second model of arthritic mice with hyper-homocysteinaemia. Finally, I will access RA patients’ data and blood samples from Barts NHS Bioresource to investigate whether diastolic dysfunction/ HFpEF is associated with cardiac fibrosis, and whether systemic inflammation contributes to cardiac fibrosis and/or dysfunction.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Modelling the cellular causes of HLA-B*27 associated spondyloarthritis with single-cell genomics
Funder: Versus Arthritis

Abstract: 1. Primarily, develop and deliver a searchable Directory to enable researchers to discover and contact multiple human tissue and biosample collections via a unified interface (taking account of existing systems) in order to facilitate sample access. The Centre will be expected to provide an evaluation/metrics of the system, in terms of usability by tissue resources and researchers, effectiveness in providing the ability to locate relevant samples, a demonstration of the benefits of the chosen approach to enhance research use of samples, and other metrics;


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Arthritis Research UK Chair in Rheumatology
Funder: Versus Arthritis

Abstract: Support is sought for a Professorial Salary plus additional costs, commensurate with the seniority and competitive expertise of the proposed recruit, Dr Andrew Yates, from the United States. The consumables requested will provide the requisite computing and information technology and laboratory resources to allow powerful mathematical modelling, and validation of the articular immune response, and will allow the applicant to deliver quickly upon locating to the University. This will offer the best chance of 'quick wins' for Dr Yates, for the arthritis research community in the University and especially for the Charity. Some support is requested to allow dissemination of this expertise, and to allow data transfer and analysis from outwith the University. This will enable ongoing collaborative activities on the part of Dr Yates, especially with Prof Ron Germain at NIH, to mutual benefit, but also will enable initiation of new relationships within the UK arthritis community. This will initially be via the ARUK Centre of Excellence in Rheumatoid Arthritis Pathogenesis, but will roll out thereafter driven by cognate expertise, relevant questions and mutual collaborative enterprise. Our vision here is to create a unique expertise in the UK to the eventual benefit of all in the 'ARUK community'.


Research activity codes: 2.5 RESEARCH DESIGN AND METHODOLOGIES (AETIOLOGY)
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Extracellular matrix mediated control of immune cell recruitment and positioning in health and disease
Funder: Wellcome Trust

Abstract: Recruitment and positioning of immune cells is critical to biology, enabling tissue development and combating of infection. However, unregulated leukocyte recruitment can lead to inflammatory diseases including stroke, atherosclerosis and rheumatoid arthritis that contribute to 70% of global deaths. The key control point in leukocyte recruitment to tissues is their movement from the circulation through the blood vessel wall. A crucial, but largely ignored, component of the blood vessel wall is the glycocalyx, an extracellular matrix barrier primarily composed of glycosaminoglycan (GAG) sugars. I will determine whether: • The endothelial glycocalyx lines blood vessels to control leukocyte migration by masking leukocyte access to endothelial adhesion molecules. • This glycocalyx is physically remodelled by inflammatory proteins (chemokines) to allow leukocyte access to endothelial adhesion molecules and subsequent migration. • The composition of the glycocalyx is tuned to facilitate presentation of certain chemokines and recruitment of specific leukocytes at different sites. This is one example where the extracellular matrix in the immune system has been overlooked. I will establish a unique approach encompassing biochemistry, biophysics and in vivo biology to produce unprecedented insight into the mechanistic role of the glycocalyx and chemokines during inflammation; informing future therapeutics to treat a wide range of diseases.

Lay abstract: The immune system allows our body to kill bacteria and viruses (pathogens) that infect us. A key part of this system is how the cells that kill pathogens (immune cells) arrive where they are needed. Chemokines direct immune cells to sites of infection to enable our bodies to attack pathogens. However, sometimes this can be triggered when there is no infection, producing damage of healthy tissue. This can lead to diseases such as cancer, inflammatory arthritis and atherosclerosis that are significant global problems. Despite this integral role of chemokines, and over 20 years of research, there are no therapies in the clinic that target chemokines during inflammation. Thus, drugs targeted at the specific processes mediated by chemokines are a strong prospect for future therapies. This proposal will identify specific steps in the inflammatory process to target and may highlight novel drugs as candidates to treat inflammatory diseases.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: INFLAMMATORY AND IMMUNE SYSTEM
Title: Understanding immune metabolism on the single-cell and structural levels in the normal and autoimmune germinal centre reaction
Funder: Wellcome Trust

Abstract: There is a growing understanding that metabolic state can profoundly influence the outcome of the immune response, through mechanisms still under exploration. However, how immune metabolism varies at the single-cell level, in time and space, in vivo, and in disease, is almost unknown. The germinal centre (GC) reaction is tightly spatially organised and, critical for the production of high-affinity antibodies. The GC reaction is dysfunctional in lupus, because self-reactive antibody specificities escape deletion. The metabolic demands on B cells undergoing the GC reaction are expected to be considerable. There is evidence that proteins with links to metabolism have important effects on GC B cells. However, there has been no measurement of the metabolome of the GC. I hypothesise that metabolism is organised in space and time in the GC. To understand metabolism on the structural level, I will use imaging mass spectrometry to study normal and autoimmune GCs from patients. To determine dynamic changes, I will use biosensors to image metabolite levels in vivo. I will genetically disrupt key metabolism pathways in the GC in mice, and I will also examine how the glycogen synthase kinase-3 signalling pathway affects B cell responses, and its potential as a therapeutic target.

Lay abstract: The germinal centre (GC) reaction is the process through which our antibodies become more accurate and specialised during the immune response. Sometimes the GC reaction can be dangerous, accidentally altering the target of the antibody to our own tissues. In many autoimmune diseases, including the frequently severe disease lupus, these errors are not corrected and contribute to disease. Recent work has shown that that the fuel immune cells use to generate energy can have great effects on their behavior, and I hypothesize that this is important in the normal and autoimmune GC. I will use the technique of mass spectrometry imaging (MSI) to directly visualize elements of metabolism in the GC. I will also use more conventional methods to measure metabolism, and a variety of models in which metabolism can be disrupted. Overall, the aim is to understand how metabolism might be modified in new treatment approaches to autoimmune disease.

Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: INFLAMMATORY AND IMMUNE SYSTEM