Publications

<p>Cancer-associated fibroblast subsets in Ewing sarcoma</p>

Patient-derived organoid (PDO) models allow for long-term expansion and maintenance of primary epithelial cells grown in three dimensions and a near-native state. When derived from resected or biopsied tumor tissue, organoids closely recapitulate in vivo tumor morphology and can be used to study therapy response in vitro. Biobanks of tumor organoids reflect the vast variety of clinical tumors and patients and therefore hold great promise for preclinical and clinical applications. This paper presents a method for medium-throughput drug screening using head and neck squamous cell carcinoma and colorectal adenocarcinoma organoids. This approach can easily be adopted for use with any tissue-derived organoid model, both normal and diseased. Methods are described for in vitro exposure of organoids to chemo- and radiotherapy (either as single-treatment modality or in combination). Cell survival after 5 days of drug exposure is assessed by measuring adenosine triphosphate (ATP) levels. Drug sensitivity is measured by the half-maximal inhibitory concentration (IC50), area under the curve (AUC), and growth rate (GR) metrics. These parameters can provide insight into whether an organoid culture is deemed sensitive or resistant to a particular treatment.

<p>Ewing sarcoma–associated lymphocytes display variable degrees of dysfunctionality and immune suppression. <b>A,</b> UMAP and dotplot of the T- and NK-cell subsets and a selection of their marker genes. <b>B,</b> Heat map showing average expression of inhibitory receptors in the T- and NK-cell subsets. *Significantly upregulated with <i>P</i> < 0.0001 using Wilcoxon rank-sum test with Bonferroni correction. <b>C,</b> Dysfunction scores constructed using genes from B. Cell types ordered from high to low score. *, <i>P</i> < 0.05; ***, <i>P</i> < 0.001 versus Naïve T.</p>

Spatiotemporal control of Wnt signaling is essential for the development and homeostasis of many tissues. The transmembrane E3 ubiquitin ligases ZNRF3 (zinc and ring finger 3) and RNF43 (ring finger protein 43) antagonize Wnt signaling by promoting degradation of frizzled receptors. ZNRF3 and RNF43 are frequently inactivated in human cancer, but the molecular and therapeutic implications remain unclear. Here, we demonstrate that adrenocortical-specific loss of ZNRF3, but not RNF43, results in adrenal hyperplasia that depends on Porcupine-mediated Wnt ligand secretion. Furthermore, we discovered a Wnt/β-catenin signaling gradient in the adrenal cortex that is disrupted upon loss of ZNRF3. Unlike β-catenin gain-of-function models, which induce high Wnt/β-catenin activation and expansion of the peripheral cortex, ZNRF3 loss triggers activation of moderate-level Wnt/β-catenin signaling that drives proliferative expansion of only the histologically and functionally distinct inner cortex. Genetically reducing β-catenin dosage significantly reverses the ZNRF3-deficient phenotype. Thus, homeostatic maintenance of the adrenal cortex is dependent on varying levels of Wnt/β-catenin activation, which is regulated by ZNRF3.

Investigation of diseases of the bile duct system and identification of potential therapeutic targets are hampered by the lack of tractable in vitro systems to model cholangiocyte biology. Here, we show a step-wise method for the differentiation of murine Lgr5⁺ liver stem cells (organoids) into cholangiocyte-like cells (CLCs) using a combination of growth factors and extracellular matrix components. Organoid-derived CLCs display key properties of primary cholangiocytes, such as expressing cholangiocyte markers, forming primary cilia, transporting small molecules and responding to farnesoid X receptor agonist. Integration of organoid-derived cholangiocytes with collagen-coated polyethersulfone hollow fiber membranes yielded bioengineered bile ducts that morphologically resembled native bile ducts and possessed polarized bile acid transport activity. As such, we present a novel in vitro model for studying and therapeutically modulating cholangiocyte function.

<p>Immunoregulatory interactions shape the immunosuppressive microenvironment of Ewing sarcoma. <b>A,</b> Number of unique interactions between tumor cells and indicated main cell types. <b>B,</b> Bubble plot of predicted immunosuppressive interactions between indicated myeloid subsets and tumor cells. For full overview see <a href="#SMF9" target="_blank">Supplementary Fig. S9A</a>. <b>C,</b> Dotplot of genes included in the selected, predicted interactions, shown in B, and expressed by the main cell types and per-patient tumor cells. <b>D,</b> Number of unique interactions between T/NK cells and indicated main cell types. <b>E,</b> Bubble plot of predicted interactions between indicated myeloid subsets and CD8<sup>+</sup> T cells and are correlating (>0.5) with CD8<sup>+</sup> T-cell dysfunction score. <b>F,</b> Dotplot of genes included in T cell dysfunction–promoting interactions, highlighted in orange in E, and expressed by the myeloid cells.</p>

Work engagement and workaholism. The role of psychological need satisfaction and consequences for performance Work engagement and workaholism. The role of psychological need satisfaction and consequences for performance Based on Deci and Ryan’s Self-Determination Theory, this study examined possible antecedents of work engagement and workaholism. Furthermore, it examined how these two types of heavy work investment relate to different aspects of job performance (in-role and extra-role performance, counterproductive behavior, and turnover intention). In total 275 health care employees in the Netherlands filled out an online questionnaire. Structural Equation Modeling showed that satisfaction of the psychological need for autonomy and relatedness was associated with work engagement, whereas failure to satisfy the psychological need for autonomy and competence was associated with workaholism. Furthermore, these two types of heavy work investment were positively related to extra-role behavior, and work engagement was negatively related to turnover intention. In addition, the analyses revealed that need satisfaction showed direct relationships with different aspects of job performance. This study offers practical suggestions for promoting work engagement and reducing workaholism, and for optimizing job performance.

<p>Movie S1</p>

Introduction The increasing burden of mental distress reported by healthcare professionals is a matter of serious concern and there is a growing recognition of the role of the workplace in creating this problem. Magnet hospitals, a model shown to attract and retain staff in US research, creates positive work environments that aim to support the well-being of healthcare professionals. Methods and analysis Magnet4Europe is a cluster randomised controlled trial, with wait list controls, designed to evaluate the effects of organisational redesign, based on the Magnet model, on nurses’ and physicians’ well-being in general acute care hospitals, using a multicomponent implementation strategy. The study will be conducted in more than 60 general acute care hospitals in Belgium, England, Germany, Ireland, Norway and Sweden. The primary outcome is burnout among nurses and physicians, assessed in longitudinal surveys of nurses and physicians at participating hospitals. Additional data will be collected from them on perceived work environments, patient safety and patient quality of care and will be triangulated with data from medical records, including case mix-adjusted in-hospital mortality. The process of implementation will be evaluated using qualitative data from focus group and key informant interviews. Ethics and dissemination This study was approved by the Ethics Committee Research UZ/KU Leuven, Belgium; additionally, ethics approval is obtained in all other participating countries either through a central or decentral authority. Findings will be disseminated at conferences, through peer-reviewed manuscripts and via social media. Trial registration number ISRCTN10196901 .