NIHRDataInsights/HRCSData huggingface dataset:
Title: Function of CXCR2 signalling within the articular cartilage
Funder: Medical Research Council

Abstract: Osteoarthritis (OA) is characterized by cartilage breakdown and is a leading cause of disability for which we do not have a cure. We discovered that ELR+CXC chemokines retained within cartilage through binding to heparan sulphate proteoglycans (HSPGs) have an unsuspected homeostatic function. Disruption of their receptor CXCR2 predisposed mice to experimental OA with chondrocyte dedifferentiation and apoptosis; similar data were obtained in human chondrocytes by blocking both CXCR1 and 2. CXCL6 supported human chondrocyte differentiation in vitro and, during embryogenesis, was expressed in the future permanent articular cartilage. The scope of this application is to understand the mechanism by which ELR+CXC chemokine signalling protects cartilage in osteoarthritis without causing inflammation, with the final goal of developing a therapeutic application. Aims: 1) To isolate the function of CXCR2 within adult cartilage from developmental or inflammation phenotypes we will induce OA in a cartilage-specific, tamoxifen-inducible CXCR2 knockout mouse. 2) We will compare the cartilage homeostatic potency of different CXCR1/2 ligands. 3) We will study the biochemistry and function of ELR+CXC-HSPGs interactions in the context of cartilage homeostasis by HS digestion, mutagenesis of the HSPG binding residues, and by binding assays. We will identify the chemokine-HSPGs binding partners by co-immunoprecipitation followed by western blotting of candidate HSPG or mass spectrometry. 4) We will test if we can inhibit the capacity of ELR+CXC chemokines to cause inflammation without altering their homeostatic properties by disrupting HSPG binding (necessary for the formation of haptotactic gradients and neutrophil extravasation) and incorporating these mutant molecules in a bioscaffold. The efficacy of this strategy will be tested in an in vivo cartilage formation assay involving the adoptive transplantation of human chondrocytes and neutrophils in nude mice.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: Investigating the role of the RNA binding protein HuR in musculoskeletal development and disease
Funder: Medical Research Council

Abstract: This project will determine the role of the RNA binding protein HuR in the control of skeletal development by focussing on its role in cartilage differentiation. HuR is a post-transcriptional regulator of mRNA and can reduce the turnover of target transcripts in the cytoplasm. We have observed localised loss of HuR expression in critical regulatory regions of developing mouse long bone cartilage. This supports previous mouse knockout studies, which have identified a role for HuR in the differentiation, patterning and mineralisation of the developing skeleton. This project will characterise the detailed mechanisms affected by HuR, using in vivo and in vitro experimental systems. Firstly we will examine development and disease in a newly established cartilage-specific, tamoxifen-inducible HuR knockout mouse. Secondly, we will perform cutting edge characterisation of post-transcriptional and translational gene control during in vitro chondrogenesis. Thirdly, we will determine HuR's role in stem cells systems, which offer ready translatability into bone and cartilage tissue engineering. Characterising the processes affected by HuR during skeletal development will enhance our ability to modulate regenerative medicine strategies through the use of existing small molecule regulators. Furthermore, the work will improve our understanding of how post transcriptional gene regulation affects cartilage, which is timely given the emerging role of this tier of regulation in chronic diseases of the elderly such as osteoarthritis. Overall, this work will identify exciting new mechanisms that could underpin future treatments of debilitating, age-related diseases of the skeleton, which represent an ever-increasing socioeconomic burden.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: Mechanisms of activity-dependent muscle growth and repair
Funder: Medical Research Council

Abstract: Our aim is to understand the role of stem/precursor cell diversity and physical activity or the force it elicits in skeletal muscle growth and the generation of muscle mass. We study somitic muscle growth and regeneration in vivo using zebrafish larvae expressing genetic reporters by 3D time-lapse confocal microscopy using molecular genetics, physiological recordings and manipulations and a variety of molecular analytical approaches. We have developed a short-term quantitative growth assay and shown that muscle contraction is necessary for normal larval muscle growth. We have identified distinct muscle precursor cells that reside in different somite regions, participate differently in muscle growth and repair and express different muscle stem cell markers. We have three Specific Aims: 1. To distinguish the role of force and activity on muscle growth. By using our rapid growth assay that gives high content data of great precision combined with next generation sequencing and genome editing, we will determine the time course, perdurance and molecular nature of the growth stimulus. 2. To determine the function of stem/precursor cell diversity in muscle growth. By following the behaviour of individual cells and cell clones as they contribute to growth after specific cellular and genetic manipulations we will distinguish the contribution and mechanism(s) controlling a) new fibre formation, b) cell fusion to pre-existing fibres and c) increase in fibre volume during muscle growth. 3. To analyse the cellular and molecular control of muscle wound repair. By applying the above approaches, we will determine the similarities and differences between normal muscle growth and reactive regeneration from the same populations of cells. Using our past experience with a wide range of model organisms and human myogenesis, we are engaging with clinicians to refine our studies and explore the significance of our findings in the context of human health.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: Early mortality outcomes and cost-effectiveness of palliative radiotherapy for bone metastases in the English NHS
Funder: Medical Research Council

Abstract: 30-day mortality (30DM) has been suggested as a clinical indicator for the avoidance of harm in palliative radiotherapy within the NHS. However, no national analysis has been carried out and acceptable outcomes are not known. Alongside this is the increasing use of SABR; a significantly more costly strategy. This project aims to determine acceptable outcomes and optimal treatment strategies both in terms of avoidable harm and cost-effectiveness through four workstreams: 1. Analysis of the variation in fractionation patterns and 30DM following conventional palliative radiotherapy across the English NHS using a 2012-13 cohort of linked data from the National Cancer Data Repository (approx. 120,000 episodes). Factors affecting 30DM will be analysed using multiple logistic regression and clinically useful indicators developed, considering use of both crude and adjusted rates for feedback to centres. This will be carried out in the MRC Medical Bioinformatics centre, University of Leeds. 2. Systematic review of the literature for the outcomes of SABR for uncomplicated bone metastases. 3. Development of a decision analytic model to assess the cost-effectiveness of a range of treatment strategies for uncomplicated bone metastases; best supportive care, single fraction and fractionated conventional radiotherapy and SABR. This is likely to use a Markov model with a cycle time of one week, informed using standard methodologies, alongside the outcomes of the first two workstreams, collaborations and a patient focus group to inform the impact of the treatment process near the end-of-life. A threshold sensitivity analysis for survival will be carried out to determine the point at which each strategy becomes cost-effective over its closest rival. A deterministic sensitivity analysis will assess the impact of other parameters on the model. 4. Application of the model outcomes within a regional budget impact tool to assess the impact of changes on both finance and capacity.


Research activity codes: 6.5 RADIOTHERAPY AND OTHER NON-INVASIVE THERAPIES;6.9 RESOURCES AND INFRASTRUCTURE (TREATMENT EVALUATION);8.2 HEALTH AND WELFARE ECONOMICS;8.5 RESOURCES AND INFRASTRUCTURE (HEALTH SERVICES)
Health categories: MUSCULOSKELETAL
Title: Antisense Oligonucleotide Therapy for Neuromuscular Disease
Funder: Medical Research Council

Abstract: The use of antisense oligonucleotides (AONs) to target RNA to treat human neuromuscular disease represents a major paradigm shift in medical science. This is exemplified by progress towards a therapy for Duchenne muscular dystrophy (DMD), a catastrophic neuromuscular disorder arising due to out-of-frame mutations in the dystrophin gene and characterised by progressive muscle wasting and premature death. However, while AON-based therapy for DMD is perhaps the most promising of all current therapeutic approaches, this is a critical time for the field. Major scientific barriers to success exist, related to poor systemic delivery of current generation AONs, sub-optimal efficacy in muscle and negligible efficacy in heart and brain, and lack of detailed mechanistic understanding pertaining to AON cell-targeting, cell-uptake, intracellular trafficking and efficacy in relation to pathophysiological outcomes. In this MRC Programme we will build on the international leadership of the Wood Laboratory and on our strong track record over the last 5 years, during which we have successfully begun to address many of these challenges, developing advanced AON chemistries and providing mechanistic insight into AON function and into the requirements for successful targeting of muscle, heart and brain. We now aim to address the fundamental questions that will enable the full potential of AON targeting of RNA to be successfully exploited. Our central objectives include: understanding the chemical and biological requirements for AON efficacy in the neuromuscular system; establishing effective AON targeting of muscle, heart and brain and understanding causal links between pharamacodynamic efficacy and pathophysiological outcome; and elucidating the molecular mechanisms. Our work will open the door to effectively treating numerous neuromuscular diseases, where AON-based precision medical interventions have the potential to transform health outcomes in these areas of high unmet medical need.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: Pathfinder: How do cartilage injuries heal naturally? An experimental study in humans
Funder: Medical Research Council

Abstract: Cartilage defects play an important role in the pathogenesis of osteoarthritis. This fits the conventional wisdom that cartilage, once injured, does not heal. However, conflicting evidence is emerging from longitudinal MRI studies showing that cartilage defects do sometimes heal naturally in humans. We have observed this ourselves in cell-therapy patients, in whom we create a fresh cartilage defect to harvest chondrocytes for cell-expansion, and invariably find this defect healed at repeat arthroscopy a year later. Such natural cartilage healing in humans is poorly explored and understood. This study aims to fill the current gap in our understanding of mechanisms whereby knee cartilage defects heal naturally in humans. It will do so by fully characterising the natural repair process of articular cartilage, from the early inflammatory response, via the cell proliferation phase to the matrix maturation and remodelling phase. The overall research plan is an experimental study in humans using two patient groups: experimental and control. In the experimental group a fresh chondral defect will be created, whereas patients in the control group will have an established chondral defect. The study will therefore compare natural cartilage healing between an acute and an established defect. The proposed study will build a complete picture of the healing response using an innovative mix of techniques, namely biomechanical assessment (gait analysis and joint stability), imaging (MRI and PET/CT), synovial fluid and plasma biomarkers, arthroscopy, immunohistochemistry, biochemistry and mathematical modelling. Taken together, these techniques will provide the pattern over time of the biomechanical, inflammatory, proliferative/apoptotic, cartilage tissue and bone tissue status during natural cartilage repair.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: Disruption of redox relays in the age-related failure of muscle responses to contractile activity
Funder: Medical Research Council

Abstract: Hydrogen peroxide is a key signalling molecule that is generated by contractions and stimulates adaptations in skeletal muscle fibres, but current theories for the mechanisms involved cannot be sustained in the light of the absolute levels of hydrogen peroxide found in muscle fibres. Recent data have shown that some peroxidases, including peroxiredoxins (Prx) and thioredoxins (Trx) are orders of magnitude more reactive with hydrogen peroxide than other proteins and can transmit oxidising equivalents to other less reactive proteins by disulphide exchange. The proposed project will examine the possibility that Prx and/or Trx in muscle mediate the transfer of oxidising equivalents to signalling proteins that regulate multiple adaptations to contractions and that ageing leads to a disruption of these redox-signalling pathways and a reduced ability to respond to contractile activity. The overall objective of the proposed project is therefore to determine the role of disulphide signalling in the responses of muscle to contractile activity, the role that Prxs and Trxs play in these processes and how they are disrupted during ageing. State-of-the-art techniques will be used to assess the formation of intermolecular disulphides in Prx and Trx in muscle fibres following contractions together with a "trapping mutant" approach to identify partner proteins that form transient intra-molecular disulphides with Prx or Trx as a mechanism to transmit oxidising equivalents to the partner proteins. Adaptations of muscle to exercise involve activation of numerous pathways including the transcription factors NF-kappaB, AP-1 and FOXO3 and the essentiality of Prx and Trx in this activation will be assessed by deletion of these proteins by shRNA. Finally, we will translate these data through examination of key components of those redox relays identified by analysis of muscle biopsies obtained from groups of exercising young and older healthy volunteers.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: The role of BET proteins in the differential regulation of myogenesis: Implications for muscle-wasting disorders.
Funder: Medical Research Council

Abstract: The function of BET proteins has so far not been studied in the context of muscle differentiation, although they have been intensely studies in recent years in the context of cancer. BET proteins are ideal targets for small molecule inhibitors as their internal bormodomain motifs consist of hydrophobic pockets which readily accommodate drug molecules in order to directly disrupt their chromatin binding activity. However, the bromodomain motifs are highly conserved between the BET proteins and therefore small molecules BET inhibitors cannot discriminate between family members. As a result, it is likely that key features of BET protein function have been overlooked. Using these same small molecules inhibitors, we have identified BET proteins as important regulators of myoblast proliferation and differentiation. Using RNAi, we have discovered differential roles for BET proteins Brd3 and Brd4 in the regulation of myogenic differentiation in culture. This finding points to a previously unappreciated facet of fundamental muscle biology, and provides a rationale for therapeutic targeting of Brd3 in order to promote muscle growth in the case of muscle-wasting disorders, or for inducing differentiation in rhabdomyosarcoma cells in order to slow cancer progression. The proposed Programme of work will further characterise BET gene function using inducible knockdown and overexpression systems, primary human and mouse cultures, ChIP-seq using anti-BET antibodies, and BET protein-complex composition/post-translational modification by TiO2 phosphoproteomics. Additionally, we will use genetic conditional knockout models to ablate BET expression in satellite cells and challenge muscle with notexin injection to determine the role of different BET proteins in the regenerative response. Lastly, we will explore oligonucleotide approaches to disrupt Brd3 expression in order to promote muscle growth as a therapeutic strategy for treating muscle-wasting disorders/rhabdomyosarcoma.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Targeting circadian clock mechanisms in the intervertebral disc towards therapies for low back pain
Funder: Medical Research Council

Abstract: Circadian clocks drive ~24 hour rhythms in nearly all aspects of physiology and behavior. Changes of circadian rhythms during ageing underpin many of the age-associated pathologies. Low back pain (LBP) is amongst the most prevalent spinal diseases associated with old age. It causes severe pain and loss of mobility, with currently no long-term cure. Progressive degeneration of the intervertebral disc (IVD) tissue is a major contributing factor. However, how susceptibility to IVD degeneration and LBP increases with age is poorly understood. Our preliminary data demonstrates cell autonomous circadian clocks in mouse intervertebral disc tissue and human disc cells. Significantly, our circadian transcriptome profiling data strongly implicate the circadian clock as a key regulatory mechanism for genes involved in disc tissue homeostasis. Therefore, it is timely to explicitly demonstrate the mechanistic links between the IVD clock and disc tissue homeostasis. Specifically, this research will combine our unique transgenic mouse, ex vivo disc explant and experiments utilising human IVDs to test the following hypotheses: 1) Daily osmotic changes associated with diurnal loading cycle can entrain the IVD circadian rhythm; 2) Disruption to the disc circadian rhythm compromises IVD homeostasis and leads to increased susceptibility to degeneration; 3) Targeting IVD clock mechanisms provides a novel approach to modulate the expression/activity of processes linked to disc degeneration. Key methodologies will include histology, RNAseq, qPCR and Western Blotting analysis to measure rhythmic patterns of gene expression, lentiviral gene delivery, real time bioluminescence monitoring of the dynamics of clock gene reporters, ChIP and functional luciferase assays. Importantly, this new avenue of research holds strong potential to identify new drugs (e.g. clock acting compounds) or improve current treatments (such as timing the delivery of drugs) for IVD degeneration and LBP.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: MUSCULOSKELETAL
Title: The Role of Pls3 in the Pathogenesis of Osteoporosis
Funder: Medical Research Council

Abstract: Osteoporosis is the commonest skeletal disorder, and fragility fractures represent a significant and increasing healthcare burden in our aging society. Many key molecular mechanisms that contribute to osteoporosis were identified in studies of rare human monogenic skeletal disorders. Our International Knockout Mouse Consortium (IKMC) skeletal phenotyping pipeline identified markedly reduced bone mass and strength in adult female Pls3 knockout mice. Moreover, loss-of-function mutations in PLS3 have recently been identified in boys with X-linked juvenile osteoporosis, and a PLS3 polymorphism has also been associated with increased fracture risk in a large population study of post-menopausal women. PLS3 encodes Plastin-3 that binds to filamentous actin, but its function is unknown. I hypothesise that PLS3 is essential for normal bone structure and strength, and that deletion of Pls3 in mice represents an important new disease model whose study will reveal the nature of the role of PLS3 in normal bone physiology. The aims of my proposed research are to determine: 1) The developmental and adult skeletal phenotypes of Pls3 knockout mice 2) The cellular and molecular mechanisms that underlie the development of osteoporosis in these mice. Established advanced skeletal and cellular imaging, cell biology and molecular analysis techniques will be used to determine the skeletal, cellular and molecular consequences of Pls3 deletion. RNAseq analysis will determine the effect of Pls3 deletion on gene expression in specific skeletal cell lineages and facilitate the identification of gene networks and signalling pathways involved in the pathogenesis of bone loss resulting from Plastin-3 deficiency. Historically, study of the critical signalling pathways identified in monogenic skeletal disorders has led to the emergence of new osteoporosis treatments. The proposed project similarly has the potential to identify novel targets for drug development in skeletal disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: The functional role and therapeutic potential of vascular niches in bone
Funder: Medical Research Council

Abstract: The skeleton and the bone endothelium form a functional unit with great relevance in health, ageing and disease. The skeletal vasculature provides nurturing niches for bone and blood-forming stem cells and regulates osteogenesis and hematopoiesis. Thereby, the endothelium plays a key role in maintaining the skeletal health, and a dysregulation of the vasculature is speculated for several bone diseases. Increasing occurrence of bone disorders pose a major burden on people, health systems and the Government. I identified a specialized blood vessel subtype termed 'type H' that generates an active niche for osteoprogenitor cells. The abundance of type H endothelium declines in ageing, and reactivation of type H endothelium in aged mice leads to increased osteoprogenitor numbers and improved bone mass, indicative of their therapeutic potential. My preliminary data suggests that distinct blood vessel subtypes support specific mesenchymal stem/progenitor cell (MSPC) subsets in bone. Here, I will characterize the regenerative niches in bone and will define the role of the endothelium and associated MSPCs in bone repair by using irradiation and fracture models. Further, I will demarcate the role of distinct endothelial cell populations in regulating the MSPCs quiescence and fate determination towards osteogenic versus adipogenic lineages in development and repair. Finally, I will elucidate endothelial-mesenchymal regulatory signalling pathways in MSPC differentiation, using transcript profiling, computational biology, in vitro 3D cultures and genetic mouse models. The study will not only elucidate regenerative niches and regulatory angiocrine signaling networks in bone repair but will it will also break new ground by illustrating the vascular control of MSPC quiescence, fate and differentiation, and may identify novel strategies for managing bone pathologies.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL