IL-38 Ameliorates Skin Inflammation and Limits IL-17 Production from γδ T Cells

Summary: Interleukin-38 (IL-38) is a cytokine of the IL-1 family with a role in chronic inflammation. However, its main cellular targets and receptors remain obscure. IL-38 is highly expressed in the skin and downregulated in psoriasis patients. We report an investigation in cellular targets of IL-38 during the progression of imiquimod-induced psoriasis. In this model, IL-38 knockout (IL-38 KO) mice show delayed disease resolution with exacerbated IL-17-mediated inflammation, which is reversed by the administration of mature IL-38 or γδ T cell-receptor-blocking antibodies. Mechanistically, X-linked IL-1 receptor accessory protein-like 1 (IL1RAPL1) is upregulated upon γδ T cell activation to feedforward-amplify IL-17 production and is required for IL-38 to suppress γδ T cell IL-17 production. Accordingly, psoriatic IL1RAPL1 KO mice show reduced inflammation and IL-17 production by γδ T cells. Our findings indicate a role for IL-38 in the regulation of γδ T cell activation through IL1RAPL1, with consequences for auto-inflammatory disease. : Han et al. report that genetic depletion of IL-38 in mice delays the resolution of imiquimod-induced psoriasis by increasing the production of the inflammatory cytokine IL-17A by skin-infiltrating T cells. Depleting these T cells or the receptor that is targeted by IL-38 reduces psoriatic skin inflammation. Keywords: IL-38, IL1RAPL1, IL-17, γδ T cells, psoriasis, inflammatio

The fear of the other : a vignette experiment about threat perception : results of a student research seminar.

Dieser Arbeitsbericht fasst die Ergebnisse eines Vignettenexperiments zu Bedrohungswahrnehmungen in Alltagssituationen zusammen, das im Rahmen eines studentischen Forschungsprojekts durchgeführt wurde. Es wird der Frage nachgegangen, ob zugeschriebene äußere Charakteristika (z. B.Ethnizität und Geschlecht) von Fremdgruppen die Bedrohungswahrnehmung beeinflussen und ob diese Einflussbeziehung in Abhängigkeit von Kontextbedingungen oder in Abhängigkeit von Befragtenmerkmalen variiert. Die analysierten Daten basieren auf einer Primärerhebung einer studentischen Stichprobe im Rahmen einer Online-Befragung. Die dabei gewonnenen Daten werden anhand eines statistischen Mehrebenenmodells ausgewertet. Im Ergebnis zeigt sich, dass die aufgestellten Hypothesen bestätigt werden können, wobei jedoch weitere, nicht erhobene Einflussgrößen die Bedrohungswahrnehmung determinieren

Novel conformation specific inhibitors of activated GTPases reveal Ras-dependency of patient-derived cancer organoids

The small GTPases H, K, and NRAS are molecular switches that are indispensable for proper regulation of cellular proliferation and growth. Mutations in this family of proteins are associated with cancer and result in aberrant activation of signaling processes caused by a deregulated recruitment of downstream effector proteins. In this study, we engineered novel variants of the Ras-binding domain (RBD) of the kinase CRAF. These variants bound with high affinity to the effector binding site of active Ras. Structural characterization showed how the newly identified mutations cooperate to enhance affinity to the effector binding site compared to RBDwt. The engineered RBD variants closely mimic the interaction mode of naturally occurring Ras effectors and as dominant negative affinity reagent block their activation. Experiments with cancer cells showed that expression of these RBD variants inhibits Ras signaling leading to a reduced growth and inductions of apoptosis. Using the optimized RBD variants, we stratified patient-derived colorectal cancer organoids according to Ras dependency, which showed that the presence of Ras mutations was insufficient to predict sensitivity to Ras inhibition

Robust, reproducible and quantitative analysis of thousands of proteomes by micro-flow LC-MS/MS

Nano-flow liquid chromatography tandem mass spectrometry (nano-flow LC-MS/MS) is the mainstay in proteome research because of its excellent sensitivity but often comes at the expense of robustness. Here we show that micro-flow LC-MS/MS using a 1 × 150 mm column shows excellent reproducibility of chromatographic retention time (2000 samples of human cell lines, tissues and body fluids. Deep proteome analysis identifies >9000 proteins and >120,000 peptides in 16 h and sample multiplexing using tandem mass tags increases throughput to 11 proteomes in 16 h. The system identifies >30,000 phosphopeptides in 12 h and protein-protein or protein-drug interaction experiments can be analyzed in 20 min per sample. We show that the same column can be used to analyze >7500 samples without apparent loss of performance. This study demonstrates that micro-flow LC-MS/MS is suitable for a broad range of proteomic applications

Circular synthesized CRISPR/Cas gRNAs for functional interrogations in the coding and noncoding genome

Current technologies used to generate CRISPR/Cas gene perturbation reagents are labor intense and require multiple ligation and cloning steps. Furthermore, increasing gRNA sequence diversity negatively affects gRNA distribution, leading to libraries of heterogeneous quality. Here, we present a rapid and cloning-free mutagenesis technology that can efficiently generate covalently-closed-circular-synthesized (3Cs) CRISPR/Cas gRNA reagents and that uncouples sequence diversity from sequence distribution. We demonstrate the fidelity and performance of 3Cs reagents by tailored targeting of all human deubiquitinating enzymes (DUBs) and identify their essentiality for cell fitness. To explore high-content screening, we aimed to generate the largest up-to-date gRNA library that can be used to interrogate the coding and noncoding human genome and simultaneously to identify genes, predicted promoter flanking regions, transcription factors and CTCF binding sites that are linked to doxorubicin resistance. Our 3Cs technology enables fast and robust generation of bias-free gene perturbation libraries with yet unmatched diversities and should be considered an alternative to established technologies

Quality apprenticeships: a manual for educational organisations

The project “Mainstreaming Procedures for Quality Apprenticeships in Educational Organisations and Enterprises” (ApprenticeshipQ) will support educational institutions and placement providers to offer high-quality education to their apprentices. We consider educational institutions here as Higher Vocational Education and Training Institutions, and Professional Higher Education Institutions as Universities of Applied Sciences and Colleges, as well as Academic/ Research-oriented Universities). The project’s proposed assessment of quality will make these processes more manageable and will benefit all stakeholders. These benefits include lifelong learning for teachers, professors and tutors, enhancements for placement providers, improvement of apprentice’s skills development and overall higher quality of the learning experience. The activities of ApprenticeshipQ will strengthen the cooperation and networking between educational institutions and their training partners’ site (placement providers), by providing them with innovative practices to enhance or establish quality management documented information that was developed and tested during the project. The aim is to develop management tools that support educational institutions and placement providers to offer and direct high-quality apprenticeships. These management tools shall be recognised as a model to improve the quality of the apprenticeships and will serve as a basis for the development of formal international standards and guidelines. This guide is structured as follows: The guide starts by giving an introduction to the ApprenticeshipQ project, containing definitions of the research, which led to this guide. Guidance on how to use this guide follows. This section provides a more detailed description of the present document and its objectives. The quality criteria developed in the project are listed with the corresponding measurement indicators in chapter 5. Each criterion is supplemented by a short description and an example. The examples are taken from their project qualitative interview survey with numerous placement providers. Also, a link to documented information is provided for each quality criteria. How these criteria can be implemented is explained in Chapter 6 using a PDCA (PlanDo-Check-Act-Cycle) model

Quality apprenticeships: a manual for placement providers

The project “Mainstreaming Procedures for Quality Apprenticeships in Educational Organisations and Enterprises” (ApprenticeshipQ) will support educational institutions and placement providers to offer high-quality education to their apprentices. We consider educational institutions here as Higher Vocational Education and Training Institutions, and Professional Higher Education Institutions as Universities of Applied Sciences and Colleges, as well as Academic/ Research-oriented Universities). The project’s proposed assessment of quality will make these processes more manageable and will benefit all stakeholders. These benefits include lifelong learning for teachers, professors and tutors, enhancements for placement providers, improvement of apprentice’s skills development and overall higher quality of the learning experience. The activities of ApprenticeshipQ will strengthen the cooperation and networking between educational institutions and their training partners’ site (placement providers), by providing them with innovative practices to enhance or establish quality management documented information that was developed and tested during the project. The aim is to develop management tools that support educational institutions and placement providers to offer and direct high-quality apprenticeships. These management tools shall be recognised as a model to improve the quality of the apprenticeships and will serve as a basis for the development of formal international standards and guidelines. This guide is structured as follows: The guide starts by giving an introduction to the ApprenticeshipQ project, containing definitions of the research, which led to this guide. Guidance on how to use this guide follows. This section provides a more detailed description of the present document and its objectives. The quality criteria developed in the project are listed with the corresponding measurement indicators in chapter 5. Each criterion is supplemented by a short description and an example. The examples are taken from their own project qualitative interview survey with numerous placement providers. Also, a link to documented information is provided for each quality criteria. How these criteria can be implemented is explained in Chapter 6 using a PDCA (PlanDo-Check-Act-Cycle) model

Inhibiting Hippo pathway kinases releases WWC1 to promote AMPAR-dependent synaptic plasticity and long-term memory in mice

The localization, number, and function of postsynaptic AMPA-type glutamate receptors (AMPARs) are crucial for synaptic plasticity, a cellular correlate for learning and memory. The Hippo pathway member WWC1 is an important component of AMPAR-containing protein complexes. However, the availability of WWC1 is constrained by its interaction with the Hippo pathway kinases LATS1 and LATS2 (LATS1/2). Here, we explored the biochemical regulation of this interaction and found that it is pharmacologically targetable in vivo. In primary hippocampal neurons, phosphorylation of LATS1/2 by the upstream kinases MST1 and MST2 (MST1/2) enhanced the interaction between WWC1 and LATS1/2, which sequestered WWC1. Pharmacologically inhibiting MST1/2 in male mice and in human brain-derived organoids promoted the dissociation of WWC1 from LATS1/2, leading to an increase in WWC1 in AMPAR-containing complexes. MST1/2 inhibition enhanced synaptic transmission in mouse hippocampal brain slices and improved cognition in healthy male mice and in male mouse models of Alzheimer's disease and aging. Thus, compounds that disrupt the interaction between WWC1 and LATS1/2 might be explored for development as cognitive enhancers.</p