Publications

<p>Schematic overview mechanism of Ewing sarcoma cells shaping the immune microenvironment into an immunosuppressive niche. (<a href="#bib1" target="_blank">1</a>) Immunomodulatory interactions between Ewing sarcoma cells and myeloid cells induce polarization of macrophages to an immunosuppressive, M2-like, phenotype and induce tolerogenic DCs. (<a href="#bib2" target="_blank">2</a>) Immunosuppressive macrophage interacts with T cell, where, for example, TIGIT–NECTIN2 interactions leads to the inhibition of T-cell activation resulting in a suppressed/dysfunctional T cell. (<a href="#bib3" target="_blank">3</a>) Tolerogenic DC interacts with naïve T cells but is lacking expression of costimulatory molecules, important for T-cell priming, resulting in a tolerogenic T cell. (<a href="#bib4" target="_blank">4</a>) T cell interacts with Ewing sarcoma cell, but because of it tolerogenic state the T cell is unable to activate, produce cytokines, and proliferate which are essential for cancer cell killing. Image created using BioRender.</p>

<p>Immunosuppressed and functionally impaired myeloid cells in Ewing sarcoma</p>

Well‐being of intensive care nurses (WEBIC): a job analytic approach Aims of the study. This paper presents the results of a validation study of the so‐called well‐being of intensive care nurses (WEBIC)‐questionnaire that is designed to perform a detailed job analysis of intensive care unit (ICU) nurses’ jobs. Background. The WEBIC‐questionnaire is based on modern sociotechnical systems theory, and distinguishes four integrated task categories: (1) operational, (2) organizing, (3) preparatory, and (4) supportive tasks. For each task, the WEBIC assesses (1) how demanding this task is, and (2) how satisfying the performance of this task is. Using the WEBIC, information is gathered about ICU nurses’ qualitative workload, and typical job‐related risks for ICU nurses’ well‐being at work can be mapped. Methods. A cross‐sectional survey on work and well‐being of almost 2000 ICU‐nurses in 13 different European areas was conducted. Exploratory factor analyses were performed to study the validity of the factorial structure of the WEBIC‐questionnaire. The construct validity of the WEBIC‐questionnaire was studied by performing hierarchical multiple regression analyses of the WEBIC‐factors on two types of job‐related well‐being, i.e. burnout and general job satisfaction. Results. Results of the exploratory factor analyses showed that the hypothesized four‐factor structure of the WEBIC is confirmed by our data. Internal consistencies of the different factors varied from 0·77 to 0·91. Intensive care unit nurses’ most central (operational) tasks turned out to pose the greatest demands, but also seemed to drive their satisfaction. With respect to the relationships between the four WEBIC‐factors, and burnout and general job satisfaction, it was found that, especially for the satisfying tasks, significant relationships with these outcomes were found. Conclusion. The reliability and construct validity of the WEBIC‐questionnaire can be considered satisfactory. Furthermore, the questionnaire provides a systematical and detailed coverage of ICU nurses’ tasks. In relation to this, the questionnaire is not only useful for scientific purposes but also for practical use.

Background: Organoids are 3D structures that recapitulate morphological, functional and genetic characteristics of the tissue of origin. Grown as organoids, primary cells can be kept in culture long-term without the requirement for immortalization or genetic alterations. In this study, we report the establishment of organoids derived of Head and Neck Squamous Cell Carcinomas (HNSCCs), which are tumors that arise from the squamous epithelium lining the oral cavity, pharynx and larynx. Overall five-year survival of patients diagnosed with this tumor is poor, and although many of the mutations detected in these tumors are therapeutically targetable, introduction of these treatment into the clinic has failed. Methods: Organoids are established from HNSCC biopsies or surgical resections. Subsequently, the organoids are characterized using histology and DNA and RNA sequencing. In vitro, organoids are tested for sensitivity to a range of drugs. Results: Here we report the establishment of organoids derived of HNSCCs. These structures seem to recapitulate genetic and histological features of the tumors of which they are derived. Upon drug exposure in vitro, these organoids show differential responses to different therapies, including cisplatin, carboplatin and the anti-EGFR antibody cetuximab, which are currently used in the clinic. Our data suggests that this system might have the potential to guide personalized therapy in the future. To validate these findings, we are currently setting up a study to test the predictive potential of patient-derived organoid lines for the first line treatment of HNSCC patients. We will expose tumor-derived organoids to these therapies. Retrospectively, organoid responses will be compared to patient response. The results of this study will show if, in this patient group, organoids can serve as a tool to guide therapy choice. Conclusions: This work suggests that HNSCC-derived organoids might have the potential to predict patient response, a hypothesis we will hope to validate by the study described above. Retrospectively, we will correlation patient outcome to in vitro organoid response. This will help us determine the potential of this system to predict therapy response of patients with HNSCC. Legal entity responsible for the study: Hans Clevers research group, Hubrecht Institute. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.

Abstract Pluripotent stem cell (PSC)-derived human brain organoids enable the study of human brain development in vitro. Typically, the fate of PSCs is guided into subsequent specification steps through static medium switches. In vivo, morphogen gradients are critical for proper brain development and determine cell specification, and associated defects result in neurodevelopmental disorders. Here, we show that initiating neural induction in a temporal stepwise gradient guides the generation of brain organoids composed of a single, self-organized apical-out neuroepithelium, termed ENOs (expanded neuroepithelium organoids). This is at odds with standard brain organoid protocols in which multiple and independent neuroepithelium units (rosettes) are formed. We find that a prolonged, decreasing gradient of TGF-β signaling is a determining factor in ENO formation and allows for an extended phase of neuroepithelium expansion. In-depth characterization reveals that ENOs display improved cellular morphology and tissue architectural features that resemble in vivo human brain development, including expanded germinal zones. Consequently, cortical specification is enhanced in ENOs. ENOs constitute a platform to study the early events of human cortical development and allow interrogation of the complex relationship between tissue architecture and cellular states in shaping the developing human brain.

Abstract Organoids are self-organizing 3D structures grown from stem cells that recapitulate essential aspects of organ structure and function. Here we describe a method to establish long-term-expanding human airway organoids from broncho-alveolar biopsies or lavage material. The pseudostratified airway organoid epithelium consists of basal cells, functional multi-ciliated cells, mucus-producing goblet cells, and CC10-secreting club cells. Airway organoids derived from cystic fibrosis (CF) patients allow assessment of CFTR function in an organoid swelling assay. Organoid culture conditions also allow gene editing as well as the derivation of various types of lung cancer organoids. Respiratory syncytial virus (RSV) infection recapitulated central disease features and dramatically increases organoid cell motility, found to be driven by the non-structural viral NS2 protein. We conclude that human airway organoids represent versatile models for the in vitro study of hereditary, malignant, and infectious pulmonary disease.