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Enteroendocrine cells (EECs) secrete hormones in response to ingested nutrients to control physiological processes such as appetite and insulin release. EEC hormones are synthesized as large proproteins that undergo proteolytic processing to generate bioactive peptides. Mutations in EEC-enriched proteases are associated with endocrinopathies. Due to the relative rarity of EECs and a paucity of in vitro models, intestinal prohormone processing remains challenging to assess. Here, human gut organoids in which EECs can efficiently be induced are subjected to CRISPR-Cas9–mediated modification of EEC-expressed endopeptidase and exopeptidase genes. We employ mass spectrometry–based analyses to monitor peptide processing and identify glucagon production in intestinal EECs, stimulated upon bone morphogenic protein (BMP) signaling. We map the substrates and products of major EECs endo- and exopeptidases. Our studies provide a comprehensive description of peptide hormones produced by human EECs and define the roles of specific proteases in their generation.

<p>PDF file - 131KB, Wnt target genes and probesets (S1). ER target genes and probesets (S2). Used datasets (S3). Variation in predicted probability of pathway activity between and within cell lines, assessed on dataset E-MTAB-37 (S4). Gene set enrichment analysis results on normal colon samples versus colon adenoma from GSE8671 (S5).</p>

Tumor organoids from individual patients with metastatic colorectal cancer can help predict response to irinotecan with or without 5-fluorouracil.

<p>Summary of clinical features for patient derived xenograft and patient derived organoids</p>

Abstract The biology of cancer is characterized by an intricate interplay of cells originating not only from the tumor mass, but also its surrounding environment. Different microbial species have been suggested to be enriched in tumors and the impacts of these on tumor phenotypes is subject to intensive investigation. For these efforts, model systems that accurately reflect human–microbe interactions are rapidly gaining importance. Here we present a guide for selecting a suitable in vitro co‐culture platform used to model different cancer–microbiome interactions. Our discussion spans a variety of in vitro models, including 2D cultures, tumor spheroids, organoids, and organ‐on‐a‐chip platforms, where we delineate their respective advantages, limitations, and applicability in cancer microbiome research. Particular focus is placed on methodologies that facilitate the exposure of cancer cells to microbes, such as organoid microinjections and co‐culture on microfluidic devices. We highlight studies offering critical insights into possible cancer–microbe interactions and underscore the importance of in vitro models in those discoveries. We anticipate the integration of more complex microbial communities and the inclusion of immune cells into co‐culture systems to more accurately simulate the tumor microenvironment. The advent of ever more sophisticated co‐culture models will aid in unraveling the mechanisms of cancer‐microbiome interplay and contribute to exploiting their potential in novel diagnostic and therapeutic strategies.

<p>PDF file - 674KB, Experimental design for the sorting of stem cells and TA cells from the duodenum of Lgr5-GFP-IRES-CreERT mice (Lgr5 Cre+ Catnb+/lox(ex3) Klf5flox/flox, Lgr5 Cre+ Catnb+/lox(ex3) Klf5+/+, and Lgr5 Cre+ Klf5flox/flox) for mRNA expression microarray analysis.</p>

Abstract Pseudomonas aeruginosa is a Gram-negative bacterium that is notorious for infections in the airway of cystic fibrosis (CF) subjects. Often, these infections become chronic, leading to higher morbidity and mortality rates. Bacterial quorum sensing (QS) coordinates the expression of virulence factors and the formation of biofilms at a population level. QS has become the focus of attention for development of alternatives to antimicrobials targeting P. aeruginosa infections. However, a better understanding of the bacteria-host interaction, and the role of QS in infection, is required. In this study, we set up a new P. aeruginosa infection model, using 2D airway organoids derived from healthy and CF individuals. Using dual RNA-sequencing, we dissected their interaction, focusing on the role of QS. As expected, P. aeruginosa induced epithelial inflammation. However, QS signaling did not affect the epithelial airway cells. The epithelium influenced several infection-related processes of P. aeruginosa , including metabolic changes, induction of type 3 and type 6 secretion systems (T3SS and T6SS), and increased expression of antibiotic resistance genes, including mexXY efflux pump and several porins. Interestingly, the epithelium influenced the regulation by QS of the type 2 (T2SS) and T6SS. Finally, we compared our model with in vivo P. aeruginosa transcriptomic datasets, from samples directly isolated from the airways of CF subjects. This shows that our model recapitulates important aspects of in vivo infection, like enhanced denitrification, betaine/choline metabolism, increased antibiotic resistance, as well as an overall decrease of motility-related genes. This relevant infection model is interesting for future investigations, helping to reduce the burden of P. aeruginosa infections in CF.

There is an increasing need for managers to understand what motivates younger versus older workers to continue work within their company. We believe that this two-wave study among 90 Dutch employees is the first to examine: (1) the cross-lagged relationships between breach of psychological contract (which includes transactional and relational obligations) and intrinsic work motivation, and (2) the moderating role of the age-related variables future time perspective and regulatory focus. Regulatory focus concerns the orientation (either promotion-focused or prevention-focused) by which an individual pursues their goals. Based on psychological contract theory, we expected and found that relational contract breach predicts lower work motivation. Furthermore, based on lifespan developmental and regulatory focus theory, we assumed that this relationship would be stronger when workers experienced an open future time perspective and a promotion focus rather than a prevention focus. The results showed that future time perspective indeed had a strengthening, and prevention focus a reducing moderating effect in the relationship between psychological contract breach and work motivation. However, no significant effects for promotion focus were found. These findings indicate that age-related processes such as future time perspective and regulatory focus are important variables to include in future psychological contract research. Keywords: longitudinal researchpsychological contractcontract breachtime perspectiveregulatory focuswork motivationage Acknowledgements The authors would like to thank Ellen Wermelink and Jørgen Nielsen for their help with the data collection.