NIHRDataInsights/HRCSData huggingface dataset:
Title: The Role of Adipolin in Cutaneous Fibroproliferative Disease
Funder: Medical Research Council

Abstract: Background: Keloids are benign fibroproliferative tumours resulting from skin injury. They cause considerable morbidity and treatment is suboptimal. We have identified a novel homozygous loss-of-function mutation in the C1QTNF12 gene encoding adipolin, in an autosomal recessive family with striking keloids and hypothesise that this single genetic aberration has contributed to the fibroproliferative process. Adipolin is an adipokine that regulates glucose homeostasis and inflammatory responses. Aim: To perform a functional study of adipolin and characterize the state of glucose metabolism in the context of fibroproliferative disease. Objectives: 1. Determine the function of adipolin in dermal fibroblasts 2. Determine the alterations to glucose metabolism in pro-fibrotic fibroblasts 3. Identify and validate targets to revert the fibrotic/fibroproliferative phenotype Methods: Adipolin-knockdown fibroblasts will be treated with and without recombinant adipolin and assayed for insulin signalling, proliferation, extracellular matrix production and signalling pathways associated with fibrosis using both candidate and exploratory approaches. Alterations to glucose metabolism in these cells will be determined by assaying glucose uptake and glycolysis. Comparisons will be made against wild-type keloid fibroblasts as prototypic pro-fibrotic cells. Targets will be identified for therapeutic manipulation from the aforementioned objectives and small molecule inhibitors/activators will be employed to reverse the fibroproliferative phenotype. Scientific and medical opportunities: Understanding the functional consequences of adipolin loss-of-function on the fibroblast and its metabolism will offer new insight and may lead to the discovery of novel approaches to managing disorders of fibroproliferation including and beyond cutaneous scarring.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: SKIN
Title: iRhom2 and the keratinocyte stress response in health and disease
Funder: Medical Research Council

Abstract: Our hypothesis is that iRhom2 is the key mediator in epidermal hyperproliferation and inflammation. This includes its regulation of ADAM17 shedding of inflammatory cytokines and growth factors. Recently, we have also identified iRhom2 as a key co-ordinator of the cytoskeletal stress response via its novel interaction with Keratin 16 and new published data also showing that p63 regulates iRhom2 in a cell context manner. Thus the iRhom2 pathway controls many key aspects of the stress response to both hyperproliferative and inflammatory skin disease and trauma. This proposal will dissect these multiple roles of iRhom2 in normal and disease epidermal biology. We have both human (in vitro) and mouse (in vivo) experimental models to specifically investigate the targeting of the iRhom2 pathway towards therapeutic management of keratodermas and common inflammatory disease including psoriasis.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: SKIN
Title: Characterisation and acceleration of keratinocyte stem cell culture - Rho-associated kinase inhibitor treatment and epithelial sheet gene therapy
Funder: Medical Research Council

Abstract: We propose to use the Rho-associated kinase inhibitor (ROCKi) to efficiently expand keratinocyte stem cells (KSCs) in vitro. Increased KSCs would then produce sufficient numbers of keratinocytes to accelerate the manufactory of epithelial sheet. To determine the potential of ROCKi treated keratinocytes including KSCs for clinical application, we will examine i) the optimal ROCKi exposure time in the culture; ii) the characteristics of KSCs in ROCKi treated cultures; and iii) whether acceleration of sheet culture is achievable. Holoclone formation assay will be used to determine KSCs expansion efficiency. The characteristics of KSCs, in particular their clonogenicity and fate will also be analysed. Clonogenicity of KSCs with and without ROCKi treatment will be assessed in cell model in vitro and in a human:murine chimera skin graft model in vivo. The fate of KSCs in non-treated and ROCKi treated cultures will be examined at single cell transcriptome resolution. Transcriptome changes will be examined in thousands of cells under different conditions (non-treated and ROCKi treated) and culture times (primary and passaged cells). Unbiased cell population separation for these cell cultures will be performed using changes in the cells transcriptomes. Finally, we will test the potential acceleration of ROCKi treated cultures based on the criteria used in our previous phase I trial. Thus, we will assess the suitability of ROCKi treated cells for clinical application from the evidence gathered in this study.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: SKIN
Title: Functional and molecular diversity of skin fibroblasts in systemic sclerosis.
Funder: Medical Research Council

Abstract: Systemic sclerosis (SSc) has high unmet medical need and only limited therapeutic options. Fibroblasts have a pivotal role in skin fibrosis in SSc. Explant cultures from early stage diffuse SSc show a marked profibrotic phenotype, and gene expression profiles are dominated by a TGFb signature. Recent therapeutic trials have reversed this phenotype in explant cells, but the molecular response has not always been reflected by clinical improvement in skin, suggesting that other fibroblast subpopulations may be important. We recently identified a second population of fibroblasts which remain in the skin after outgrowth of explant cells. This late resident population is profibrotic but much less motile or contractile and, we hypothesise, highly relevant to pathogenesis of SSc. Our proposal will define subpopulations of SSc fibroblasts using molecular techniques for single cell analysis of gene and protein expression in skin biopsies of well characterised SSc cases. Our aims are (1) to define differences in functional properties and gene expression between early explant and late resident fibroblasts including bulk RNA sequencing, providing a functional anchor for the remainder of this project. (2) To analyse fibroblast subpopulations from whole skin using scRNAseq utilising biopsies from different stages of disease (early and late), and different antibody status (ATA and ARA) (n=16) and healthy controls (n=4). Creation of t-SNE plots will help identify clusters of cells, and key gene and protein expressions to differentiate them. (3) To further characterise the skin biopsies by identifying where the different subpopulations of fibroblasts are localised in using confocal image analysis. This project will use approaches already established in rheumatoid arthritis and healthy skin to identify subpopulations of fibroblasts, and their discriminating markers. This will define relevance to SSc and potentially identify new therapeutic targets for this challenging disease.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: SKIN
Title: Understanding the Regulation of Epidermal Differentiation Genes by Filaggrin in Eczema
Funder: Medical Research Council

Abstract: Mutations in the FLG gene which codes for the protein filaggrin are associated with the inherited disease ichthyosis vulgaris and are a major predisposing factor in eczema. FLG is a member of over 60 related genes on chromosome 1q21 called the epidermal differentiation complex, which contains genes that are critical to the barrier function of the skin. It is surprising that only FLG mutations are implicated in eczema, while none of the other genes in the EDC, including the related S100 fused genes are not. To investigate this paradox further we knocked down FLG expression in keratinocytes, and found that there was widespread reduction in EDC gene expression, including key barrier proteins such as loricrin, as well as reduction in key structural keratins 1,2 and 10. We found that BMP signalling proteins were increased in expression. I hypothesise that FLG knockdown leads to reduction in EDC gene expression and this exacerbates the barrier dysfunction in eczema, and that restoring EDC and keratin expression, even without restoring filaggrin, would restore or improve epidermal barrier function. I will examine patient skin scale for levels of EDC genes and keratins 1,2 and 10 by a combination of western blot, mass spectrometry and qPCR and correlate this to filaggrin mutation status and filaggrin protein level to identify a disease endotype in which reduction of EDC and keratin genes occurs. I will then look at BMP signalling in our FLG knockdown keratinocytes and then determine whether restoring filaggrin levels by rapamycin treatment, or restoring normal BMP signalling can rescue both EDC and keratin expression and consequently restore epidermal barrier function. This work should give insights into how the skin barrier is regulated in eczema and understanding how to modulate the skin barrier should lead to better treatments.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: SKIN
Title: Exploiting growth factor modulation of alarmin release in atopic dermatitis
Funder: Medical Research Council

Abstract: Atopic dermatitis (AD) is initiated by keratinocytes releasing IL-33 that initiates a type-2 immune response. Our novel findings show that two cellular pathways are intertwined and essential for triggering this immune response both in vitro and in vivo. We have demonstrated that IL-33 is critical both in human keratinocytes and a mouse model of AD. Additional pilot data from our lab demonstrates that the S. aureus-induced IL-33 release from live cells is regulated by EGF. Using two well established models (human and mouse) we aim to unravel the intricate interaction between IL-33 release and the EGF pathway in response to two common environmental triggers S. aureus and house dust mite (HDM). Cellular work will use phospho-arrays to identify key pathway components, which will be studied by western blot and imaging. We will examine IL-33 binding partner candidates within these pathways using immunoprecipitation to identify additional therapeutic candidates for inhibition of IL-33 release (e.g. histone 2B, exportin-1, HMGB1). Candidate inhibitors (e.g. EGF, neutralising antibodies, EGF pathway inhibitors (mTOR, PIK) will be screened after keratinocyte co-culture with S aureus and HDM. We will also evaluate activity of repurposing drugs as novel therapies which have activity in these cellular pathways and are already used in clinic practice. Candidate inhibitors will then be tested in two separate mouse models of human atopic dermatitis: flaky tail (UK) which carry a functional deficiency in a skin barrier protein Filaggrin, a protein consistently shown associated with human AD; NC/Tnd mice (Tokyo) a spontaneous model of eczema. Clinical eczema skin scores, skin barrier function, and immune cell infiltration will be measured after triggering of AD using S. aureus and HDM. Manipulating IL-33 and EGF pathways should aid in development of novel eczema therapies, as well as understanding the pathophysiology of other allergic phenotypes where IL-33 is a key player.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;5.1 PHARMACEUTICALS
Health categories: SKIN
Title: Living with vulval lichen sclerosus: A mixed methods study
Funder: Medical Research Council

Abstract: Lichen sclerosus (LS) is a chronic skin condition which affects the skin around the genitals. This project is about vulval LS, which affects the skin around the outside of the vagina. LS causes intense itching and small painful cuts in the skin, and people with LS may feel embarrassed and alone. It can cause problems for normal activities such as going to the toilet, sitting or walking, and having sexual intercourse. LS is a lifelong condition - there is no cure. Treatment with steroid ointment usually helps reduce symptoms, but does not work for everyone. Some women may need surgery if they have a lot of scarring. There is very little research about vulval LS. In 2018, researchers, patients, and health professionals determined the top priorities for LS research. They put 'Understanding the impact of LS on quality of life' in the top ten. In this study we will explore what it is like to have vulval LS, how people usually access diagnosis and care, and how it affects their quality of life. We will also explore the views of women with LS of issues related to future clinical trials on the topic. This is because in order to answer many of the priorities identified in 2018, clinical trials will be needed. However, we do not know whether women would take part, and what would be important to them in a clinical trial. We will invite women with LS to take part in an interview about their experiences and views. We will give people the option of a telephone interview if they do not want to speak to a researcher face-to-face. We will invite women to focus groups to explore feelings about the current recommended treatment (putting steroid ointment on the affected area). We will use the results from all of this to design a questionnaire for women with LS to answer. This means we will be able to ask lots of people the same questions, and that people who did not want to take part in an interview will be able to tell us about their experience. Questions will be about what areas of their life are affected by LS and how they accessed diagnosis and treatment. We will also work with an artist to create artwork based on the research findings. At the end of the study, we will hold an event at which we display this artwork and hold a discussion between people with LS and healthcare professionals. Four women with LS have joined a 'lay advisory group'. This group will meet several times during the research to give input into the study design, such as designing the interview questions, and helping us share the findings.


Research activity codes: 7.1 INDIVIDUAL CARE NEEDS;8.1 ORGANISATION AND DELIVERY OF SERVICES
Health categories: SKIN
Title: Role of CD1a-reactive fungal lipid-specific T cells in atopic cutaneous inflammation
Funder: Medical Research Council

Abstract: Atopic dermatitis is a common inflammatory skin disease. Reactivity to fungi may contribute to the inflammatory response. The principal aim of the study is to investigate the role of fungal-lipid specific CD1a-reactive T cells in the pathogenesis of atopic dermatitis (AD). Preliminary data suggest that fungal lipids contribute to inflammation in a CD1a-dependent manner, but the underlying mechanisms are unclear. I will collect circulating T cells from AD patients and healthy controls and co-incubate with K562 cells transfected with CD1a or controls which have been pulsed with fungal extracts (K562 are HLAlow so minimise alloreactivity). CD1a-reactivity will be assessed using ELISpot. I will also collect lesional skin samples from AD patients and healthy controls and co-incubate the isolated T cells with K562-CD1a cells pulsed with fungal extracts. In addition, I will use skin suction blisters raised after injecting commercial extracts of Malassezia spp., C. albicans, or control; these are routinely available for allergen skin testing. T cells will be investigated ex vivo and also expanded in vitro. I will elute lipids from CD1a of the K562 cells pulsed with fungal extracts and then extract and identify lipids isolated from the AD and healthy skin biopsies using liquid chromatography-mass spectrometry in collaboration with Branch Moody (Harvard). Blood and skin CD1a-reactive T cells will undergo single-cell RNA-seq using 10x Genomics to obtain TCR and transcriptomic signatures. I will generate T cell clones using the fungal lipids identified above, and then perform CD1a plate-bound assays to assess lipid loading in a cell free system. In collaboration with Jamie Rossjohn (Monash), I will support the generation of crystal structures of fungal lipid-CD1a-TCR to assess the structural basis for T cell recognition. By improving our understanding of the pathogenesis of atopic dermatitis and mechanisms of fungal defence, we hope to identify novel therapeutic targets.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: SKIN
Title: Genetic and transcriptomic analyses to determine the aetiopathogenesis of solar urticaria
Funder: Medical Research Council

Abstract: Background: Solar urticaria (SU) is a rare, poorly understood skin disease in which normal sunlight triggers mast cell activation and the development of cutaneous wheals. Current evidence supports an underlying genetic architecture that is polygenic. It is frequently associated with atopic disorders and previous experiments suggest that IgE-crosslinking on the mast cell surface by sunlight-activated protein chromophores is a major pathogenic mechanism. However, its epidemiological association with other diseases and evidence of mast cell suppression following ultraviolet irradiation suggest that other modes of mast cell activation and the cutaneous microenvironment may play a role. Aim: To determine the aetiopathogenesis of SU using genetic and transcriptomic analyses Objectives: 1. Investigate shared genetic aetiology of SU and associated disorders using polygenic risk scores 2. Undertake transcriptomic analysis of cutaneous gene expression in SU and healthy controls 3. Validate and explore implicated gene products and molecular pathways Methods: Genetic aetiology shared by diseases can imply common disease mechanisms. Consequently, polygenic risk scores (PRS) will be generated for atopic disorders and other conditions associated with SU. The association of these PRSs with SU will be compared to ancestry-matched healthy controls. Gene expression in lesional solar-irradiated and control non-irradiated skin biopsies from SU patients will be characterized using bulk RNA-Seq and compared with site-matched solar-irradiated and non-irradiated biopsies from healthy controls. Validation will depend on previous findings, but may include immunohistochemistry/immunofluorescent staining of skin or functional interrogation of human mast cell lines. Medical and Scientific Impact: This research has great potential to improve understanding of SU and related urticarias. Such insights may profoundly influence the development of novel therapeutic strategies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT
Health categories: SKIN
Title: Development of Artificial Intelligence OCT Biomarkers for Accelerated Skin Disease Research and Diagnosis
Funder: Medical Research Council

Abstract: The research aims to use AI techniques to develop novel biomarkers for OCT skin scans that will aid end-user interpretation of the scans thus providing added value and utility for the scanning devices. Drawing on an existing library of over 500 scans, including detailed clinical reports, we will use image processing and machine learning techniques to train systems to identify image biomarkers. These will be presented to end-users in a form that will facilitate their interpretation of the scans and allow them to make better informed diagnosis, disease monitoring or treatment planning. The project will deliver working prototypes of AI systems that identify and display 4 novel image biomarkers for OCT scans of skin together with detailed performance evaluation data and release and maintenance plans suitable for their integration into a medical device. Phase 1 of the project plan involves detailed analysis of the technical challenges; understanding the end-user requirements to ensure the proposed solution is relevant; appreciation of background IP to benefit from previously developed and published solutions. Phases 2 & 3 employ rapid prototyping techniques, common for AI system development, to provide early evidence of component efficacy and a regular stream of usable deliverables. Examples of machine learning methods which could be deployed are deep learning classification to identify parts of the scan that are indicative of particular disease or 3D active shape models to delineate structures of interest. The prototypes are useful for internal evaluation and demonstration to customers. Included in the plan, but not costed as part of this project, are work packages assigned to Michelson to provide datasets of images and annotations necessary for the training and evaluation of the AI system. Phase 4 focusses on delivering detailed evidence on performance and efficacy of the prototype systems and a plan for integration with existing Michelson medical device systems.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: SKIN
Title: Skin Metrology for Optimised Automation of the Ten Bio Device Portfolio
Funder: Medical Research Council

Abstract: The individual elements of the work programme will involve: 1: Consultation with the i4PD product development and design team at length in order to assess the most likely next generation architecture for the automated platform and obtain training on the Ten Bio instruments and methods. 2: Prototype development: exploiting the existing instrument platform and architecture at the company with modified electronics and control to facilitate fast cycling and accurate displacement sensing. -> Preliminary measurements. 3. Explore alternative means whereby tension can be assessed/inferred using optical probing as opposed to the contact probe approach presently used, which hampers access in the instrument space. 4. Deploy intense white light source:- an electronically controlled bright LED focussed to the target skin tissue samples and with exposure [period] and duty cycle under full computer control to assess reflective and transmissive components and calibrate with parallel strain gauge based measurements. 5. Examine filter options and the use of polarising plates to assess depth profiling. Reflected light will allow a facility to image the skin surface and assess this for wrinkling signatures that could inform machine vision style control and adjustment to the target tension regime. 6. In assessing risks: scope to move to a more sophisticated measurement using optical coherence tomography if needs be. 7. Implement temperature dependent strain rates measurements by addition of regulated enclosure around the instrument-skin interface. 8. An optimisation approach based on the reliable and widely accepted, but computationally inexpensive Ogden model and analysis will facilitate development of a predictive capacity to assist in the ordering of the manufacturing steps. 9. Generation of new IP covering the extended multi-sample capacity concept. These improvements/supporting data will then be developed for journal dissemination and reported at SID.


Research activity codes: 5.3 MEDICAL DEVICES
Health categories: SKIN