tbuckley/cabot-search huggingface dataset:
P112 An exclusive established cell model of patient-derived psoriatic keratinocyte provides a valuable <i>in vitro</i> tool for mechanistic studies and therapeutic testing
Journal: British Journal of Dermatology (2024)

Abstract Chronic inflammatory skin conditions, including psoriasis, result in a wide range of distressing symptoms for affected individuals. There is a pressing need for epidermal models which can be readily utilized, enhancing our comprehension of these disorders. We report, the establishment of a novel psoriatic keratinocyte cell line that embodies essential features of psoriasis. Psoriatic keratinocytes (PSA cells), isolated from epidermal hypertrophic plaques of a 50-year-old male with psoriasis vulgaris, and a control keratinocyte (KT2) cell line, derived from an aged-matched healthy human skin sample, were immortalized with SV40-large T antigen. The transcriptomic and biological analysis of these established cellular models revealed significant phenotypic distinctions between healthy and psoriatic keratinocytes. Our findings from RNA-seq data analysis (validation by qRT-PCR) demonstrate that PSA cells from psoriatic individuals display a heightened expression profile of inflammatory cytokines and peptides (including TNF-α, IL-1β, IL-23, IL-8, LL-37, and serpin B1) compared with KT2 cells from healthy individuals, mirroring the clinical characteristics observed in psoriasis patients. Stimulation of the cells for 24 h with phorbol myristate acetate (PMA) which induces reactive oxygen species (ROS) production and protein kinase C (PKC) activation, ultimately leading to an inflammatory response further increased most of cytokine’s expression in PSA cells when compared with non-treated KT2 cells. This confirms that the psoriatic cell model is capable of launching additional inflammatory responses upon exposure to stress factors, in addition to their already innate inflammatory nature. Additional comparative analysis of PSA cells and KT2 cells in conjunction with NTERT cells, a healthy keratinocyte line conventionally used in cutaneous research, found that the PSA cells exhibited reduced expression of anti-inflammatory glucocorticoid receptor, comparable to what detected in primary keratinocytes isolated from psoriatic lesional and non-lesional skin. Further transcriptomic and morphological examination of these newly established cell models revealed enlarged nuclear and cell body size and a faster growth rate of PSA cells in comparison with either the healthy NTERT or KT2 cell lines, all trademark affiliations of psoriasis. These findings suggest that immortalized psoriatic PSA cells, the first of their kind, faithfully profile the psoriatic phenotype in vitro and may provide a novel tool for interrogating the disease's pathogenic mechanisms.
FC28 Age-related changes in psoriasis severity and morphology
Journal: British Journal of Dermatology (2024)

Abstract The natural progression of psoriasis with age remains under-researched despite the ageing population and the increasing prevalence of this chronic condition. In this study, we examined how psoriasis changes over time, particularly in older adults. An online survey was conducted among members of the UK Psoriasis Association aged 65 years and older who had any symptoms of psoriasis before the age of 40. Participants rated their psoriasis severity on a scale from 0 (none) to 10 (severe) and provided details about their psoriasis treatments, lifestyle habits, medications, and associated medical conditions at different life stages in 20-year intervals from birth up to 60 years, and 5-year intervals from the age of 61 and beyond. Open-ended questions were included to explore changes in psoriasis morphology and distribution over time. Generalized Estimating Equations model analysed changes in disease severity, using ages 21–40 as reference. Predictors of high disease severity (≥5) were identified and validated via logistic regression and backward stepwise selection. Thematic qualitative analysis was conducted by a dermatologist and a behavioural sciences scholar. Of 615 eligible members of the Psoriasis Association, 194 completed the survey (response rate 31.5%). The final analysis included 180 participants who met the inclusion criteria (56.1% male, 43.9% female, median age of 73 years). Results showed a significant decrease in self-rated psoriasis severity with age, reaching the lowest severity in individuals over 80 years old (mean severity, 2.8), with a highly significant reduction across all age categories over 60 (P &amp;lt; 0.001). Less severe disease in individuals over 65 was associated with longer disease duration (P = 0.012) and statin use (P = 0.029), while more severe disease correlated with itch (P = 0.003). Qualitative analysis identified three main themes: reduced severity (less redness, smaller and thinner plaques, decreased scaliness and itch), changes in distribution (from widespread to localized, with decreased scalp involvement and increased truncal involvement), and retirement as a factor in disease improvement. Our findings indicate that psoriasis severity reduces with age, particularly after 60, and that psoriasis morphology and distribution change over time, from large to smaller and thinner plaques, and from widespread to localized disease. The reduction in severity was associated with longer disease duration and might be attributed to the waning of the immune system and immunosenescence. Immunosenescence primarily affects T cells, which are integral to the pathogenesis of psoriasis, with evidence suggesting that inflammageing involves a shift towards a Th2 immune profile, which could be disease-ameliorating in psoriasis.
P56 Validation and comparison of the early arthritis for psoriatic patients questionnaire, the psoriasis epidemiology screening tool and the psoriatic arthritis uncluttered screening evaluation (PURE-4) in a Danish population
Journal: British Journal of Dermatology (2024)

Abstract Up to 30% of patients with psoriasis (PsO) develop psoriatic arthritis (PsA). National guidelines recommend screening patients with PsO for PsA; however, there is no consensus on the best-suited method. This study aims to validate and compare three international PsA screening questionnaires (PEST, EARP and PURE-4) in a Danish setting. The English questionnaires were translated and culturally adapted to the Danish language. Adult patients with PsO and without established PsA were recruited from a dermatology department at a university hospital, a private dermatology clinic and the Danish Psoriasis Association. The patients answered the three screening questionnaires in a random order, and all patients were subsequently seen by a rheumatologist who ruled out or verified the diagnosis of PsA according to the CASPAR criteria. The performance of each questionnaire was assessed by determining the sensitivity and specificity at different cut-off values and analysing the receiver operator characteristic (ROC) curves. A total of 114 patients participated in the study. At a cut-off value of ≥3, the PEST questionnaire had a sensitivity of 66.7% and specificity of 67.6%, while the numbers for EARP were 88.9 and 39.0%, respectively. The PURE-4 scale had a sensitivity of 100% and specificity of 26.7% at a cut-off value of ≥ 1. The area under the curve (AUC) for PEST was 0.71 [95% confidence interval (CI): 0.51–0.92]. For EARP, the AUC was 0.75 (95% CI: 0.58–0.91), while PURE-4 had an AUC of 0.67 (95% CI: 0.53–0.82). Optimal cut-off values were determined to be ≥3, ≥6 and ≥2 for PEST, EARP and the PURE-4 scale, respectively. When looking at the questionnaires’ ability to find patients with either rheumatologically verified PsA or probable PsA, meaning likely PsA but not complete fulfilment of the CASPAR criteria, all questionnaires maintained almost the same sensitives and specificities. The questionnaires were successfully cross-culturally validated and had an acceptable performance in line with other translated versions. However, all three questionnaires performed worse than in the original validation studies. The EARP questionnaire had the best discriminatory ability; however, a higher cut-off point for referral should be considered.
FC13 Comparative analysis of the oral microbiota in patients with psoriasis, their cohabiting partners, and healthy controls
Journal: British Journal of Dermatology (2024)

Abstract Accumulating evidence supports an association between altered oral microbiota and inflammatory diseases. However, the existing literature on the role of the oral microbiota in patients with psoriasis is limited. Previous studies demonstrated conflicting results and have never been based on shotgun metagenomic sequencing. Several differences have been found in taxa abundance between patients and controls, and culture-based studies all found increased levels of Candida species in patients with psoriasis. The objective was to investigate the differences in the composition and the functional potential of the oral microbiota in patients with psoriasis compared with their partners and healthy controls. A case–control study was conducted with 123 participants divided into three groups; patients with psoriasis not receiving systemic anti-psoriatic treatment (n = 52), cohabiting partners of the patients (n = 21), and age-, sex- and body mass index-matched healthy controls (n = 50). One saliva sample was collected from each participant and analysed by shotgun metagenomic sequencing. Statistical analyses included a comparison of α- and β-diversity, as well as an abundance of the different taxa and Kyoto Encyclopedia of Genes and Genomes (KEGG) modules. A total of 115 samples were included in the statistical analysis, divided into patients with psoriasis (n = 47), partners (n = 21) and healthy controls (n = 47). No significant differences in baseline characteristics were found between patients with psoriasis and their partners. Smoking status (P &amp;lt; 0.0001) and physical activity levels (P = 0.0071) differed between the psoriasis and control groups. We observed a significant difference in α-diversity in the oral microbiome between patients with psoriasis and controls (P = 0.014), showing lower diversity in patients with psoriasis. β-Diversity analysis showed shifts in composition due to smoking (P = 0.014), but no other investigated variable had a significant association with the β-diversity, although the psoriasis group tended to cluster together apart from the two control groups. Abundance analysis was performed using both Mann–Whitney U-test and linear regression, with the former method showing one species (Leptotrichia sp018128225, FDR = 0.079) and one family (Leptotrichiaceae, FDR = 0.059) significantly different between patients and controls, whereas the latter method identified two taxa significantly different between psoriasis and control groups, as well as 21 taxa and 1 KEGG module significantly different between the psoriasis group and their partners at an FDR-adjusted P-value cut-off of 0.1. No differences were found in the abundances of Candida species. A comparison of the microbiomes of individuals with psoriasis and controls indicates that psoriasis is associated with changes in oral microbiota diversity, composition and function.
P53 Guselkumab binding to CD64+ IL-23–producing myeloid cells enhances potency for neutralizing IL-23 signalling
Journal: British Journal of Dermatology (2024)

Abstract IL-23 is implicated in the pathogenesis of psoriasis (PsO), and myeloid cells expressing FcγRI (CD64) are the primary source of IL-23 in lesional PsO skin tissue. The incidence and prevalence of psoriatic arthritis increases with PsO severity, and joint disease activity is positively correlated with frequency of peripheral CD64+ monocytes. We evaluated: CD64 and IL-23 expression in PsO patient skin biopsies; binding of anti-IL-23p19 subunit IgG1 monoclonal antibodies (mAbs) guselkumab [(GUS) fully human, native Fc-region] and risankizumab [(RZB) humanized, mutated Fc-region] to CD64; and functional consequences of CD64 binding by anti-IL-23p19 subunit mAbs in in vitro assays. CD64, IL-23p19 and IL-23p40 subunits mRNA transcripts were analysed from bulk and single-cell RNAseq datasets. Binding of mAbs to IFNγ-primed human monocytes and IL-23–secreting inflammatory monocytes, and capture of endogenously secreted IL-23, were evaluated by flow cytometry. Internalization of IL-23 and GUS/RZB within CD64+ macrophages was assessed with live-cell confocal imaging. GUS and RZB potency for inhibiting IL-23 was determined in a co-culture of THP-1 cells (CD64+ monocyte cell line activated to produce IL-23) and an IL-23 reporter cell line (measuring biologically active IL-23). IL-23p19 mRNA transcript expression in the co-culture was measured by qPCR. Expression of CD64, IL-23p19, and IL-23p40 mRNA transcripts was increased in lesional vs. non-lesional PsO skin, as were the abundance of myeloid cell types co-expressing CD64/IL-23p19 mRNA transcripts. In in vitro assays, GUS (not RZB) demonstrated Fc-mediated binding to CD64 on IFNγ-primed monocytes. Moreover, CD64-bound GUS simultaneously captured IL-23 secreted from the same cells. GUS (not RZB) bound to the CD64+ macrophages and mediated IL-23 internalization to low pH intracellular compartments. GUS and RZB equipotently inhibited signalling by IL-23 present in THP-1-conditioned medium. However, in a co-culture of IL-23–producing THP-1 cells with an IL-23–responsive reporter cell line, GUS demonstrated enhanced potency vs. RZB for inhibiting IL-23 signalling. GUS did not alter IL-23p19 mRNA transcript expression in the co-culture. These findings were consistent with previous observations of CD64+ myeloid cells as a key source of IL-23 in lesional PsO skin tissue. GUS binding to CD64 on IL-23-producing cells likely contributed to the enhanced potency of GUS vs. RZB in inhibiting IL-23 signalling in the co-culture assay. These in-vitro data support a hypothesis for optimal localization of GUS in inflamed tissues, where CD64+ IL-23-producing myeloid cells are increased and in proximity to IL-23-responsive lymphoid cells, enhancing GUS neutralization of IL-23 at its source of production.
Concomitant Administration of Dantrolene is Sufficient to Protect Against Doxorubicin-Induced Cardiomyopathy
Authors: Yoshihide Nakamura, Takeshi Yamamoto, Shigeki Kobayashi, Takeshi Suetomi, Hitoshi Uchinoumi, Tetsuro Oda, Motoaki Sano, Masafumi Yano
Journal: JACC CardioOncology (2024)

Doxorubicin (DOX), a commonly used anticancer agent, can result in cardiac dysfunction, presenting a significant clinical challenge. DOX has been shown to induces Ca2+ leakage via the ryanodine receptor 2 (RYR2) of the sarcoplasmic reticulum, increasing Ca2+ levels in the cytoplasm. This study investigated whether stabilizing RYR2 could suppress DOX-induced cardiomyopathy (DIC) and identified the optimal duration of dantrolene treatment as a pharmacological method. We investigated the effects of RYR2 stabilization on DOX cardiotoxicity using in vivo and in vitro experiments. DOX administration caused calmodulin dissociation, marked Ca2+ leakage from RYR2, and increased oxidative stress in isolated cardiomyocytes. Stabilizing the RYR2 tetramer-either pharmacologically with dantrolene or genetically via RYR2 V3599K mutation, which enhances calmodulin binding affinity-suppressed these effects. In DIC mice models, DOX impaired cardiac function, increased fibrosis and TUNEL-positive cells, reduced GRP78, and elevated lipid peroxide levels, leading to endoplasmic reticulum stress and ferroptosis. Both continuous dantrolene treatment and RYR2 V3599K mutation improved cardiac function. Interestingly, dantrolene administration provided myocardial protection even when terminated 7 days after DOX. Short-term concomitant use of dantrolene offers a promising and clinically feasible strategy to prevent DIC. Given dantrolene's established clinical safety as a treatment for malignant hyperthermia, these findings suggest potential for repositioning dantrolene in DIC prevention.
P07 Shared pathophysiological pathways indicating treatment success of pityriasis rubra pilaris using anti-psoriatic anti-interleukin-17 therapy: a case report
Journal: British Journal of Dermatology (2024)

Abstract Pityriasis rubra pilaris (PRP) is a rare and severe inflammatory skin disorder characterized by reddish-orange scaly patches with prominent follicular involvement. PRP is classified into six distinct clinical subtypes, one of which is genetic. This genetic subtype arises from a gain-of-function mutation of the CARD14 gene activating nuclear factor-kappa B (NFkB) typically located within the psoriasis susceptibility locus 2 (PSORS2) activating the IL13/IL17A cytokine axis.1 Similar to psoriasis, activation of the Th17/IL-23 inflammatory pathway presents diverse therapeutic opportunities beyond conventional therapies.2,3 Traditional therapies, such as high-dose vitamin A or methotrexate, are often associated with a number of adverse effects and limited tolerability.4 We present a case of a 35-year-old female with an unusual presentation of PRP-mimicking guttate psoriasis who showed remarkable improvement with an anti-psoriatic, anti-interleukin (IL)-17A antibody(Cosentyx®, Secukinumab, Novartis). Complete remission was achieved within 8 weeks of treatment and continued to display sustained efficacy and good tolerability. This case highlights the importance of considering PRP in the differential diagnosis of psoriasis-like lesions and the potential efficacy of targeted biologic therapies in its management.
P80 Inhibiting HuR/ELAVL1 activity as a novel anti-psoriatic pharmacological strategy: in vitro evidence
Journal: British Journal of Dermatology (2024)

Abstract HuR is among the most widely studied RNA binding proteins that post-transcriptionally regulate thousands of coding and non-coding RNAs bearing AU-rich elements (AREs). It exerts both pre-mRNA processing and maturation functions and regulation of mRNA stability and translation. Interestingly, most of the chemokines and cytokines contain AREs, suggesting that HuR-dependent post-transcriptional control is crucial for the inflammatory response. HuR is ubiquitously expressed, and the outcome of its activation is context-dependent, and it is frequently deregulated in many types of cancers and immune-related diseases. Psoriasis is characterized by an intricate feed-forward interaction between the immune system and the skin through inflammatory cytokines that were identified as post-transcriptionally regulated by HuR. Indeed, HuR is often hijacked in diseases as in psoriatic keratinocytes. In Th17, HuR is responsible for the expression of specific key molecules for psoriasis pathogenesis. It has been shown that 15,16 dihydrotanshinone (DHTS) specifically inhibits the HuR-RNA complex formation. Recently, we have developed a new class of small molecules (named tanshinone mimics, TMs), bearing various substitutions, that modulate the interaction between HuR and its target RNAs in mammalian cells in vitro, more potently than the original DHTS, while displaying more druggable properties. We investigated the role of HuR in psoriasis both at the keratinocytes and at the T-cell levels in patients derived primary cells. HuR is increased in keratinocytes isolated from lesional psoriatic skin compared with healthy subject human keratinocytes, at the protein level. Silencing HuR expression in keratinocytes induced an increase of cells in the G2-M phase, with modulation of differentiation markers. Given the potential pivotal role of HuR in regulating cell proliferation and differentiation, we treated keratinocytes with TM molecules able to selectively bind to HuR while modulating its activity. We demonstrated a dose-dependent inhibition of proliferation both in keratinocytes and in T cells, with modulation of Cyclin B, and an increase of apoptosis at higher concentrations. Psoriasis-derived cells appeared to be more sensitive to treatments. Moreover, psoriatic markers, such as CK16 and EFABP, are decreased by treatments together with modulation of proliferation markers, such as Survivin. Finally, treatments appeared to exert also an immunomodulatory function, increasing the frequency of Th1 cells instead of Th17, modulating cytokine production, such as IL17A, IFNg, CXCL8, and activation of Caspase 1. Further analysis of HuR target molecules is now under evaluation by CLIP-seq analysis on psoriasis patients’ samples. Taken together, these data demonstrate that the inhibition of HuR holds promise for an innovative ground-breaking strategy to limit cytokine release and concomitant blockade of keratinocytes-lymphocytes proliferation and cross-talk.
P06 NOS2-derived low levels of NO drive psoriasis pathogenesis
Journal: British Journal of Dermatology (2024)

Abstract Psoriasis is an IL-23/Th17-mediated skin disorder with a strong genetic predisposition. The impact of its susceptibility gene nitric oxide synthase 2 (NOS2) remains unknown. Here, we demonstrate strong NOS2 mRNA expression in psoriatic epidermis, an effect that is IL-17 dependent. However, its complete translation to protein is prevented by the IL-17-induced miR-31 implying marginally upregulated NO levels in psoriatic skin. We demonstrate that lower levels of NO, as opposed to higher levels, increase keratinocyte proliferation and mediate IL-17 downstream effects. We hypothesized that the psoriatic phenotype may be alleviated by either eliminating or increasing cellular NO levels. In fact, using the imiquimod psoriasis mouse model, we found a profound impact on the psoriatic inflammation in both IMQ-treated NOS2 KO mice and wild-type mice treated with IMQ and the NO-releasing berdazimer gel. In conclusion, we demonstrate that IL-17 induces NOS2 and fine-tunes its translation towards a window of proinflammatory and hyperproliferative effects and identify NO donor therapy as a new treatment modality for psoriasis.