bluuebunny/crossref_metadata_2025_split huggingface dataset:
Childhood and Life-Course Socioeconomic Position and Cognitive Function in the Adult Population of the Hispanic Community Health Study/Study of Latinos
Authors: Filigrana Paola , Moon Jee-Young , Gallo Linda C , Fernández-Rhodes Lindsay , Perreira Krista M , Daviglus Martha L , Thyagarajan Bharat , Garcia-Bedoya Olga L , Cai Jianwen , Lipton Richard B , Kaplan Robert C , Gonzalez Hector M , Isasi Carmen R 
Published: July 2023

Abstract
The Hispanic/Latino population experiences socioeconomic adversities across the lifespan and is at greater risk of cognitive impairment, yet little is known about the role of life-course socioeconomic position (SEP) in cognitive function in this population. Using baseline data (2008–2011) from adults (aged 45–74 years) of the Hispanic Community Health Study/Study of Latinos, we assessed the association between childhood SEP and socioeconomic mobility with cognitive function, and whether this association was mediated by midlife SEP. Childhood SEP was assessed using parental education. An index combining participants’ education and household income represented midlife SEP. Socioeconomic mobility was categorized as stable low, downward or upward mobility, and stable high-SEP. Cognitive function measures were modeled using survey linear regression with inverse-probability weighting, accounting for covariates. We used mediation analysis to estimate the indirect effect of childhood SEP on cognition through midlife SEP. High childhood SEP was associated with global cognition in adulthood (coefficient for parental education beyond high school vs. less than high school = 0.26, 95% confidence interval: 0.15, 0.37). This association was partially mediated through midlife SEP (indirect effect coefficient = 0.16, 95% confidence interval: 0.15, 0.18). Low SEP through the life course was associated with the lowest cognitive function. This study provides evidence that life-course SEP influences cognitive performance in adulthood.
O05 Functional roles of lncRNA in cutaneous squamous cell carcinoma disease progression
Authors: Bone Max , Harwood Catherine , Leigh Irene , Bailey Peter , Inman Gareth 
Published: July 2023

Abstract
Cutaneous squamous cell carcinoma (cSCC) is one the most prevalent malignancies with metastatic potential within the UK and is of significant public concern due to its increasing annual incidence. The progression of cSCC has been historically characterized into several key stages, but the mechanistic properties as to how the disease advances remains poorly defined. In recent years, long noncoding RNA (lncRNA), a widely uncharacterized and poorly understood family of RNA molecules over 200 nucleotides in length, have become of increasing interest in regulating epigenetics and various other cellular processes. These molecules have also been found to be widely associated with cancer pathways as both inhibitors and drivers of disease, and have been identified in all cancers as potential molecules of interest, including cSCC. Although several lncRNA have been recognized as functionally relevant in cSCC, they represent a small proportion of the estimated 300 000 lncRNA transcripts of which most have unknown functions within both skin and cSCC specifically. By performing RNAseq on 110 patient samples, including normal sun-exposed skin (n = 26), actinic keratosis (n = 14), primary cSCC (n = 66) and metastases (n = 4) and utilizing various bioinformatic analysis pipelines, key lncRNA of interest were identified in cSCC. These lncRNA of interest were then explored in vitro in a panel of patient-derived cSCC cell lines to find key lncRNA of functional relevance. These functionally active lncRNA were subjected to further in silico analysis to determine splice isoforms and transcripts responsible for disease progression, and potential mechanisms of regulating cellular epigenetics.
O07 Filaggrin insufficiency renders keratinocyte-derived small extracellular vesicles capable of affecting CD1a-mediated T-cell responses and promoting allergic inflammation
Authors: Kobiela Adrian , Hewelt-Belka Weronika , Frąckowiak Joanna E , Kordulewska Natalia , Hovhannisyan Lilit , Bogucka Aleksandra , Etherington Rachel  , Piróg Artur , Dapic Irena  , Gabrielsson Susanne , Brown Sara , Ogg Graham S   , Gutowska-Owsiak Danuta 
Published: July 2023

Abstract
The association between FLG loss-of-function mutations and the development of atopic dermatitis (AD) indicates that filaggrin is critical to skin barrier function; the mutations are also linked to additional allergic manifestations, but it is unknown how the skin may influence inflammation in distant tissues. Here we investigated the impact of filaggrin insufficiency on keratinocyte-derived exosome-enriched small extracellular vesicles (sEVs) and found that those secreted by filaggrin-knockdown cells are extensively remodelled as a consequence of the abnormal keratinocyte differentiation process. The altered sEV lipidome changes the sEV capacity to promote T-cell responses by direct effect on self-lipid neoantigen generation, modulating the content of permissive (stimulatory) and nonpermissive (inhibitory) CD1a ligands released from the sEV membranes by phospholipase A2. Presentation of those lipids to T cells results in the redirection from a type 1 to a type 2 response. We found that the aberrant sEV lipid composition mirrors a generalized remodelling of cellular lipids resulting from dysregulated expression of multiple enzymes of lipid metabolic pathways, observed both in filaggrin knockdown keratinocytes in vitro and in the skin of patients with AD. Hence, filaggrin insufficiency background renders keratinocytes capable of supplying CD1a ligands which are capable of impeding both homeostasis-promoting and protective antimicrobial responses, enhancing type 2 T-cell activation to perpetuate inflammation in AD skin. sEV transfer by systemic circulation may promote similar effects in distant tissues.
O14 Junctional epidermolysis bullosa: repairing the epidermal lipid barrier
Authors: Jones Eleri , Marsh Supatra , O'Shaughnessy Ryan , Aumailley Monique , McGrath John  , O’Toole Edel  , Caley Matthew 
Published: July 2023

Abstract
Junctional epidermolysis bullosa (JEB) caused by loss of function mutations in basement membrane genes, including that encoding laminin-332, is among the most severe forms of epidermolysis bullosa. Affected individuals suffer from blistering from birth, leading to scarring, granulation tissue and susceptibility to infection. In generalized severe JEB, there is a 50% mortality rate in the first 2 years of life, often due to failure to thrive or overwhelming infection. Using cells with knockdown of laminin-α3 we generated a RNAseq data set and have identified a new pathway altered in JEB, the regulation of cholesterol biosynthesis. It has been shown that loss of laminin-332 results in reduced epidermal lipid barrier specifically due to loss of cholesterol transport within keratinocytes. Cholesterol biosynthesis and cholesterol trafficking are fundamental processes in a wide range of cells and tissues, especially in the formation of the normal epidermal lipid barrier. A cholesterol transport screening assay has been developed and used to analyse the ability of 10 specifically selected compounds to restore cholesterol transport in our in vitro model of JEB. Results of this small-scale screen showed that nine of 10 of the selected drugs were able to restore cholesterol transport up to 50% of control. Six drugs were selected for further analysis within our three-dimensional (3D) skin-equivalent model of JEB. Organotypic skin equivalents of normal skin (siControl) and JEB skin (siLaminin-α3) were treated daily for 7 days with either dimethyl sulfoxide or 1 µmol L–1 of selected drugs. In 3D, drugs 1, 3, 5 and 6 displayed an increase in stratum corneum lipids shown by Nile red staining. Drugs 4 and 7 did not increase stratum corneum lipids above levels seen in the untreated JEB organotypics. Further analysis of skin differentiation by keratin, involucrin and tranglutaminase staining were assessed to narrow drug selection for further testing. These data also suggest that JEB is not just a disorder of epidermal–dermal adhesion, but a disorder of aberrant cholesterol trafficking causing a markedly impaired skin barrier. These findings suggests that drugs to target cholesterol transport within keratinocytes could be a beneficial treatment for JEB, helping to improve the crucial epidermal lipid barrier and prevent water loss and infection at the early stages of life.
O18 Molecular crosstalk between PLCγ1 and STAT3 in cutaneous T-cell lymphoma
Authors: Campbell Victoria , Flanagan Charlotte , Tosi Isabella  , Jones Christine , Whittaker Sean , John Susan , Mitchell Tracey 
Published: July 2023

Abstract
Cutaneous T-cell lymphoma (CTCL) is a non-Hodgkin lymphoma of skin-homing T lymphocytes that is difficult to diagnose and treat due to a high degree of clinical and genomic heterogeneity. Mutations in PLCG1/PLCγ1 occur in 10% of CTCL tumours. Meanwhile, signal transducer and activator of transcription 3 (STAT3) – a common driver of lymphomagenesis – is frequently activated (pY705-STAT3) in CTCL in the absence of mutation. Activation of STAT3 may also be mediated by serine phosphorylation (pS727-STAT3), and there is a lack of data on the role of this modification in CTCL. We aimed to identify if gain-of-function PLCG1 mutations in CTCL activate STAT3, either directly or indirectly, via downstream mitogen-activated protein kinase (MAPK) signalling. Flow cytometry was used to study STAT3 phosphorylation and subcellular localization in ex vivo CD4+-enriched tumour cell populations from patients with CTCL and healthy donors. STAT3 phosphorylation was also studied in the J.gamma1 T-cell line (PLCγ1-null) and primary ex vivo CD4+ cells upon lentiviral overexpression of wild-type (WT) and mutant (p.R48W, p.S345F) PLCγ1 constructs. To investigate the role of MAPK signalling, the CTCL-derived Hut78 and SeAx cell lines were treated with MAPK inhibitors (trametinib), and STAT3 phosphorylation assessed by Western blotting. Significantly increased basal levels of pS727-STAT3 (P &lt; 0.05) and pY705-STAT3 (P &lt; 0.001) were observed in CD4+-enriched populations from patients with Sézary syndrome compared with healthy donors. Mitochondrial translocation of STAT3 was also increased in CD4+-enriched populations from patients with Sézary syndrome (P &lt; 0.05). In transduced J.gamma1 cells, p.S345F-PLCγ1 overexpression activated pS727-STAT3 (P &lt; 0.05), while overexpression of both WT and mutant PLCγ1 constructs activated pS727-STAT3 in primary ex vivo CD4+ cells (P &lt; 0.05 and P &lt; 0.01, respectively). Inhibition of MAPK kinase (MEK)–extracellular signal regulated kinase (ERK) signalling did not significantly modulate STAT3 phosphorylation or subcellular localization in CTCL cell lines, suggesting that pS727-STAT3 phosphorylation is not mediated via ERK in CTCL. This work extends our understanding of PLCγ1 signalling and identifies pS727-STAT3 as a potential target of interest for further study in CTCL.
PostersP01 Tissue slice culture as a patient-derived explant model to investigate immunity and immunotherapy in cutaneous squamous cell carcinoma
Authors: Coltart George , Lai Chester , Al-Shamkhani Aymen , Healy Eugene 
Published: July 2023

Abstract
Cutaneous squamous cell carcinoma (cSCC) is one of the commonest cancers worldwide with the capacity to metastasize. Immune evasion by the tumour prevents its elimination and allows it to progress, including enabling the cancer to spread to local lymph nodes and distant organs. Targeting this dysfunctional immunity is a key strategy for the treatment of cSCC and anti-PD-1 checkpoint inhibition is now the first-line therapy for advanced and metastatic cSCC. However, complete responses to anti-PD-1 are seen in &lt; 15% of patients and therefore it is vital that other strategies for enhancing anticancer immunity in cSCC are explored. Given the expense of clinical trials, it is essential to have good preclinical data to target cancer drug development appropriately, and this can be challenging in the investigation of cancer immunotherapy, as these treatments rely upon an intact human immune–tumour interface. We have developed a tumour slice culture (TSC) system to investigate the mechanism and efficacy of immune-modulating anticancer therapy in cSCC. This patient-derived explant model involves taking tissue slices from freshly excised tumours and culturing the slices under different experimental conditions, while maintaining the tumour microarchitecture of each slice. We cultured &gt; 50 tumours using the TSC platform, including 35 cSCCs. Immunofluorescent confocal microscopy of anti-CD4 demonstrates penetration of antibody throughout the tissue slice. The mean (SD) viability of lymphocytes (T and B cells) determined by flow cytometry after 72 h of culture was 90.3% (7.89%; n = 31). Treatment of slices with OKT3 anti-CD3 antibody for 72 h induces a dose–response increase in T-cell Ki67 expression (2.87% unstimulated vs. 13% OKT3 100 ng mL–1; P &lt; 0.001) and CD8+ T-cell granzyme B (47.6% unstimulated vs. 63.7% OKT3 ng mL–1; P = 0.064), as detected by flow cytometry (n = 7). Enzyme-linked immunosorbent assay of the culture supernatant showed that interferon-γ concentrations increased from &lt;9.6 pg mL–1 (unstimulated) to 131.1 pg mL–1 [OKT3 1000 ng mL–1 (n = 6, P &lt; 0.05]. To our knowledge the TSC system has not previously been used in cSCC and represents a promising technique to investigate new potential immunotherapies in this cancer.
P10 Inducible 14-3-3 sigma/stratifin ablation cooperates with rasHa activation to accelerate papillomatogenesis and induce malignant conversion in transgenic mouse skin carcinogenesis
Authors: Hardman-Carter Rebecca , Lefkati Revvekka , McMenemy Carol  , Greenhalgh David 
Published: July 2023

Abstract
In carcinogenesis, the 14-3-3σ isoform (stratifin) is implicated by being downregulated in a mode of classic tumour suppression gene (TSG) loss and also via elevated oncogenic roles; with both scenarios appearing to be hallmarks of 14-3-3σ deregulation, depending on tumour stage or context. Mouse skin models highlight tumour-suppressive roles, as the Er/Er+/– repeated epilation stain possesses germline mutations that result in hyperplasia, failed follicular differentiation and susceptibility to squamous cell carcinoma (SCC). In transgenic models, 14-3-3σ knockout supports early roles, resulting in rapid papilloma formation in classic two-stage DMBA/TPA chemical carcinogenesis; however, our previous studies in this model demonstrated targeted overexpression of 14-3-3σ led to SCC of a follicular origin in collaboration with Fos and well-differentiated SCC in collaboration with rasHa. Therefore, to study mechanisms driving/inhibiting skin carcinogenesis, stage-specific expression of endogenous 14-3-3σ (stratifin) was analysed in early skin carcinogenesis driven by activated rasHa expression (HK1.ras) and subsequently via conditional RU486-inducible 14-3-3σ knockout (K14.creP-Δ14-3-3σ). Consistent with 14-3-3σ roles previously observed in epidermal differentiation, HK1.ras hyperplasia and early papillomas displayed elevated 14-3-3σ expression in suprabasal keratinocytes and in occasional K1-positive basal keratinocytes as they committed to differentiate. Older HK1.ras mice exhibited increasing 14-3-3σ positivity in papilloma basal layers, suggesting attempts to protect basal-layer p53 TSG function, given that 14-3-3σ acts as a chaperone protein to remove/relocate MDM2 for degradation, thus maintaining p53 levels. Indeed, such papillomas expressed activated p-MDM2166 confined to suprabasal layers, resulting in strong basal layer p53 expression and thus HK1.ras papillomas lacked malignant conversion. In mice where 14-3-3σ functions were ablated via an RU486-inducible Cre/loxP system expressed from a keratin 14 (K14) promoter (K14.creP-Δ14-3-3σ), treated HK1.ras-K14.Δ14-3-3σ mice exhibited a rapid papillomatogenesis and produced larger papillomas. These papillomas also exhibited rapid conversion to malignancy, associated with increased activated p-MDM2166 expression in basal layers and resultant p53 loss. These observations suggest that 14-3-3σ plays tumour suppressive roles involving p53 protection, where failure to regulate MDM2 activity led to p53 loss and susceptibility to malignant conversion.
P11 Abstract withdrawn.
P12 Tetrapeptide matrikines synchronize circadian rhythms and promote proliferation in human skin cells
Authors: El-Houni Zeyad , Eckersley Alexander , Bell Mike , Bradley Eleanor , Newton Victoria , Sherratt Michael  , Meng Qing Jun 
Published: July 2023

Abstract
Circadian rhythms are intrinsically generated 24 h rhythms that play a vital role in enabling the skin to adapt temporally to various external stimuli. These include ultraviolet radiation, physical trauma and water loss. At the molecular level, circadian clocks consist of various clock genes and proteins, including Per2/Cry1 and Bmal1/Clock, forming autoregulatory feedback loops. Although these rhythms are known to be impaired in ageing skin, there are limited therapeutic options to restore skin cell circadian function. As small peptides with homology to amino acid sequences found in extracellular matrix proteins (matrikines) can beneficially affect clinical and histological markers of skin ageing, in this study we characterized the ability of two novel tetrapeptides to influence the cellular circadian clocks of skin cells. HaCat keratinocytes transduced with a human Per2::luc promoter-driven reporter were used to monitor the effects of two synthetically generated peptides, GPKG and LSVD, on the cells’ intrinsic circadian clock. Following administration of these peptides at various concentrations (4–16 ppm), the oscillatory activity of the cells was amplified up to twofold vs. the vehicle control. LSVD was found to induce Per2::luc promoter activity significantly (P &lt; 0.05). In addition to enhancing circadian rhythmicity, both peptides were assessed for their proliferative effect (by Incucyte imaging and MTT cell proliferation assays). At the higher concentrations tested, both GPKG and LSVD significantly increased proliferation in HaCat keratinocytes (P &lt; 0.05). In conclusion, these peptides have been found to be highly effective in synchronizing the circadian clock in skin cells, leading to changes in their behaviour. This implies that these new peptides may have beneficial effects on skin cells, in part, by improving their circadian function. Given the findings of clock disruption in a number of diseases, as well as with age, identifying compounds that can reset the clock in skin cells is an important approach to developing treatments that can improve the functioning of cutaneous physiological processes that are under circadian control.
Funding sources: this study was funded by the No. 7 Beauty Company.
P17 A signature of IL-36 activation in systemic lupus erythematosus skin
Authors: Sumner Emma J , McCluskey Daniel , Mahil Satveer K   , Lewis Myles , Capon Francesca  
Published: July 2023

Abstract
Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease with heterogeneous presentation. Joint and kidney disease represent the largest cause of morbidity, and visually distinctive skin complications are often present. While it has long been established that organ damage is caused by autoantibodies forming large immune complexes in the affected tissues, a detailed understanding of disease processes is still lacking. Here, we propose that interleukin (IL)-36 cytokines, which drive cutaneous and systemic symptoms in psoriasis, are also implicated in the pathogenesis of SLE. Given that the IL-36 receptor is prominently expressed in keratinocytes, we investigated this hypothesis in skin. We first carried out a systematic review of SLE single-cell RNA sequencing studies, identifying two data sets (∼50 000 cells in total) generated from patient skin biopsies. We processed both data sets with the same standardized pipeline, using the Seurat package. This identified 10 distinct cell clusters, replicating the cell types described in the original studies and identifying the main cell populations observed in human skin. Subsequent gene expression analysis confirmed that, in both data sets, the expression of IL36G and IL1RL2 (the genes encoding IL-36γ and its receptor) was highest in keratinocytes. Interestingly, subclustering showed that this signal was driven by the expression of IL36G in supraspinous keratinocytes and of IL1RL2 in proliferating keratinocytes. In the first data set, the expression of IL36G and IL1RL2 was readily detectable in lesional keratinocytes but virtually nonexistent in their nonlesional counterparts. A comparison of lesional and nonlesional keratinocytes also identified 297 differentially expressed genes (DEGs; log2 fold change &gt; 1.2, false discovery rate &lt; 0.05) in the first study and 234 in the second. To further explore these findings, we compared the above DEG with a set of 182 genes (previously defined by our group) that are upregulated in IL-36-stimulated keratinocytes. This demonstrated a significant over-representation of the IL-36 signature genes among the DEGs observed in lesional SLE keratinocytes (P &lt; 0.005 in both data sets). These observations indicate that IL-36 is likely to contribute to skin inflammation in SLE, warranting further studies of this cytokine in organs affected by the disease.