NIHRDataInsights/HRCSData huggingface dataset:
Title: Can bioactive compounds from the diet prevent the onset or slow the progression of osteoarthritis?
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Osteoarthritis (OA) is a degenerative joint disease characterised by degradation of articular cartilage and is a leading cause of disability in the UK. Two major risk factors for OA are increasing age and obesity, thus demographic trends highlight OA as an mounting health and economic problem. There are currently no effective disease-modifying drugs to treat OA and drug development in this area is difficult. A strategy which focuses on bioactive compounds in the habitual diet to prevent onset or slow progression of OA offers a promising alternative strategy. In preliminary studies, we used epidemiology in the UK Twin Registry to identify protective associations between diet and OA. This led us to identify compounds from both allium and cruciferous vegetables which blocked cartilage destruction in our laboratory models. Our hypothesis is that bioactive compounds in the habitual diet have the potential to prevent the onset and/or slow the progression of OA. This proposal aims to extend and refine epidemiology using a co-twin case-control study in the Twins UK database, with more detailed dietary assessment and focusing on OA discordant twins. We will also examine prospective data in the EPIC Norfolk cohort, identifying patients who have progressed to OA ascertained by joint replacement. We will use our earlier laboratory data and these epidemiology studies to test the relative potency of key bioactive compounds from relevant foods in both primary human articular chondrocytes in culture and also in human cartilage explant assays of tissue catabolism. This will culminate in a pilot scale proof-of-principle study in man to examine the levels and functional activity of bioactive metabolites attained in human cartilage after consumption of food containing the bioactive. Ultimately, this will inform the design of a full scale human intervention trial enabling us to translate findings from both an industrial and public health perspective.


Research activity codes: 2.4 SURVEILLANCE AND DISTRIBUTION;3.3 NUTRITION AND CHEMOPREVENTION
Health categories: MUSCULOSKELETAL
Title: Ergocalciferol (D2) vs. Cholecalciferol (D3) Food Fortification: Comparative Efficiency in Raising 25OHD Status & Mechanisms of Action (D2-D3 Study)
Funder: Medical Research Council

Abstract: Growing evidence for the positive effects of vitamin D in reducing risk from disease has increased our awareness of the need for evidence-based strategies to reDr.ess the high prevalence of low vitamin D status in the UK. This requirement has special relevance to a multi-cultural Society in which large numbers of important Minority Groups are at significantly increased risk of hypovitaminosis D. While food fortification is a sustainable solution for the prevention of vitamin D deficiency, the Food Industry needs to determine the most effective means of carriage, optimal concentration and chemical form of vitamin D if it is to maximize the effectiveness of fortification. This RCT proposal will provide much needed evidence that will; (a) directly compare the biological effects of vitamin D2 vs. vitamin D3 in fortified foods of different matrices (i.e. a solid vs. liquid food); (b) use a dose of vitamin D that is realistic and relevant to the Food Industry and the RNI (400IU); (c) examine responses in Caucasians and South Asians; (d) explore 'systems' level (genetic/enzymatic) differences in potential responses to vitamin D fortification. We hypothesise that at least some differences in the responses of participants to vitamin D fortification will be attributable to differences in gene expression. The VDr. is a nuclear transcription factor which controls expression of a wide range of genes, including several that encode cytochrome P450 enzymes and cytokines. Human leukocytes are known to express the VDr.-encoding gene and it is therefore envisaged that differences in vitamin D responses of individuals could be reflected by differences in leukocyte gene expression. We will determine what foods would be the best vehicles for fortification and whether there is a significant difference between the efficiency of fortification with vitamin D2 vs. vitamin D3 in their ability to raise 25OHD levels given that vitamin D intakes are insufficient in the UK throughout the year.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: MUSCULOSKELETAL
Title: Kick-starting mechanoadaptation in aged bones
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Adaptation to loading (mechano-adaptation) maintains bone mass and strength. Relationships between applied loads, the strain patterns generated, and ensuing structural changes remain largely unknown; precise 3D registration and measurement have made it impossible. Three advances now create a unique opportunity to identify the basis of this relationship. We have developed: i) a non-surgical in vivo mouse tibial loading model, ii) a means of 'mapping' strains across entire surfaces of such loaded bones by digital image correlation (DIC), and iii) acquired new technology for 'slice and view' fluorescence imaging and 3D reconstruction of entire bones. We will combine these approaches to address three aims Our pilot DIC studies have mapped strains on normal tibiae and tibiae which have undergone adaptation to in vivo loading (2 weeks), to establish that this had minimised high strain gradients. Our first aim is to examine whether bone formation occurs in regions of high strain gradients. Skeletal fracture vulnerability in ageing is due to a mechano-adaptive failure. Whether aged bones lose responsiveness due to an insufficient mechanical triggering stimulus or inadequate cellular load-sensing or bone synthesis is unclear. Our pilot studies show that mechano-adaptation fails in aged bones (despite induction of generally higher strains), and our second aim will determine whether ageing bone exhibits a reduced mechanical stimulus and/or in cellular sensitivity to strain. Our final pilot data found that mechano-adaptive responses are restored to aged bones by prolonged neurectomy-induced disuse and our third aim is to establish whether the osteogenic activity in aged bone can be 'kick-started' by introducing regions of high strain gradient. This proposal will identify how we can restore adaptive responses to ageing bone and define strain: bone formation relationships allowing the pathways that translate and communicate the effects of loading to be determined.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: MUSCULOSKELETAL
Title: An innate protector of ageing cartilage: FSTL3
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Follistatin-like 3 (FSTL3) is a secreted glycoprotein that binds and inhibits a subset of transforming growth factor beta (TGFb) ligands, including activin. To reveal its physiological roles we generated FSTL3 gene-deleted mice (FSTL3 KO). Emerging evidence pinpoints a crucial role for TGFb ligands, and particularly activin, in the maintenance of articular cartilage. Our pilot studies using FSTL3 KO mice reveal the pathological outcomes of unrestrained activin activity. We found that ageing FSTL3 KO mice suffer a series of severe joint phenotypes underscoring the importance of FSTL3 action in maintaining cartilage health, including: aggressive cartilage lesions, osteophyte formation, meniscal and cruciate ligament deformations and more rapid closure of growth plate upon cessation of growth, compared to age-matched controls. Herein we will test the hypothesis that FSTL3 restrains activin-induced chondrocyte turnover to maintain normal adult cartilage and prevent its progressive loss during ageing. Using tissue-specific, inducible or global knockout mice, in vivo mechanical challenge and cell culture models we will elucidate the roles of FSTL3 in chondrocyte turnover, a key determinant in cartilage maintenance. Specifically, we will investigate whether: 1) the effects of FSTL3 as an innate protector of cartilage are evidenced only after cessation of growth or during repair of cartilage after injury; 2) FSTL3 expression in chondrocytes alone is sufficient for cartilage maintenance after cessation of growth or whether there is a requirement of a systemic FSTL3 effect. We will also discover the signalling pathways and gene transcription events, influenced by FSTL3, that are crucial for maintaining chondrocyte number and homeostasis. Our findings will define the processes underpinning the maintenance of cartilage integrity during adulthood and ageing, and support development of preventative and therapeutic strategies for cartilage disorders such as osteoarthritis.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: OPTIMISING STEM CELL THERAPY: INVESTIGATING CLONAL HETEROGENEITY IN EQUINE CHONDROPROGENITOR CELLS
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The overall aim of this project is to investigate clonal heterogeneity in equine articular cartilage progenitor cells and, in turn, determine cell lines with optimum chondrogenicity. Currently, there is a knowledge gap in how to identify which cells, following clonal isolation and expansion, will go on to form cartilage capable of functional repair. Data from this study will allow us to determine optimal cell lines that provide the greatest potency in terms of chondrogenesis. Understanding heterogeneity has great clinical importance and these data will assist us to determine whether, for therapeutic purposes, a single clone or polyclonal method is the best approach. If single colonies prove to be the most suitable approach, these data will also enable us to determine which cell line may provide the greatest reparative potential. We aim to reduce such heterogeneity in equine chondroprogenitor cells and optimise their therapeutic use by fully investigating clonal variation and determining optimum chondrogenicity through studying gene expression, immunohistochemistry, biochemical analysis and nano-mechanics. We, therefore, wish to focus on 3 main aims: 1) Investigating the importance of single colony morphology 2) To isolate clonal (single clone) and polyclonal (multiple colonies/clones) cell lines and to compare and contrast chondrogenicity in addition to determining the effects of mixing single colonies of varying chondrogenicity 3) Investigating nano-mechanical properties of expanded single colonies. Thus, by further characterisation of these cells in vitro, we can better understand the relationship of cartilage progenitor cells in development, proliferation and repair, with the long term aim of improving the clinical outcome. This work will also bear relevance to the setting up of immortal cell lines for potential allogeneic procedures in equine cartilage repair.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Overcoming the blunted response to resistance exercise with fish oil to maximise the maintenance of muscle mass in older people
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Mechanisms and strategies to counteract sarcopenia are beginning to be uncovered. Recent work has suggested that increased consumption of EPA/DHA increases protein metabolism and muscle hypertrophy. However, these studies have generally been carried out in young healthy animals or cancer patients. Our pilot data in elderly women found that EPA/DHA supplementation results in a greater adaptive response in skeletal muscle contractility in response to resistance exercise, highlighting a beneficial effect compared to placebo. Further work has indicated that increased consumption of EPA/DHA can help preserve lean muscle mass, in ageing rats, and identified the PI3K/PDK1/p70s6k pathway as a potential mechanism of action. What remains to be determined is whether these effects are seen in a larger cohort of males and females and to determine the mechanisms behind these effects. The aims of the current proposal are to investigate the effect of EPA/DHA consumption on skeletal muscle mass, function and anabolic/catabolic signalling during resistance training in the elderly and whether the responses are the same in males and females. These aims will be addressed by determining changes in functional abilities and skeletal muscle contractile properties after a EPA/DHA supplementation and resistance exercise training. The cross sectional area, volume and fat content of skeletal muscle will be assessed by MRI, before and after the intervention, as will systemic levels of inflammatory markers that are implicated in the aetiology of sarcopenia. In muscle samples we will identify changes in signalling protein pathways associated with increases in protein synthesis and decreases in protein breakdown, potentially identifying mechanisms of EPA/DHA action. These studies will establish whether EPA/DHA consumption alongside resistance training can reduce the deleterious effects of sarcopenia and begin to uncover the molecular mechanisms


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: MUSCULOSKELETAL
Title: The regulation of muscle stem cell migration in ageing
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Skeletal muscle contains a resident stem cell population called satellite cells that in the young have a remarkable capacity to generate precursors to facilitate muscle growth, homeostasis and repair. However satellite cells in aged muscle are not as potent and results in aged related muscle loss, a process called sarcopenia. Satellite cells are normally found in a quiescent state and become activated and then move to supplement existing muscle during tissue maintenance or to repair damaged tissue. We have recently shown that aged satellite cells move at half the speed of young cells. Furthermore we have mathematically modelled movement and discovered that young cells migrate with diffusive (memoryless) characteristics whereas old cells move in a super diffusive manner typifying directionality. The speed and migration characteristic of old cells were converted to those of young cells by the action of Nitric Oxide. We will use the results of our previous study as a platform to investigate in detail the relationship between Nitric Oxide and the migration of satellite cells. We will use time lapse video microscopy to determine how satellite cell migration speed change over time to identify when the speed deficit first manifests. Secondly we will use robust novel techniques to determine temporal changes in Nitric Oxide levels in skeletal muscle. Thirdly we will determine the efficacy three of regimes that increase Nitric Oxide signalling (either by direct exogenous provision of Nitric Oxide, increase in endogenous levels of nitric Oxide production or prolongation of intracellular signalling responses) on satellite cell migration speeds of aged satellite cells. Lastly we will use the optimal protocol promoting Nitric Oxide signalling to determine whether it improves whole muscle regeneration.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: The effects of dihydrotestosterone on amino acid transport in ageing mammalian skeletal muscle fibres
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The hypotheses to be tested in this study are that; (1) sarcorpenia (the gradual decline in skeletal muscle mass and strength with age) is caused by the progressive failure in amino acid transport into and out of skeletal muscle fibres especially type 2 fibres. (2) This decline in amino acid transport is due to an age-dependent reduction in the number of amino acid transporters or their reduced/altered sensitivity to stimulation by anabolic factors such as the male sex steroids. To test these hypotheses, we will perform the following investigates; (1) the effects of age on the expression and physiological functions of SNAT2, LAT1 and LAT2 in fast- (type 2) and slow- (type 1) twitch fibres isolated from mice aged between 60 and 730 days. (2) The cellular and molecular mechanisms underlying these effects. (3) The effects of treating type 1 and 2 fibres isolated from mice ~730 days old with DHT on force production, amino acid transporter expression and function as well as the cellular and molecular mechanism underlying these effects. (4) The effects of treating mice >560 days old with DHT for 4 weeks on skeletal muscle mass, strength, the expression and function of SNAT2, LAT1 and LAT2 and protein synthesis. (5) The cellular and molecular mechanisms underlying these effects. The results from these experiments will provide important information on the role a decline in amino acid transport and reduced bioavailability of anabolic steroids, play in sarcopenia. They will also increase our understanding of the changes that occur in skeletal muscle with ageing and the mechanisms underlying these changes. They will also provide information on whether DHT treatment can delay/slow the development of sarcopenia as well as the mechanisms underlying its effects. The findings from this study will have important implications on in the management of sarcopenia in the future.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: MUSCULOSKELETAL
Title: Microfluidic fibre extrusion for bone replacement
Funder: Biotechnology and Biological Sciences Research Council

Abstract: 1. Use microfluidics to extrude polymers: collagen, polycaprolactone (PCL) and polylactic acid (PLA) in isolation and in combination (fibre composites) 2. Fibre composites will be prepared with each combination and permutation of the three polymers in terms of which are present and their location within the fibre 3. Grow mineral crystals (apatite) along with the polymer fibres and monitor crystal maturation in real time using in situ Raman 4. Use scanning electron microscopy (SEM) to quantify the number and location of mineral crystals with each fibre composite 5. Correlate data from 3 back to the precise conditions at any given point along the microfluidic channel 6. Test the material properties and biodegradability of these composites and use this knowledge to refine our selection of reaction conditions to exclude those that are less suitable in terms of physical and material properties or degradability 7. Test the response of mesenchymal stem cells (MSCs) to these composites. Cellular response will be quantified in terms of attachment, spreading and production of bone markers, stem cell markers (stro-1) and other phenotypical markers will be quantified 8. Use the knowledge in 4-7 to exploit the precise control afforded by microfluidics to refine selection of conditions used to produce potential bone therapy materials via this approach


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Defining the post-transcriptional regulatory mechanisms controlling the SOX9 gene and their potential for promoting cartilage regeneration
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The transcription factor SOX9 is a critical regulator of the chondrocyte phenotype and is involved in the regulation of a large number of cartilage extracellular matrix genes. Reduced expression of SOX9 is a hallmark of diseases such as osteoarthritis, which are strongly associated with old age. In addition, it has been demonstrated that elevated levels of SOX9 are able to strongly promote the formation of appropriate extracellular matrix during cartilage tissue engineering. Therefore, it is sensible that promotion of SOX9 expression forms a part of cartilage regeneration approaches. This project builds on the applicant's published and preliminary data to examine the molecular mechanisms that control this important cartilage gene at the post-transcriptional level. Using siRNA knockdown experiments, the RNA binding proteins responsible for promoting SOX9 mRNA decay will be identified and the relationship between their regulation and that of microRNAs that target SOX9 will be determined. AU rich elements within the SOX9 3'UTR that specifically induce destabilisation will be identified using a cell reporter assay and the RNA binding proteins that interact with these elements will be identified using electromobility shift assays. Finally, the findings will be translated into a tissue engineering approach by examining how knockdown of RNA destabilising proteins, identified in the first set of experiments, or expression of 3`UTR decoy molecules, derived from sequences identified in the latter experiments, can promote the chondrocytic differentiation and subsequent cartilage matrix synthesis of adult human stem cells grown in three-dimensional culture systems Overall, the project promises to deliver in two major ways: (1) characterise a key step in the regulation of a critical cartilage gene, which has great relevance to degenerative diseases affecting the elderly and (2) translate the findings into a technique that will promote the regeneration of cartilage tissue.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;5.2 CELLULAR AND GENE THERAPIES
Health categories: MUSCULOSKELETAL
Title: Denervation of small numbers of fibres during ageing leads to dysregulation of ROS and functional changes in whole skeletal muscles
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Ageing causes loss of fibres from skeletal muscle and the fibres that remain show changes that limit muscle function. These include reduced specific force generation, attenuation of adaptive responses to contractions and increased generation of pro-inflammatory cytokines. The loss of fibres is associated with loss of whole motor units that occurs following cycles of denervation and re-innervation of fibres. Whether this fibre denervation affects the function of adjacent fibres has not been examined, but denervation of whole muscles causes large increases in ROS (hydrogen peroxide) generation by mitochondria. The increase is such that if only 1 in 100 fibres were denervated, this could increase apparent hydrogen peroxide generation from mitochondria isolated from whole muscle to the extent which has been claimed to mediate loss of muscle function during ageing. This project will use a novel partial denervation model to examine whether denervation of small numbers of fibres in a muscle leads to age-related changes in function in adjacent innervated fibres through increased mitochondrial ROS generation in the denervated fibres. The project will examine transection of the superior muscular nerve as a model to denervate a defined group of fibres in the mouse medial gastrocnemius. Mitochondrial ROS generation in small bundles of permeablised denervated muscle fibres will be compared with permeblised innervated fibres from this muscle and microdialysis techniques will be used to determine whether partial denervation increases interstitial ROS in the innervated part. Effects of partial denervation on the function of adjacent innervated fibres will be examined by assessing their specific force generation and adaptive responses to an isometric contraction protocol. Finally, whether effects of denervation are caused by hydrogen peroxide (rather than other ROS or cytokines) will be examined by pre-treatment of mice with polyethylene glycol-tagged catalase.


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