Publications

Head and neck (HN) cancer is a broad category of tumor types arising from various anatomic structures including the craniofacial bones, soft tissues, salivary glands, skin, and mucosal membranes. Treatment often involves an intensive combination of surgery, radiotherapy and chemotherapy. Despite this, tumour recurrence rates remain high and survival rates are relatively poor. Here, we describe a novel high throughput drug screening platform combining the OrganoPlate, a microfluidic based 3D culture plate, and HN cancer-derived organoids. MIMETAS develops Organ-on-a-Chip-based models for evaluation of new medicines. Our unique microfluidic technology enables testing of compounds on miniaturized 3D organ models in high-throughput. These models are expected to show better predictivity as compared to laboratory animals and conventional 2D cell culture models, without compromising throughput or ease of use. Hereby we show the establishment of HN cancer-derived organoids in 2-lane OrganoPlate, and its usefulness for phenotypic drug screenings. The aim of the study is to evaluate the 2-lane OrganoPlate as a platform for growing HN cancer organoids and drug screening. Organoid lines (T2, T3 and T4) were embedded in the 2-lane OrganoPlate as single cells in an Extracellular Matrix gel. At day 3, cultures were treated with Cisplatin or Carboplatin for 120 hours. Drug response was evaluated by assessment of morphology (phase contrast), Cell Viability (Alamar blue) and proliferation (EdU incorporation). Organoids cultures grow well under perfusion in the 2-lane OrganoPlate and different sensitivity to cisplatin is captured by the used readouts. The high-throughput, microfluidic 2-lane OrganoPlate platform offers an attractive method for growing HN cancer-derived organoids, supporting development of individualized tumour models for phenotypic drug screenings.Citation Format: Karla Queiroz, Else Driehuis, Silvia Bonilla, Henriëtte Lanz, Hans Clevers, Jos Joore, Paul Vulto. A novel high throughput platform for head & neck cancer organoids drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1173.

Supportive studies of the demand-control (DC) model were more likely to measure specific demands combined with a corresponding aspect of control.

The circumvallate papillae and foliate papillae of the posterior tongue contain taste buds in close proximity to specialized salivary glands, known as von Ebner glands. The developmental relationship between taste buds and these salivary glands has been suggested but remains largely unexplored at postnatal and adult stages. Lineage tracing studies in mice have revealed that Lgr5 marks taste bud stem cells. Here, we report single-cell RNA sequencing of the entire circumvallate and foliate papillae of mice, providing a transcriptional atlas of cells from tongue surface epithelium, taste buds, and the associated and non-associated salivary glands. We unveil a developmental trajectory in which taste buds, the associated salivary glands, and the non-taste tongue surface epithelium originate from a common Lgr5 cell. We confirm this tripotency at the clonal level in vitro and with multicolor lineage tracing in vivo. Thus, the circumvallate and foliate papillae harbor chemosensory units composed of taste bud and salivary gland cells derived from the same parental Lgr5-positive stem cell.

Spirituality is an important theme in health research, since a spiritual orientation can help people to cope with the consequences of a serious disease. Knowledge on the role of spirituality is, however, limited, as most research is based on measures of religiosity rather than spirituality. A questionnaire that transcends specific beliefs is a prerequisite for quantifying the importance of spirituality among people who adhere to a religion or none at all. In this review, we discuss ten questionnaires that address spirituality as a universal human experience. Questionnaires are evaluated with regard to psychometric properties, item formulation and confusion with well-being and distress. Although none of the questionnaires fulfilled all the criteria, the multidimensional Spiritual Well-Being Questionnaire is promising.

Abstract Previous studies have described that tumor organoids can capture the diversity of defined human carcinoma types. Here, we describe conditions for long-term culture of human mucosal organoids. Using this protocol, a panel of 31 head and neck squamous cell carcinoma (HNSCC)–derived organoid lines was established. This panel recapitulates genetic and molecular characteristics previously described for HNSCC. Organoids retain their tumorigenic potential upon xenotransplantation. We observe differential responses to a panel of drugs including cisplatin, carboplatin, cetuximab, and radiotherapy in vitro. Additionally, drug screens reveal selective sensitivity to targeted drugs that are not normally used in the treatment of patients with HNSCC. These observations may inspire a personalized approach to the management of HNSCC and expand the repertoire of HNSCC drugs. Significance: This work describes the culture of organoids derived from HNSCC and corresponding normal epithelium. These tumoroids recapitulate the disease genetically, histologically, and functionally. In vitro drug screening of tumoroids reveals responses to therapies both currently used in the treatment of HNSCC and those not (yet) used in clinical practice. See related commentary by Hill and D'Andrea, p. 828. This article is highlighted in the In This Issue feature, p. 813

This study investigated, among medical specialists ( N = 2,400), the association between perceived inequity in relationships at work (patients, colleagues, organization) and burnout, and the moderating role of communal orientation. Intrapersonal inequity, involving an internal standard of reference, and interpersonal inequity, taking colleagues as the standard of reference, were assessed. The adjusted response rate was 63%. Intrapersonal inequity affected all burnout dimensions: emotional exhaustion in all relationships at work, depersonalization in the relationship with patients, and reduced personal accomplishment in relationships with colleagues and the organization. Intrapersonal inequity explained more variance in burnout than did interpersonal inequity. Communal orientation did not moderate these associations. Hence, medical specialists are more vulnerable to burnout if they perceive their relationships at work to be inequitable, regardless of their tendency to help others. Organizations might, therefore, prevent burnout by creating a working environment that is supportive and appreciative.

The heterogeneous nature of colorectal cancer (CRC) complicates prognosis and is suggested to be a determining factor in the efficacy of adjuvant therapy for individual patients. Based on gene expression profiling, CRC is currently classified into four consensus molecular subtypes (CMSs), characterized by specific biological programs, thus suggesting the existence of unifying developmental drivers for each CMS Using human organoid cultures, we investigated the role of such developmental drivers at the premalignant stage of distinct CRC subtypes and found that TGFβ plays an important role in the development of the mesenchymal CMS4, which is of special interest due to its association with dismal prognosis. We show that in tubular adenomas (TAs), which progress to classical CRCs, the dominating response to TGFβ is death by apoptosis. By contrast, induction of a mesenchymal phenotype upon TGFβ treatment prevails in a genetically engineered organoid culture carrying a BRAF(V) (600E) mutation, constituting a model system for sessile serrated adenomas (SSAs). Our data indicate that TGFβ signaling is already active in SSA precursor lesions and that TGFβ is a critical cue for directing SSAs to the mesenchymal, poor-prognosis CMS4 of CRC.

Patient-derived tumor xenograft (PDX)/organoid (PDO), driven by cancer stem cells (CSC), are considered the most predictive models for translational oncology. Large PDX collections reflective of patient populations have been created and used extensively to test various investigational therapies, including population-trials as surrogate subjects in vivo . PDOs are recognized as in vitro surrogates for patients amenable for high-throughput screening (HTS). We have built a biobank of carcinoma PDX-derived organoids (PDXOs) by converting an existing PDX library and confirmed high degree of similarities between PDXOs and parental PDXs in genomics, histopathology and pharmacology, suggesting “biological equivalence or interchangeability” between the two. Here we demonstrate the applications of PDXO biobank for HTS “matrix” screening for both lead compounds and indications, immune cell co-cultures for immune-therapies and engineering enables in vitro/in vivo imaging. This large biobank of >550 matched pairs of PDXs/PDXOs across different cancers could become powerful tools for the future cancer drug discovery.

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Objective To examine the prevalence, antecedents and consequences of physician care left undone in acute care hospitals. Design A multicentre, multinational, cross-sectional survey. An 11-item scale measured physician reports of care left undone. Antecedent measures examined were work environment and perceived workload. Potential consequences examined included emotional exhaustion, job dissatisfaction and perceived quality of care. Generalized linear mixed models were estimated to quantify associations between physician care left undone and the theorized antecedents and consequences. Setting 56 acute care hospitals in six European countries. Participants 1 963 physicians providing direct patient care to adult in-patients. Results Four in five (78.3 %) physicians left one or more care activities undone during their last shift. On average 3.1 (SD 1.0) of 11 activities were left undone. This varied between and within countries. A 10 % increase at the hospital level of physicians saying they have too much work to do, significantly increased the odds of one or more activities being left undone (OR 1.414, 95 % CI 1.268–1.578). Physicians’ reports of care left undone were associated with increased odds of emotional exhaustion (OR 3.867, 95 %CI 2.683–5.575) and rating quality of medical care as poor or fair (OR 3.395, 95 % CI 2.215–5.204). Conclusion Physicians frequently report leaving some necessary care undone. A shortage of resources compromises physicians’ ability to do their jobs, impacting the quality of care they deliver and their job satisfaction and well-being. Ensuring adequate healthcare personnel resources should be a top priority for hospitals.

Abstract Oncogenic mutations in phenotypically normal tissue are common across adult organs 1,2 . This suggests that multiple events need to converge to drive tumorigenesis and that many processes such as tissue differentiation may protect against carcinogenesis. WNT–β-catenin signalling maintains zonal differentiation during liver homeostasis 3,4 . However, the CTNNB1 oncogene—encoding β-catenin—is also frequently mutated in hepatocellular carcinoma, resulting in aberrant WNT signalling that promotes cell growth 5,6 . Here we investigated the antagonistic interplay between WNT-driven growth and differentiation in zonal hepatocyte populations during liver tumorigenesis. We found that β-catenin mutations co-operate with exogenous MYC expression to drive a proliferative translatome. Differentiation of hepatocytes to an extreme zone 3 fate suppressed this proliferative translatome. Furthermore, a GLUL and Lgr5 -positive perivenous subpopulation of zone 3 hepatocytes were refractory to WNT-induced and MYC-induced tumorigenesis. However, when mutant CTNNB1 and MYC alleles were activated sporadically across the liver lobule, a subset of mutant hepatocytes became proliferative and tumorigenic. These early lesions were characterized by reduced WNT pathway activation and elevated MAPK signalling, which suppresses zone 3 differentiation. The proliferative lesions were also dependent on IGFBP2–mTOR–cyclin D1 pathway signalling, in which inhibition of either IGFBP2 or mTOR suppressed proliferation and tumorigenesis. Therefore, we propose that zonal identity dictates hepatocyte susceptibility to WNT-driven tumorigenesis and that escaping WNT-induced differentiation is essential for liver cancer.