bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Artemis Accords and Subsequent Developments
Authors: de Zwart Melissa , Lisk Joel 
Published: May 2024

Abstract
NASA announced the formation of the Artemis Accords in 2020 with eight foundation partners: Australia, Canada, Italy, Japan, Luxembourg, the United Arab Emirates, the United Kingdom and the United States. As of August 2023, this number has grown to 28. The Artemis Accords provide a blueprint for engagement by these countries on a bilateral basis with NASA on projects which involve the creation of a Lunar Gateway, the return of humans to the Moon and, ultimately, on to Mars. The first Artemis launch occurred in 2022. The content of the Artemis Accords to which a partner agency must agree are expressed to ‘reinforce and implement the Outer Space Treaty, the Registration Convention, the Agreement on the Rescue and Return of Astronauts’ and ‘other norms of behaviour that NASA and its partners have supported’. However, some of the Artemis principles extend beyond these fundamentals of space law to embrace concepts such as safety zones, resource extraction and use and interoperability. These provisions provide challenges to which evolving international space law must respond. This chapter examines the process and provisions of the US-led Artemis Accords, looking at the content and operation of the individual principles contained therein. It also considers whether the Artemis principles represent a unilateral challenge to existing international space law and concludes that, whilst activities undertaken pursuant to the Artemis projects will inevitably create new norms and behaviours, and, in time, laws, the Artemis principles reflect an evolution which will be necessary as humans expand their activities beyond Earth.
O02 Ubiquitomics of CYLD-deficient skin tumours reveals dysregulation of hair follicle keratinocyte processes
Authors: Inns Joseph , Frey Andrew , Rajan Neil 
Published: May 2024

Abstract

Introduction and aims
CYLD cutaneous syndrome (CCS) is an inherited hair follicle tumour syndrome where biallelic loss of function of the deubiquitinase enzyme CYLD gives rise to cylindroma skin tumours. The role of CYLD in skin biology is not well characterized but it is known to regulate several cell signalling pathways through deubiquitination of effector proteins. We aimed to determine how loss of CYLD function can lead to CCS skin tumour formation by studying the ubiquitome of CCS tumours and healthy skin.

Methods
Healthy skin (n = 5) and CCS tumour tissue (n = 5) were collected from flash frozen samples. Lysed homogenates were trypsin-digested and the resulting peptides underwent diglycine immunopurification for subsequent liquid chromatography–tandem mass spectrometry analysis on a Bruker Tims-TOF HT (Billerica, MA, USA) mass spectrometer. We utilized a data-independent acquisition method adapted for diglycine-containing peptides to generate mass spectrometry data. Peptide identification was performed by searching mass spectrometry data against an in silico neural network derived spectral library with diglycine modification.

Results
We identified 1273 ubiquitination sites across 649 proteins in CCS tumour and control samples. After performing t-tests (P &lt; 0.05), we found 305 ubiquitination sites that were either unique to their respective condition or were differentially regulated. Analysis of ubiquitinated peptide sequences uncovered cell surface receptors with increased ubiquitination in CCS tumour samples including the insulin receptor, which had ubiquitination modifications at its kinase domain, suggesting increased insulin signalling in CCS tumours. Gene set enrichment analysis of CCS tumour ubiquitinated substrates revealed increased ubiquitination of secretory granules and extracellular exosomes, suggesting that loss of CYLD may disrupt regulation of protein secretion processes.

Conclusions
Our findings describe the ubiquitome of CCS keratinocytes, providing insights into how loss of CYLD function can affect biological processes in skin.
O04 Delineation of psoriasis from uninvolved skin using an artificial neural network as a means of targeting phototherapy to psoriatic lesions
Authors: Scott Joseph , Praeger Matthew , James Grant-Jacobs , Barnsley Josephine , Niranjan Mahesan , Eason Robert , Mills Ben , Healy Eugene 
Published: May 2024

Abstract

Introduction and aims
Narrowband ultraviolet B therapy (NB-UVB) is an effective treatment for psoriasis. Currently, NB-UVB is delivered in whole-body cabinets to all of the patient’s skin, thus increasing the risk of normal skin sunburning and the potential risk of skin cancer development. This limits the NB-UVB doses that can be used and requires patients attend sessions three times weekly for up to 10 weeks in order to complete treatment. An artificial intelligence (AI)-mediated, targeted NB-UVB approach would avoid these risks and allow use of higher NB-UVB doses for the areas of the skin affected by psoriasis. We sought to train an artificial neural network on digital images of psoriasis, and subsequently test this trained model in order to assess its accuracy at distinguishing psoriasis from unaffected skin.

Methods
Overall, 104 digital images of psoriasis from 14 patients with this skin condition, who attended the dermatology department at University Hospital Southampton NHS Foundation Trust, were used to train a conditional generative adversarial network– ‘pix2pix’. Areas of psoriasis were delineated within the images prior to the images being augmented by randomly cropping, rotating and altering colour intensity, thereby increasing the total number to 23 111 images to train and test the neural network.

Results
The trained neural network was able to identify psoriasis in unseen digital images with 96.73% accuracy at the pixel level across the dataset. The output of the neural network (i.e. a computer-generated image of psoriasis) acted as a signal source for a digital micromirror device (DMD) that was able to shape visible light and project this light over the original image of psoriasis, while avoiding the uninvolved skin.

Conclusions
It is possible to differentiate psoriasis from unaffected skin within digital images using an artificial neural network. We demonstrated that AI-mediated targeted phototherapy using a DMD is feasible.
O09 Microsatellite-unstable sporadic sebaceous and duodenal tumours are associated with loss of mismatch protein expression
Authors: Holt Georgie , Alfailakawi Waleed , Cook Sam , Ness Thomas , Coulthard Rowen , Jones Claire , Hayes Christine , Phelps Rachel , Kist Ralf , Jackson Mike , Santibanez-Koref Mauro , Burn John , Burt Alastair , Husain Akhtar , Lamb Chris , Gallon Richard , Rajan Neil 
Published: May 2024

Abstract

Introduction and aims
Lynch syndrome (LS) is a common heritable cancer predisposition syndrome. Tumours from patients with LS demonstrate microsatellite instability (MSI-high), a genetic signature detectable by low-cost DNA sequencing assays that is currently utilized in colorectal cancer screening. The utility of MSI assays for LS detection has not been extensively studied in unselected sebaceous skin tumours or duodenal tumours, where currently immunohistochemical (IHC) expression of mismatch repair (MMR) proteins is used. In addition, it has recently been suggested that cutaneous squamous cell carcinoma (cSCC) could be linked to LS; however MSI status has not been studied in patients with early-onset cSCC.

Methods
We investigated MSI status in archival formalin-fixed paraffin-embedded cases with ethical approval. We examined 110 sebaceous tumours (STs) (45 female patients, 65 male patients; median age 77 years), 50 duodenal tumours (DTs) (28 female patients, 22 male patients; median age 64 years), and 148 cSCCs selected from patients under 50 years of age (69 female patients, 79 male patients; median age 49 years). Immunohistochemistry (IHC) was performed on each MSI-high sample to determine the expression of MMR proteins MLH1, MSH2, MSH6 and PMS2. Samples that passed quality control were included in the analysis.

Results
A total of 42 of 106 STs (39.6%) were MSI-high. All STs had loss of at least one MMR protein. Overall, 10 of 41 (24%) DTs were MSI-high. All DTs had loss of at least one MMR protein. We found that 12 of 148 (8.1%) cSCCs were MSI-high and 5 of 12 SCCs exhibited loss of at least one MMR protein expression (41%).

Conclusions
MSI-high status correlated well with loss of MMR protein expression in STs and DTs samples. Future studies will help establish MSI status in tumours initially selected by MMR loss on IHC, informing its use as an adjunct to MMR IHC in LS detection in patients with STs and DTs.
O14 Monocytes and macrophages in the melanoma tumour microenvironment express Fc receptors which can be engaged with monoclonal antibodies of different isotypes
Authors: Adams Rebecca , Osborn Gabriel , Laddach Roman , Grandits Melanie , Chenoweth Alicia , Sanz-Moreno Victoria , Lacy Katie , Karagiannis Sophia 
Published: May 2024

Abstract

Introduction and aims
Melanoma is the deadliest form of skin cancer and has an increasing worldwide incidence. It is well established that the melanoma tumour microenvironment (TME) has a large immune cell infiltrate, including a prominent monocytic compartment. Monocytic cells within the TME support melanoma growth, survival and metastasis. Contrastingly, such cells can be stimulated to promote tumour killing when engaged with monoclonal antibodies (mAbs) against cancer antigens, owing to their expression of multiple Fc receptors (FcRs). However, little is known about the monocytic subsets present within melanoma, the FcRs they express and how engaging their FcRs may influence function.

Methods
Flow cytometry on peripheral monocytes isolated from patients with melanoma and healthy volunteers enabled the characterization of cell surface markers. Next, publicly available single-cell RNA-Seq datasets were analysed to explore the transcriptome of melanoma-associated macrophages and these phenotypes were recapitulated in vitro. Finally, monocytes and macrophages were cocultured with melanoma cells, with or without the addition of mAbs against a melanoma antigen, in order to assess the effect of FcR crosslinking on monocyte and macrophage phenotype and function.

Results
Circulating monocytes in patients with melanoma feature an inhibitory phenotype, and express multiple FcRs, including Fcε and Fcγ receptors, which could be targeted with mAbs of multiple isotypes. Secondly, monocytic cells within the melanoma TME express protumour genes, including metalloproteinases and angiogenic factors. However, FcR expression suggests that tumour-promoting subsets can be targeted with mAbs. Finally, in ex vivo cocultures with melanoma cells, the FcRs of both human monocytes and macrophages are able to engage antimelanoma mAbs of multiple isotypes, inducing differential phenotype change. This is demonstrated by the upregulation of activation markers and costimulatory cell surface markers.

Conclusions
Together, these data suggest that tumour-promoting monocytic cells within the melanoma TME can be engaged and harnessed using different isotypes of mAbs targeting melanoma.
O19 Improvement of a junctional epidermolysis bullosa mouse model
Authors: Jones Eleri , Garcha Priya , Aumailley Monique , O’Toole Edel , Rognoni Emanuel , Caley Matthew 
Published: May 2024

Abstract

Introduction and aims
Junctional epidermolysis bullosa (JEB) is the most severe of the epidermolysis bullosa (EB) disorders and is caused by mutations in genes encoding the skin basement membrane proteins laminin 332, type XVII collagen or integrin α6β4. Affected individuals experience blistering from birth leading to scarring, granulation tissue and susceptibility to infection. In JEB, generalized severe, there is 50% mortality in the first 2 years of life, often owing to failure to thrive or overwhelming infection. A number of animal models for JEB exist; however, because of severe blistering on the entire surface of the skin, these mice die soon after birth, reducing their experimental utility. In 2017, the first adult inducible knockout mouse model of JEB was published. In this mouse model, the disruption of the Lama3 gene is specifically induced in epidermal basal keratinocytes after birth. These mice remain viable, which allows us to study the longer-term consequences of the loss of laminin α3. However, the mice did not develop the severe phenotype observed in patients with JEB.

Methods
We have optimized this previously published methodology of a conditional knockout JEB mouse model (Lama3flx/flxK14CreERT) to now include more regular tamoxifen injections, which Results in a more controlled and severe phenotype of JEB.

Results
Widespread skin blistering develops after 12 weeks of tamoxifen treatment with an impaired skin barrier (transepidermal water loss) and a loss of epidermal lipids. Immunofluorescent staining was conducted to assess skin differentiation (K14, transglutaminase 1, loricrin), proliferation (Ki67), basement membrane proteins (laminin α3, laminin β3, laminin γ2, collagen VII) and immune markers (F4/80, Ly6-G/C, CD3, CD79).

Conclusions
These mice recapitulate all key features of human JEB with loss of laminin α3 at the basement membrane, epidermal thickening and visible blisters. This allows us to delve deeper into the functional and downstream pathways affected by the loss of laminin α3 and discover why the prognosis of patients with JEB is so severe.
P03 Anti-ageing effects of oestetrol on the skin: protection from cellular senescence and restoration of dermal architecture
Authors: Cooksey Leah , Fenercioglu Laden , Bernardes Tania , Fearn Alice , Gerard Céline , Dion Valérie , Hardman Matthew , Wilkinson Holly 
Published: May 2024

Abstract

Introduction and aims
Skin is a major endocrine organ, undergoing profound structural and functional alterations in response to hormonal changes. Postmenopause, the rapid decline in circulating 17β-oestradiol (E2) leads to skin ageing, while exogenous E2 administration restores skin function, promoting the regenerative capacity of aged skin. Nevertheless, the risk of off-target effects limits the clinical use of E2 for skin indications. Oestetrol (E4), a natural oestrogenic steroid currently undergoing clinical trials as a hormone replacement therapy, presents a favourable safety profile and could be a suitable alternative to E2. However, as no studies have investigated the effects E4 on skin ageing, the aim of this study was to evaluate the potential of E4 to overcome effects of skin ageing.

Methods
Micro-osmotic pumps were subcutaneously implanted into 14-month-old female mice, allowing sustained systemic delivery of E2 or E4 over 3 weeks. Reversal of skin ageing was assessed via noninvasive skin parameter testing, histological analysis and biomechanics. Aged human dermal fibroblasts were treated with E4 to measure effects on the extracellular matrix deposition (ECM) in vitro. Finally, human keratinocytes were treated with E4 prior to inducing senescence to assess potential protective effects against skin ageing. Readouts included senescence-associated beta galactosidase staining, immunofluorescence and quantitative polymerase chain reaction.

Results
Systemic administration of E4 reversed multiple characteristics of skin ageing. E4 treatment significantly increased skin hydration, stimulated collagen deposition and elevated elastic fibre synthesis. These structural changes improved the biomechanical function of skin, increasing breaking strength and elasticity. In vitro studies confirmed direct anti-ageing activity on multiple cell types. E4 substantially increased ECM deposition in aged human dermal fibroblasts and protected against senescence induction in human keratinocytes.

Conclusions
Collectively, these preclinical data demonstrate significant beneficial effects of E4 on aged skin, strongly supporting future clinical evaluation of E4 to alleviate the effects of skin ageing following menopause.
P05 The aryl hydrocarbon receptor pathway is dysfunctional in atopic dermatitis
Authors: Dawe Hannah , Tosi Isabella , Barker Jonathan , Smith Catherine , Di Meglio Paola 
Published: May 2024

Abstract

Introduction and aims
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by epidermal barrier disruption and type-2 immune dysregulation. While treatment has improved with targeted biologics, clinical response varies, highlighting the need for additional therapeutic strategies. The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor and environmental sensor, which maintains skin barrier integrity and homeostasis by reducing inflammation. Upon activation, AHR induces the expression of CYP1A1, a biomarker of pathway activation and critical for AHR ligand metabolism. Tapinarof, a topical AHR agonist, is approved for psoriasis and is effective in phase III clinical trials in AD, highlighting the importance of this pathway in inflammatory skin disease. Nevertheless, AHR expression is increased in AD skin, leading to the hypothesis that the pathway is dysregulated, thus not exerting its anti-inflammatory effect.

Methods
Here we begin to perform a global investigation of the AHR pathway in AD. We measured AHR and CYP1A1 mRNA expression by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) in healthy and AD skin, with spatial mRNA expression assessed by RNAscope. We also investigated AHR mRNA expression in interleukin (IL)-4 stimulated (100 ng mL−1) normal human epidermal keratinocytes (NHEKs) using qRT-PCR.

Results
We found that AHR mRNA is significantly upregulated in lesional AD (n = 6) compared with healthy skin (n = 9; P &lt; 0.05), whereas CYP1A1 mRNA expression shows a downward trend in lesional AD compared with healthy skin (P = 0.063). The AHR expression pattern differs in lesional AD (n = 7) compared with healthy skin (n = 8), with AHR localizing in the epidermal basal layer. Finally, IL-4 significantly upregulated AHR mRNA (n = 4 wells; P &lt; 0.001) expression in NHEKs compared with unstimulated cells.

Conclusions
Overall, our data have uncovered a dysfunctional AHR pathway in AD, with increased AHR expression, possibly driven by the AD inflammatory milieu, but reduced pathway activation. Further investigations are required to fully dissect this pathway and maximize its potential as a therapeutic target for AD.
P06 A novel drug combination produces a synergistic effect when inhibiting myofibroblast transformation in an in vitro model of hypertrophic scarring
Authors: Lapthorn Alice , Ilg Marcus , Dziewulski Peter , Cellek Selim 
Published: May 2024

Abstract

Introduction and aims
Hypertrophic scars affect 91% of patients with burns and yet, there is currently no effective treatment for their prevention. Myofibroblasts are key cells in the pathogenesis of hypertrophic scars, with the inhibition of myofibroblast transformation having been suggested as an effective therapeutic strategy. We have previously screened 1954 drugs approved by the Food and Drug Administration to identify any that could inhibit myofibroblast transformation. Using this data, the aim of this study was to investigate the individual and combined antimyofibroblast effect of two drug classes.

Methods
Primary human fibroblasts were isolated from the scars of patients with burns, with myofibroblast transformation induced using transforming growth factor (TGF)-β1. Concentration-response curves for drug-class A and drug-class B were constructed at a range of concentrations (0.03–300 µmol L–1), with the In-Cell enzyme-linked immunosorbent assay method used to measure α-smooth muscle actin (α-SMA) expression. Combination experiments were carried out through construction of a concentration-response curve for drug-class A (0.03–100 µmol L–1) in the presence of 3 µmol L–1 of drug-class B.

Results
Both drug classes showed a concentration-dependent antimyofibroblast effect with an IC50 range of 12.2–120.1 µmol L–1 for drug-class A and 1.4–16.7 µmol L–1 for drug-class B. When given in combination the two classes of drugs produced a synergistic effect at all concentrations up to 10 µmol L–1. For example, an 18% reduction α-SMA expression was expected with 1 µmol L–1 of drug-class A and 3 µmol L–1 of drug-class B; however, a 24% reduction in α-SMA expression was observed instead.

Conclusions
This is the first study to identify this synergistic effect between these two drug classes, and therefore this novel drug combination could be developed for the prevention of hypertrophic scar formation.
P12 Transcriptional and epigenetic changes drive increased cellular plasticity in keloid fibroblasts
Authors: Tekkela Stavroula , Hight-Warburton Willow , Shaw Tanya , Philpott Mike , O’Toole Edel , Rognoni Emanuel 
Published: May 2024

Abstract

Introduction and aims
Keloids are fibroproliferative scars that progressively grow beyond the original wound site. They occur as a result of genetic, epigenetic and environmental abnormalities. Fibroblasts are best characterized for their role in depositing and remodelling the extracellular matrix (ECM) and are believed to have a crucial role in keloid formation. However, the detailed molecular signatures that promote this pathogenic phenotype are still unknown.

Methods
To investigate the epigenetic and genetic changes in keloid fibroblasts, we performed assay for transposase-accessible chromatin (ATAC)-Seq and RNA-Seq on a patient-matched set of keloid and normal fibroblasts. ATAC-Seq is a high-throughput sequencing method that defines chromatin accessible regions in which the binding of sites of transcription factors that regulate gene expression can be mapped. Identified candidates are validated in keloid fibroblasts via quantitative polymerase chain reaction and their functional role is analysed in various in vitro assays.

Results
ATAC-Seq analysis revealed more accessibility in coding regions of keloid fibroblasts compared with normal fibroblasts. Pathways enriched in these regions include transcriptional regulation, embryonic development, cell differentiation, calcium-binding and collagen remodelling. Intriguingly, several developmental epigenetic modulators were identified that may alter the transcription and chromatin landscape of keloid fibroblasts to a more plastic state and have the capacity to influence cell differentiation choices, potentially promoting chondrogenic and adipogenic cell features. Currently, we are investigating the ability of keloid fibroblasts to be more readily differentiated into other cell types including induced pluripotent stem cells, and the contribution of those candidates on fibrotic features to the cell are also being explored.

Conclusions
Our analysis reveals that the chromatin of keloid fibroblasts is more accessible, especially in genomic regions associated with embryonic development and keloid fibroblasts show an increased plasticity in vitro. These findings may support the development of fibroblast-targeted therapies for keloid scars and possibly other connective tissue disorders.