NIHRDataInsights/HRCSData huggingface dataset:
Title: Understanding and Ameliorating perturbed signalling and pathogenesis in FSHD
Funder: Medical Research Council

Abstract: Facioscapulohumeral muscular dystrophy (FSHD) is the third commonest inherited myopathy, characterised by a descending, often asymmetric, dystrophy with initial involvement of facial and proximal upper limb musculature, and later progression to certain lower limb muscles. We have examined gene expression changes during muscle formation in human FSHD cells and used mathematical tools to understand which signalling process are perturbed. We found that pathways that control generation of mitochondria and processes associated with mitochondrial function such as dealing with reactive oxygen species are suppressed. It is well known that FSHD cells are more sensitive to oxidative stress, and a recent clinical trial (clinicaltrials.gov number: NCT01596803) reported that administration of anti-oxidants improved aspects of muscle function in FSHD patients. We have found that inhibiting pathways that control generation of mitochondria in healthy human muscle cells generates myofibres with an atrophic phenotype: characteristic of FSHD muscle fibres. Conversely augmenting the action of these pathways controlling generation of mitochondria in FSHD muscle cells makes them generate myofibres that are larger and indistinguishable from healthy human muscle. This project will investigate how regulation of this pathway controlling generation of mitochondria is perturbed in FSHD and test potential therapeutic strategies to improve its function. Main objectives 1. Determine if drugs that increase mitochondrial biogenesis improve the FSHD phenotype in vivo. 2. Validate the mechanism of perturbed mitochondrial biogenesis in FSHD myoblasts. 3. Decipher the action of drugs that improve mitochondrial biogenesis in ameliorating FSHD. 4. Understand the regulation of mitochondrial biogenesis pathways in FSHD. 5: Determine if mitochondrial biogenesis improves DUX4 induced myopathy in vivo.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: MUSCULOSKELETAL
Title: Modulation of the Wnt signalling in the articular cartilage: is it all about the dose?
Funder: Medical Research Council

Abstract: Despite its strong association with osteoarthritis (OA) development, Wnt-signalling has been an elusive target for the generation of disease modifying drugs. Researchers so far have in fact been unable to untangle the components leading to the anabolic effects of the signalling from the pro-catabolic ones. Our previous and most recent data suggest that a tight balance in the activation levels of two branches of the signalling, the Wnt/beta-catenin and the Wnt/Calcium Calmodulin Kinase (CaMKII) cascades, is required to maintain cartilage homeostasis and avoid OA development. In this grant we aim to investigate the molecular mechanisms activating the pathways and keeping their balance, to understand how cartilage physiology is maintained and how to re-establish it to avoid OA development and/or halt disease progression. More in detail, we will be investigating (i) how the specificity of receptor-ligand interaction is involved in the activation of the Wnt/beta-catenin and of the Wnt/CaMKII pathways (ii) which micro-RNAs are involved in modulating Wnt-led signal transduction, (III) whether the manipulation of ligand/receptor interaction and of the key micro-RNAs can influence the chondrogenetic and pro-regenerative properties of the articular chondrocytes. The project builds up on our preliminary data and toolsets (microarray analysis of Wnt beta-catenin dependent targets in articular chondrocytes and wide expertise in in vitro phenotype analysis of chondrocytes and in vivo models of chondrogenesis) and on ongoing collaborations with world-class pharmacologists and experts in the field of osteoarthritis. The grants aligns with MRC Strategic Aim 1 (Picking research that delivers-Theme 1) and Aim 4 (Supporting Scientists).


Research activity codes: 2.5 RESEARCH DESIGN AND METHODOLOGIES (AETIOLOGY)
Health categories: MUSCULOSKELETAL
Title: Combining iPSC and gene editing with cell therapy to develop the next generation of MSC-based therapeutics to counteract bone fragility in OI.
Funder: Medical Research Council

Abstract: Osteogenesis imperfecta (OI) is characterized by skeletal fragility and compromised osteoblast function. Transplantation of human mesenchymal stem cells (MSC) isolated from amniotic fluid (AFSC) in OI mice improved bone quality, stimulated osteoblast maturation and decreased osteoclast activity. However, primary MSC lose their regenerative potential during in vitro expansion, which raises the issue of standardisation and donor cell stability. An alternative replicable source of human MSC is induced pluripotent stem cells-derived MSC (iMSC). In this project, we propose to test the efficacy of human iMSC to counteract OI bone fragility and also to determine secondary solutions for the treatment of OI (transplantation of iMSC-derived osteoblasts or exosomes or soluble proteins released by iMSC). Our first objective will evaluate and compare the efficacy of iMSC (derived from healthy fibroblast-iPSC, AFSC-iPSC and OI-iPSC following correction of the OI mutation) and AFSC to improve bone mechanical and structural properties following transplantation in oim (recessive) and G610C (dominant) OI neonate mice. We shall next evaluate the in vivo efficacy of AFSC-derived pre-osteoblasts, AFSC-exosomes and AFSC-soluble proteins. Finally, we shall use ex vivo cocultures, imaging and qRT-PCR to investigate the impact of OI mutation on the functionality of human and mouse osteoblasts (i) and determine whether osteoblast and osteoclast functions are modulated by MSC, either through direct cell contact, or via the release of exosomes or soluble proteins (ii). Our long-term goal is to develop the next generation of iMSC-based personalized therapy using patients' own iMSC, their osteoblast-differentiated derivatives or their by-products, to counteract bone loss and ameliorate the quality of life of people affected by skeletal fragility.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: In vitro and in vivo studies of 3D orthopaedic implants with cell-instructive nanotopographies
Funder: Medical Research Council

Abstract: This project is aimed at conducting in vitro and in vivo studies cell/microbiological studies of 3D porous Ti implants with nanostructured surfaces; translating our current understanding of 2D nanotechnology into 3D. We have assembled a multidisciplinary team from universities of Bristol, Glasgow and Cambridge in partnership with clinical and industrial partners to address important scientific and technological questions that will permit us to progress towards preclinical/clinical trials. For example (1) do MSCs and bacterial cells respond similarly to 3D curved nanostructured Ti surfaces on porous implants as they do on 2D nanostructured surfaces? (2) do the osteogenic-antibacterial effects remain if MSCs and bacteria are cultured together on the surfaces? (3) do the nanotopographies remain cell instructive in the dynamic in vivo microenvironment? (4) can we further functionalise the nanotopographies to enhance the synergy of antibacterial properties and osteogenic potential? To this end, we will investigate two 'non-line-of-sight' nanostructuring techniques to generate nanotopographies on 3D porous Ti implants and assess their robustness under simulated surgical conditions. We will use flow-cell microbiology screening, stem cell/bacteria co-culturing and infected animal models to study their response to bacteria and/or stem cells in order to understand their interactions and optimise the cell-instructive surface nanotopographies to develop 3D smart implants for orthopaedics.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: MICA: Moving From Autologous to Allogeneic Cell Therapy: Developing Up-Scale GMP Protocols for Orthopaedics.
Funder: Medical Research Council

Abstract: The aim of the current proposal is to identify the optimal source of healthy, potent allogeneic chondrocytes by assessing adult and juvenile knee cartilage donors and juvenile polydactyly and iliac apophysis cartilaginous tissues. GMP chondrocyte expansion protocols for each will be optimised for the Quantum system, supplementing cultures with pooled human platelet lysate (Stemulate). A 'hybrid' process will be used for expansion, whereby chondrocytes are grown first on tissue culture plastic in order to generate the required cell number for efficient Quantum seeding (5-10M). Depending on growth kinetics data, single donor or pooled donor approaches will be considered. Extensive characterisation of each Quantum product will be carried out using flow cytometry to assess immunoprofiles according to the International Society for Cell and Gene Therapy definition of MSCs, as well as markers indicative of chondrogenic potency (CD44, CD166, CD49c, CD39, CD151, CD271, FGFR3, ROR2 and SOX-9). DNA will be extracted and telomere length will be determined using the TeloTAGGG kit (Roche, UK). The primary outcome for in vitro potency will be cartilage forming capacity using standard pellet culture techniques and assessment of glycosaminoglycan production (modified Bern's histology score). RT-qPCR will be used to quantitate relative expression of chondrogenic genes (Sox-9, collagen type II, aggrecan, frizzled-related protein) and genes indicative of hypertrophy (activin receptor-like kinase 1 and collagen type X). In addition, RNA sequencing of chondrocytes and bone marrow derived mesenchymal stromal cells (BM-MSCs) is planned for in parallel clinical studies, in which global gene expression profiles will be compared to clinical outcome measures (primarily Lysholm knee function score at 15 months). Genes shown to predict clinical efficacy in chondrocyte and MSC therapies will be examined in Quantum products using RT-qPCR.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Use of Mendelian randomisation to examine the role of abnormal hip shape in the development of hip osteoarthritis
Funder: Medical Research Council

Abstract: I aim to establish whether there is a causal relationship between hip shape and hip OA: (i) I will derive a range of hip shape parameters in 100,000 hip DXA scans from UK Biobank (ii) I will ascertain radiographic hip OA in the same DXAs as in (i) (iii) I will describe observational associations between hip shape measures derived in (i), and radiographic OA derived in (ii) and symptomatic OA (iv) I will use MR to determine whether observational associations in (iii) represent a causal pathway between hip shape and hip OA Methods: BoneFinder software, funded by the linked Wellcome AUGMENT grant will automatically plot points around the bony contours of 100,000 hip DXAs in UK Biobank. Having refined this method, I will generate a range of shape parameters previously found to be associated with hip OA, including alpha and CE angles (defining FAI and acetabular dysplasia). I will also apply novel statistical shape models, informed by analysis (as part of my fellowship) of the relationship between 2D and 3D hip shape, derived from hip DXA and CT scans respectively, in 3,500 MrOS participants. A classifier which defines osteophytes, joint space narrowing and with them radiographic OA will be developed in a subset of 1,000 manually graded DXA scans before being applied to all 100,000. Observational associations will be analysed between different hip shape parameters and radiographic OA, self-reported hip pain and risk of hip replacement. Following a GWAS, MR using genetic instruments for the different hip shape parameters will evaluate whether hip shape has a causal effect on hip OA. Opportunities: Finding that altered hip shape leads to hip OA in the wider population will stimulate research into the role of abnormal biomechanical loading in hip OA, and novel prevention strategies aiming to correct this. Identification of hip DXA parameters predictive of hip replacement will provide a basis for using these scans to select those at risk of OA progression.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: Understanding the mechanisms underlying myosinopathies
Funder: Medical Research Council

Abstract: A large number of mutations located in human myosin heavy chains are associated with skeletal myopathies termed myosinopathies. However, their molecular and cellular determinants remain unclear. In this new project, we aim to identify the structural, biochemical and kinetic properties of a commonly occuring human mutations in two skeletal myosin isoforms, MYH7 and MYH2. Our analysis shows that most myosinopathy-causing mutations in these myosins are located in the light meromyosin (LMM) region of myosin. This region of myosin is formed of coiled coil, and known to be important for filament formation. We have identified 10 human LMM mutations known to be pathogenic and leading to muscle phenotypes (weakness) and myosinopathies that we wish to focus on. Based on our preliminary data, we hypothesise that these mutations will compromise filament structure with dramatic consequences on the relaxed and active states of myosin molecules in vivo. We further speculate that these LMM defects will result in sarcomeric and myofibre contractile dysregulation contributing to muscle phenotypes (weakness). To test this, we will define how these mutations specifically alter the various biochemical myosin states by using a combination of Mant-ATP experiments, in vitro motility assays and X-ray diffraction and uncover how these molecular functional alterations affect sarcomere integrity, dynamics and cellular contractility (King's College London). In complimentary studies at the University of Leeds, we will determine how each of these 10 mutations affect the structure and filament forming ability, using circular dichroism, electron microscopy and advanced light microscopy. By achieving these aims, our proposal will link the fundamental impairments to the clinical phenotypes. It will also reveal potential mechanisms necessary for the design of novel treatments.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Muscle RING Finger 1 as a signalling hub for muscle atrophy, mitochondrial dysfunction, and insulin resistance in heart failure and diabetes
Funder: Medical Research Council

Abstract: Skeletal muscle wasting occurs in chronic heart failure (CHF) and diabetes mellitus (DM). Our preliminary data have identified a unique patient cohort with coexistent CHF and DM (i.e. D-HF) whom demonstrate severe muscle wasting, mitochondrial dysfunction, and insulin resistance. Muscle impairments contribute to pulmonary complications, disability, and abnormal metabolism to severely limit quality of life and survival. Therapies to inhibit muscle impairments, therefore, will have major benefits in CHF and DM where prevalence is increasing. Activation of the key muscle-specific E3 ligase, MuRF1, is evident in CHF and DM and linked to fibre atrophy, mitochondrial dysfunction and insulin resistance. We have recently developed, for the first time, an exclusive set of novel compounds capable of blocking MuRF1 activation in vivo, supported by our pilot data in mice where treatment normalized muscle impairments during acute wasting. The present project will combine bench-to-bedside experiments that employ state-of-the-art analytical approaches in order to confirm the contribution of MuRF1 to muscle wasting, mitochondrial dysfunction, and insulin resistance in the chronic diseases of CHF and DM, where 4 experimental groups will be assessed (control, DM, CHF, D-HF). In the first approach, muscle biopsies from patients will be extensively characterized for morphology and function, with human primary cell culture experiments used to assess the impact of MuRF1 blockade on in vitro myotube morphology and mitochondrial function. In the second approach, MuRF1 will be blocked in vivo in mice to determine if this can rescue muscle atrophy, mitochondrial dysfunction, and insulin resistance. Validation of muscle protein homeostasis for anabolic-catabolic signaling in human and mouse will be done to confirm clinical translation between species. Cutting-edge proteomics and metabolomics will also be performed to identify unknown signaling targets modulated via in vivo MuRF1 inhibition.


Research activity codes: 2.5 RESEARCH DESIGN AND METHODOLOGIES (AETIOLOGY)
Health categories: MUSCULOSKELETAL
Title: Bone Toxicity following Pelvic Radiotherapy: understanding, predicting and preventing radiotherapy related insufficiency fractures
Funder: Medical Research Council

Abstract: Radiotherapy Related Insufficiency Fractures (RRIFs) are a common consequence of pelvic radiotherapy. Despite advances in radiotherapy techniques, the prevalence of these fractures is not reducing and they lead to pain, reduced mobility and anxiety. There have been no prospective, interventional or quality of life studies related to management of RIFFs. The pathogenesis of radiotherapy related bone toxicity is not well understood. The aims of this project are (i) to address the currently unmet and large clinical need for evidence based management of RIFFs (ii) to better understand the mechanisms underlying RIFFs, define predictive factors for their development and identify patients who are most likely to benefit from treatment. These aims will be addressed with 3 work packages (WP 1,2,3). The objective of WP 1 is to deliver a randomised controlled clinical trial to determine the feasibility, acceptability and cost effectiveness of a "musculoskeletal health package intervention (including assessment and management of bone density and osteoporotic fracture risk, addressing known risk factors for poor bone health and a prehabilitation package of bespoke exercise provided by the GM prehab4cancer programme) in women undergoing pelvic radiotherapy and inform power calculations and an economic model for a definitive UK multicentre trial. WP1 will also allow prospective data collection on incidence of RRIFs. WP2 will utilise blood samples taken at baseline and radiological imaging from patients in the WP1 trial to investigate potential biomarkers for the development of RIFFs, and includes innovative MS-SWATH proteomic analysis and radiomic techniques. WP3 is designed to deepen understanding of the mechanisms of RRIFs with a longitudinal assessment of bone turnover using ELISA based proteomics during pelvic radiotherapy to determine whether bone resorption is increased initially, thus providing a potential target for anti-resorptive therapy.


Research activity codes: 5.7 PHYSICAL
Health categories: MUSCULOSKELETAL
Title: Are mechanisms of skeletal muscle atrophy simply the reverse of muscle hypertrophy across age? A multi-omics approach
Funder: Medical Research Council

Abstract: Ageing is associated with the loss of muscle mass (sarcopenia), where inducing muscle growth (i.e. hypertrophy) is the key goal of therapeutic interventions that combat muscle loss (i.e. atrophy). Elucidating the mechanisms underlying atrophy and hypertrophy across ages is therefore crucial to maintaining a healthy skeletal muscle mass across the lifecourse, which is essential to preserving locomotory function and whole-body metabolic health. Nonetheless, whether the mechanisms of atrophy are divergent or simply the reverse of hypertrophy remains a fundamental, yet unanswered biological question. This fellowship will detail the functional (e.g. muscle strength), morphological (e.g. muscle fascicle length and cross-sectional area), metabolic and targeted and untargeted molecular responses to atrophic and hypertrophic stimuli in young and older adults. Computational modelling, specifically differential and network analysis, will be applied to omic data sets to determine individual genes/ metabolites that are differentially expressed, biologically-related groups of genes/ metabolites that are co-expressed and the interactions between genes and metabolites (i.e. multi-omics), as a function of age and (lack of) mechanical stimulus (atrophy/ hypertrophy). Key candidate gene targets identified from the bioinformatics analysis will be reverse-translated into human primary skeletal muscle cells to experimentally validate network-predicted mechanisms of atrophy and hypertrophy, as a function of age. This will provide a significant enhancement in the knowledge needed for the development of therapeutic interventions aimed at offsetting muscle atrophy and/ or promoting muscle hypertrophy across the lifecourse.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: New therapeutic avenues in inflammatory arthritis: exploring the role of the adiponectin-PEPITEM pathway
Funder: Medical Research Council

Abstract: Inappropriate accumulation of leukocytes is a shared feature in chronic inflammatory diseases such as rheumatoid arthritis (RA) and psoriatic arthritis (PsA), making this an attractive target for therapeutic intervention. We have identified a novel regulatory pathway, in which a B-cell derived peptide (PEPITEM) limits T-cell trafficking into inflamed tissues. This pathway is impaired in patients with RA, but can be rescued ex vivo with synthetic PEPITEM. We have preliminary data showing that PEPITEM therapy reduces disease severity in murine models of inflammatory arthritis. To date no one has systemically dissected the molecular and cellular responses elicited by the adiponectin-PEPITEM pathway in chronic inflammation, or determined whether functional impairment of the pathway is a shared mechanism driving pathogenesis across chronic inflammatory diseases. In this project, we want to understand the importance of the adiponectin-PEPITEM pathway in the development of RA and PsA, and to dissect the molecular mechanisms underpinning the functionality of the pathway and the use of PEPITEM as a new drug. We will interrogate the functionality of the adiponectin-PEPITEM pathway in patients with RA and PsA to determine when functional impairment of the pathway occurs and whether it is a shared feature of disease pathology. Using murine models of arthritis, we will dissect the cellular and molecular responses to the pathway; how these are altered by functional loss of the pathway; and determine the therapeutic benefit of restoring the pathway in vivo. Understanding the molecular mechanism underpinning the functionality of the adiponectin-PEPITEM pathway will provide important clues in pathology in RA and PsA, and more broadly other chronic inflammatory diseases. Current therapeutic strategies target pathogenic processes, but do not cure disease. Our approach is novel as it aims to restore a regulatory pathway to help switch off inflammation to cure disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL