Developing polymeric coatings for mammalian cells offers the opportunity to tune the chemical and physical environment at the cell surface. This emerging technology has the potential to enable significant advances in the field of cell-based therapies and push forward their clinical application in regenerative medicine. One of the most established methodologies to achieve a polymeric coating around cells relies on the use of materials with an overall positive charge that can interact with the anionic cell surface. However, this approach has been proved to be inadequate for the coating of single cells, as positively charged polymers can disrupt the cell membrane and consequently induce cell death. Similarly, the susceptibility of mammalian cells to mechanical and chemical stress has also limited the covalent conjugation of polymer chains through functionalities already present at the cell surface, hence restricting the chemistry available to achieve a homogeneous and long-lasting cell coating. Nevertheless, recent advances have demonstrated that bio-orthogonal click strategies can be used to introduce exogenous functional groups at the cell surface that can be exploited for polymer conjugation. While these strategies have undoubtedly advanced the field of cell engineering, achieving a homogeneous polymer coating with tuneable properties that is able to control cell behaviour remains an unmet challenge. Importantly, investigating how polymer composition and density of conjugation can be exploited to modulate cell behaviour is essential to fully realise the potential of cell coating strategies in the field of tissue engineering and beyond. This is exactly what this project intends to achieve. By developing robust methodologies for single cell coating, we aim to provide a platform to control cell adhesion with the extracellular matrix and surrounding tissue. This is particularly relevant in tissue regeneration, where cells with regenerative potential (tissue-specific or staminal cells) are injected directly into the target tissue and vasculature to promote tissue growth. Often, cell attachment to the existing tissue is low, mainly as a result of inefficient adhesion of transplanted cells to the surrounding environment, which in turn leads to their programmed death and clearance by the circulatory system. In this project, we will use human liver cells as a model system that will allow us to develop the chemistry and deliver fundamental understanding on the polymer structure, molecular weight, and density of conjugation that are needed to achieve a homogeneous cell coating. This cell type has shown great promise for the development of targeted cell-based therapies for liver regeneration. Using liver as a model, our goal is to develop a versatile coating platform that can be applied to a wide range of cells, advancing the field of cell-based therapies for tissue regeneration. The project represents a priority area for the UK and aligns strongly with the EPSRC's prosperity outcomes (Healthy Nation) and the Healthcare Technologies grand challenges. It also tallies with the United Nations (UN) Sustainable Development Goals, specifically Goal 3: 'Ensure healthy lives and promote well-being for all at all ages.'

Software systems pervade modern life; they control everything from fly-by-wire aircraft and financial transfer systems to ABS breaking systems in cars and cooking modes in microwaves. The ability to understand these complex systems, and to make sure that they behave as expected, is crucial. State machines are a formal, diagrammatic notation that can be used to visualise behaviour of these systems in an accessible way. They can also be used as a basis for several rigorous and automated testing and verification techniques.Currently, state machines have to be designed and maintained by hand. This is an expensive and error-prone task, particularly when the system in question is constantly subject to change. Faced with this challenge, a substantial amount of research has been devoted to solving this problem with automated techniques; to automatically infer state machines of software systems, usually from samples of their behaviour. This has resulted in a multitude of proposed solutions from groups around the world.Although these advances are welcome, they have given rise to an important problem: There is no accepted process by which these techniques can be evaluated and compared against each other. There is no evidence to indicate which technique is better than the others, and why certain techniques excel. This in turn hampers further research in the area.With this project, we will address the above problems by organising an international competition to thoroughly compare and evaluate a diverse range of state machine inference techniques. The competition is especially novel because it will employ a range of techniques to compare the results of different techniques against each other. This will identify (a) which techniques are the most effective ones and (b) shed light on the possible reasons for their effectiveness. It is envisaged that this competition will become a regular event, driving research in the area.

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is an outstanding method of chemical analysis, used extensively in academia and industry to characterise complex samples in 2D/3D. Application areas include materials science, biology, healthcare, energy etc. In the analysis the high-energy 'primary' ion projectile impact on a sample surface, causes ejection of 'secondary' molecular ions which are analysed by a mass spectrometer to provide chemically-rich material characterisation. Scanning the primary beam across the sample provides 2D surface imaging (>100 nm lateral resolution) and by sequentially collecting images while the sample is eroded, 3D sub-surface imaging (>3 nm depth resolution). This unique combination of analytical capabilities means ToF-SIMS is unmatched in its potential to determine, in a single analysis, the composition and detailed distribution of multiple, chemicals in complex samples. Importantly, this technology supports 'discovery mode' research, where the analysis is not biased towards pre-selected, labelled compounds, and therefore leads to hypothesis generation. The analysis is highly-multiplexed and comprehensive - hundreds of species can be potentially detected in a single measurement, limited only by the sensitivity of the process, which here we seek to enhance 100-fold. This proposal addresses critical challenges from next-generation samples demanding greater sensitivity, broader chemical coverage and reliable quantification to address issues including sub-cellular drug localisation and nanoscale molecular materials. It builds on our internationally-leading reputation for innovative ToF-SIMS instrumentation. The characteristics of the primary ion are fundamental in determining impact dynamics at the sample surface and the success of the resulting measurement. The challenge of producing intact secondary molecules from the sample has been largely solved using polyatomic cluster projectiles e.g. C60 and Ar2000 which produce ~100 sputtered molecules per impact. However, only ~0.001-0.1% of these molecules are produced as charged ions, which is necessary for their detection. Clearly there is huge room for improvement in the ionisation efficiency. The principle of projectile-initiated chemical reactions promoting ionisation of sputtered species has recently been firmly established by our work and that of others. We must now build on this knowledge and develop complementary approaches to meet the ionisation challenge and deliver quantitative compositional information. We have assembled a multidisciplinary team of international experts from academia and industry, which is uniquely positioned to pursue this important project. Building on >20 years' experience in innovation of SIMS instrumentation, enabled through EPSRC support and close collaboration with UK Industry, we will develop next-generation reactive ion beams and analytical methodology. This will deliver further transformative gains in performance which are critical to meet future application needs. Our novel results will be framed within the context of emerging theory to understand mechanisms of enhanced ionisation and to underpin the optimisation of projectile parameters. They will stimulate further development of theoretical models of the physical processes underlying SIMS and related techniques. The project is highly-adventurous, providing beyond state-of-the-art analytical capability underpinned with new fundamental understanding. We are ideally placed to exploit this through the interdisciplinary research collaborations at the Manchester Institute of Biotechnology and the Sir Henry Royce Institute for Advanced Materials. The vastly increased quality of data will result in new understanding in a wide range of applications spanning many areas of science and technology.

The ability of organisms to survive and thrive in a changing and increasingly exploited and polluted environment depends on appropriate regulation of energy balance. Growing evidence suggests that exposure to pollutants can alter how fat is stored and used in humans and in other animals. Recent research in humans suggests that many marine pollutants can interfere with the way fat tissue responds to hormonal signals. In particular, pollutants make it difficult to lose weight by switching on pathways that increase fat storage, and this could contribute to problems like diabetes and obesity. Large amounts of persistent organic pollutants (POPs) were made in the early part of the 20th century for use in high capacity electrical conductors and inks, insulators and plastics. Although many POPs are not made anymore because they are very toxic, they are extremely stable and remain in the environment for a long time, ending up in the sea from transport in air and water. Other pollutants, such as phthalates, which are important plasticisers, easily make their way into the sea in run off from urban areas and from marine litter. In particular, POPs do not break down very easily, accumulate in fat and become more concentrated as they are passed up the food chain, ending up in liver and fat. Seals are important top marine predators that have to build up a thick blubber layer while feeding at sea, and then use the fat as a metabolic fuel to keep them alive when they come ashore to breed, moult and rest. They have metabolic similarities to obese and diabetic patients. Their need to rely on fat for fuel and insulation makes them vulnerable to the effects of marine pollutants on the way fat tissue works. The higher the level of POPs in the blubber of seal pups, the lower their chance of survival. Fat tissue like blubber is important for storing fat, releasing it into the bloodstream for use by other parts of the body and producing fat-regulating hormones that control how much fat is stored or used. Recently, the genes of some fat-regulating hormones were shown to be switched on more in blubber of seals from polluted areas in the Baltic Sea than in clean areas in the Arctic, suggesting marine pollutants alter energy balance in seals. However, the mechanisms that control how fat tissue works in seals, and the way marine contaminants interfere with this control, are not well understood. If contaminants can prevent seals from releasing fat from blubber to give them fuel when they are fasting on land, they may have to use more of their protein from muscle tissue instead. This could put them at risk of starvation during moulting, breeding and development, even when they are fat. We will investigate whether pollutants alter fat storage and mobilisation in young grey seals, which are most at risk. We will take small blubber samples from live feeding and fasting seals, without harming them, and treat the blubber with pollutants and fat regulating hormones. We will measure levels of genes and hormones involved in energy balance, the ability of blubber to release fat, and its metabolic rate. By comparing these measures between treatments we will begin to find out how energy balance is normally regulated in seals, how it is altered by marine contaminants, and whether seals are more vulnerable during feeding or fasting. This will help predict the effects of pollutants on seal population size, by contributing a better understanding of how contaminants affect survival of young seals and change their energetic requirements. Because seals naturally experience extreme changes in fat mass, have metabolic similarities to diabetes and obesity, and eat fish, like people do, this work will also inform the likely impacts of POPs and phthalates on human fat regulation. This work has far reaching consequences for health and survivorship in seals and other animals, but also for the management of obesity, diabetes and related metabolic abnormalities in humans.

In the UK, musculoskeletal disorders (joint and back problems) affect one in five people long term. While joint replacements are successful, they are challenged by demands of an active and younger population presenting with disorders due to trauma, obesity, or other lifestyle choices. One of the causes for joint and back pain is the deterioration of the different soft tissues acting as cushions in the joints. New surgical interventions are being developed to repair or locally replace those soft tissues in order to delay or prevent a total joint replacement. There is no clear indication yet on which patients benefit the most from them. There is an urgent need to define the type of patients for which new and existing interventions are most beneficial. The local anatomy or level of tissue deterioration differs greatly between patients, and there is currently a lack of biomechanical evidence that takes into account these large variations to help matching patients to interventions. To tackle these issues, this Fellowship, MSKDamage, will develop novel testing methods and tools combining laboratory simulation with computer modelling and imaging. MSKDamage methods will be used to predict the variation in the mechanical performance of a series of treatments at various levels of joint deterioration. This will enable the different interventions to be matched to different patient's characteristics. I will focus on three musculoskeletal disorders and associated repairs: 1. Emerging treatments involving the injection of biomaterials in the intervertebral disc: I will produce realistic testing conditions that can be personalised to a specific patient, assessing each patient's chance of success and identifying areas for treatment optimisation. 2. Evaluation of meniscus repairs in the knee and their interaction with cartilage defects: I will provide new information on the type of cartilage defect that reduces the chances of success of a meniscus replacement in the knee. The research will develop guidance on the type of cartilage defects that need repair for a meniscus replacement to be successful. 3. Optimisation of custom wrist repair: I will help optimise patient-specific wrist repairs so that they reduce the damage in tendons and ligaments in the wrist. MSKDamage builds on my strong track-record in the field and network of industry, clinical and academic collaborators, as well as my recent work that demonstrates the specific information which need to be included in models of degraded tissues in the spine and the knee. MSKDamage aims to (1) develop a methodology to test interventions for a specific patient and its specific tissue degradation, (2) carry out a series of case studies which demonstrate the capacity to test a range tissues disorders and repairs. This work is a particularly suitable for a Fellowship, as it will allow me to develop fundamental engineering tools and methods while engaging with end users for significant economic and societal impact. I will also develop as a leader in the field, leading a growing research group and taking actions for the research community, directly related to the research, with advocacy on sharing more research outcomes openly for creation of more impact, and indirectly related to act as an ambassador for public and patient involvement for research related to computer simulations in healthcare.