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Current Biology : CB Jun 2014
Topics: Crohn Disease; Humans; Intestine, Small; Paneth Cells
PubMed: 24937274
DOI: 10.1016/j.cub.2014.04.049 -
Cell Reports Apr 2021Organoids allow the recapitulation of intestinal homeostasis and cancerogenesis in vitro; however, RNA sequencing (RNA-seq)-based methods for drug screens are missing....
Organoids allow the recapitulation of intestinal homeostasis and cancerogenesis in vitro; however, RNA sequencing (RNA-seq)-based methods for drug screens are missing. We develop targeted organoid sequencing (TORNADO-seq), a high-throughput, high-content drug discovery platform that uses targeted RNA-seq to monitor the expression of large gene signatures for the detailed evaluation of cellular phenotypes in organoids. TORNADO-seq is a fast, highly reproducible time- and cost-effective ($5 per sample) method that can probe cell mixtures and their differentiation state in the intestinal system. We apply this method to isolate drugs that enrich for differentiated cell phenotypes and show that these drugs are highly efficacious against cancer compared to wild-type organoids. Furthermore, TORNADO-seq facilitates in-depth insight into the mode of action of these drugs. Our technology can easily be adapted to many other systems and will allow for more systematic, large-scale, and quantitative approaches to study the biology of complex cellular systems.
Topics: Antineoplastic Agents; Cell Differentiation; Colorectal Neoplasms; Drug Discovery; Drug Repositioning; Early Detection of Cancer; Enterocytes; Gene Expression Regulation, Neoplastic; Gene Regulatory Networks; Goblet Cells; High-Throughput Screening Assays; Humans; Neoplasm Proteins; Neoplastic Stem Cells; Organoids; Paneth Cells; Prescription Drugs; RNA-Seq; Sequence Analysis, RNA; Small Molecule Libraries
PubMed: 33882314
DOI: 10.1016/j.celrep.2021.109026 -
Proceedings of the National Academy of... Jan 2022Paneth cells are intestinal epithelial cells that release antimicrobial peptides, such as α-defensin as part of host defense. Together with mesenchymal cells, Paneth...
Paneth cells are intestinal epithelial cells that release antimicrobial peptides, such as α-defensin as part of host defense. Together with mesenchymal cells, Paneth cells provide niche factors for epithelial stem cell homeostasis. Here, we report two subtypes of murine Paneth cells, differentiated by their production and utilization of fucosyltransferase 2 (Fut2), which regulates α(1,2)fucosylation to create cohabitation niches for commensal bacteria and prevent invasion of the intestine by pathogenic bacteria. The majority of Fut2 Paneth cells were localized in the duodenum, whereas the majority of Fut2 Paneth cells were in the ileum. Fut2 Paneth cells showed higher granularity and structural complexity than did Fut2 Paneth cells, suggesting that Fut2 Paneth cells are involved in host defense. Signaling by the commensal bacteria, together with interleukin 22 (IL-22), induced the development of Fut2 Paneth cells. IL-22 was found to affect the α-defensin secretion system via modulation of expression, and IL-17a was found to increase the production of α-defensin in the intestinal tract. Thus, these intestinal cytokines regulate the development and function of Fut2 Paneth cells as part of gut defense.
Topics: Animals; Cytokines; Fucosyltransferases; Gastrointestinal Microbiome; Ileum; Interleukin-17; Interleukins; Mice; Paneth Cells; Symbiosis; alpha-Defensins; Interleukin-22; Galactoside 2-alpha-L-fucosyltransferase
PubMed: 35027453
DOI: 10.1073/pnas.2115230119 -
Viruses Apr 2018Paneth cells are major secretory cells located in the crypts of Lieberkühn in the small intestine. Our understanding of the diverse roles that Paneth cells play in... (Review)
Review
Paneth cells are major secretory cells located in the crypts of Lieberkühn in the small intestine. Our understanding of the diverse roles that Paneth cells play in homeostasis and disease has grown substantially since their discovery over a hundred years ago. Classically, Paneth cells have been characterized as a significant source of antimicrobial peptides and proteins important in host defense and shaping the composition of the commensal microbiota. More recently, Paneth cells have been shown to supply key developmental and homeostatic signals to intestinal stem cells in the crypt base. Paneth cell dysfunction leading to dysbiosis and a compromised epithelial barrier have been implicated in the etiology of Crohn’s disease and susceptibility to enteric bacterial infection. Our understanding of the impact of Paneth cells on viral infection is incomplete. Enteric α-defensins, produced by Paneth cells, can directly alter viral infection. In addition, α-defensins and other antimicrobial Paneth cell products may modulate viral infection indirectly by impacting the microbiome. Here, we discuss recent insights into Paneth cell biology, models to study their function, and the impact, both direct and indirect, of Paneth cells on enteric viral infection.
Topics: Animals; Antimicrobial Cationic Peptides; Humans; Intestinal Diseases; Intestinal Mucosa; Microbiota; Organoids; Paneth Cells; Protein Processing, Post-Translational; Virus Diseases
PubMed: 29701691
DOI: 10.3390/v10050225 -
Cell Sep 2008Inflammatory bowel disease (IBD) has been attributed to aberrant mucosal immunity to the intestinal microbiota. The transcription factor XBP1, a key component of the...
Inflammatory bowel disease (IBD) has been attributed to aberrant mucosal immunity to the intestinal microbiota. The transcription factor XBP1, a key component of the endoplasmic reticulum (ER) stress response, is required for development and maintenance of secretory cells and linked to JNK activation. We hypothesized that a stressful environmental milieu in a rapidly proliferating tissue might instigate a proinflammatory response. We report that Xbp1 deletion in intestinal epithelial cells (IECs) results in spontaneous enteritis and increased susceptibility to induced colitis secondary to both Paneth cell dysfunction and an epithelium that is overly reactive to inducers of IBD such as bacterial products (flagellin) and TNFalpha. An association of XBP1 variants with both forms of human IBD (Crohn's disease and ulcerative colitis) was identified and replicated (rs35873774; p value 1.6 x 10(-5)) with novel, private hypomorphic variants identified as susceptibility factors. Hence, intestinal inflammation can originate solely from XBP1 abnormalities in IECs, thus linking cell-specific ER stress to the induction of organ-specific inflammation.
Topics: Animals; Apoptosis; Colitis, Ulcerative; Crohn Disease; DNA-Binding Proteins; Endoplasmic Reticulum; Genetic Predisposition to Disease; Humans; Inflammatory Bowel Diseases; Intestinal Mucosa; Listeria monocytogenes; Mice; Mice, Transgenic; Paneth Cells; Regulatory Factor X Transcription Factors; Transcription Factors; X-Box Binding Protein 1
PubMed: 18775308
DOI: 10.1016/j.cell.2008.07.021 -
Nature Methods Mar 2020Despite the widespread adoption of organoids as biomimetic tissue models, methods to comprehensively analyze cell-type-specific post-translational modification (PTM)...
Despite the widespread adoption of organoids as biomimetic tissue models, methods to comprehensively analyze cell-type-specific post-translational modification (PTM) signaling networks in organoids are absent. Here, we report multivariate single-cell analysis of such networks in organoids and organoid cocultures. Simultaneous analysis by mass cytometry of 28 PTMs in >1 million single cells derived from small intestinal organoids reveals cell-type- and cell-state-specific signaling networks in stem, Paneth, enteroendocrine, tuft and goblet cells, as well as enterocytes. Integrating single-cell PTM analysis with thiol-reactive organoid barcoding in situ (TOBis) enables high-throughput comparison of signaling networks between organoid cultures. Cell-type-specific PTM analysis of colorectal cancer organoid cocultures reveals that shApc, Kras and Trp53 cell-autonomously mimic signaling states normally induced by stromal fibroblasts and macrophages. These results demonstrate how standard mass cytometry workflows can be modified to perform high-throughput multivariate cell-type-specific signaling analysis of healthy and cancerous organoids.
Topics: Animals; Biomimetics; Cell Differentiation; Coculture Techniques; Colorectal Neoplasms; Cytophotometry; Enterocytes; Enteroendocrine Cells; Female; Fibroblasts; Gene Expression Regulation; Goblet Cells; Humans; Intestine, Small; Macrophages; Mice; Mice, Inbred C57BL; Organ Culture Techniques; Organoids; Paneth Cells; Signal Transduction; Single-Cell Analysis; Sulfhydryl Compounds; Tumor Suppressor Protein p53
PubMed: 32066960
DOI: 10.1038/s41592-020-0737-8 -
Proceedings of the National Academy of... Jun 2023Gut microbiota imbalance (dysbiosis) is increasingly associated with pathological conditions, both within and outside the gastrointestinal tract. Intestinal Paneth cells...
Gut microbiota imbalance (dysbiosis) is increasingly associated with pathological conditions, both within and outside the gastrointestinal tract. Intestinal Paneth cells are considered to be guardians of the gut microbiota, but the events linking Paneth cell dysfunction with dysbiosis remain unclear. We report a three-step mechanism for dysbiosis initiation. Initial alterations in Paneth cells, as frequently observed in obese and inflammatorybowel diseases patients, cause a mild remodeling of microbiota, with amplification of succinate-producing species. SucnR1-dependent activation of epithelial tuft cells triggers a type 2 immune response that, in turn, aggravates the Paneth cell defaults, promoting dysbiosis and chronic inflammation. We thus reveal a function of tuft cells in promoting dysbiosis following Paneth cell deficiency and an unappreciated essential role of Paneth cells in maintaining a balanced microbiota to prevent inappropriate activation of tuft cells and deleterious dysbiosis. This succinate-tuft cell inflammation circuit may also contribute to the chronic dysbiosis observed in patients.
Topics: Humans; Dysbiosis; Mucous Membrane; Inflammation; Paneth Cells; Succinates; Succinic Acid
PubMed: 37307458
DOI: 10.1073/pnas.2219431120 -
EMBO Molecular Medicine Feb 2023Paneth cells are versatile secretory cells located in the crypts of Lieberkühn of the small intestine. In normal conditions, they function as the cornerstones of... (Review)
Review
Paneth cells are versatile secretory cells located in the crypts of Lieberkühn of the small intestine. In normal conditions, they function as the cornerstones of intestinal health by preserving homeostasis. They perform this function by providing niche factors to the intestinal stem cell compartment, regulating the composition of the microbiome through the production and secretion of antimicrobial peptides, performing phagocytosis and efferocytosis, taking up heavy metals, and preserving barrier integrity. Disturbances in one or more of these functions can lead to intestinal as well as systemic inflammatory and infectious diseases. This review discusses the multiple functions of Paneth cells, and the mechanisms and consequences of Paneth cell dysfunction. It also provides an overview of the tools available for studying Paneth cells.
Topics: Paneth Cells; Intestines; Intestine, Small; Microbiota
PubMed: 36573340
DOI: 10.15252/emmm.202216427 -
Gut Microbes 2022Antimicrobial proteins possess a broad spectrum of bactericidal activity and play an important role in shaping the composition of gut microbiota, which is related to...
Antimicrobial proteins possess a broad spectrum of bactericidal activity and play an important role in shaping the composition of gut microbiota, which is related to multiple diseases such as metabolic syndrome. However, it is incompletely known for the regulation of defensin expression in the gut Paneth cells. Here, we found that FABP4 in the Paneth cells of gut epithelial cells and organoids can downregulate the expression of defensins. FABP4pvillin mice were highly resistance to Typhimurium (.T) infection and had increased bactericidal ability to pathogens. The FABP4-mediated downregulation of defensins is through degrading PPARγ after K48 ubiquitination. We also demonstrate that high-fat diet (HFD)-mediated downregulation of defensins is through inducing a robust FABP4 in Paneth cells. / (F/B) ratio in FABP4pvillin mice is lower than control mice, which is opposite to that in mice fed HFD, indicating that FABP4 in the Paneth cells could reprogram gut microbiota. Interestingly, FABP4-mediated downregulation of defensins in Paneth cells not only happens in mice but also in human. A better understanding of the regulation of defensins, especially HFD-mediated downregulation of defensin in Paneth cells will provide insights into factor(s) underlying modern diseases.: FABP4: Fatty acid binding protein 4; S. T: Salmonella Typhimurium; HFD: High-fat diet; Defa: α-defensin; 930 HD5: Human α-defensin 5; HD6: Human α-defensin 6; F/B: Firmicutes/Bacteroidetes; SFB: Segmental filamentous bacteria; AMPs: Antimicrobial peptides; PPARγ: Peroxisome proliferator-activated receptor γ; P-PPAR: Phosphorylated PPAR; Dhx15: DEAD-box helicase 15; 935 EGF: Epidermal growth factor; ENR: Noggin and R-spondin 1; CFU: Colony forming unit; Lyz1: Lysozyme 1; Saa1: Serum amyoid A 1; Pla2g2a: Phospholipase A2, group IIA; MMP-7: Matrix metalloproteinase; AU-PAGE: Acid-urea polyacrylamide gel electrophoresis; PA: Palmitic 940 acid; GPR40: G-protein-coupled receptor; GF: Germ-free; EGF: Epidermal growth factor; LP: Lamina propria; KO: Knock out; WT: Wild-type.
Topics: Animals; Humans; Mice; Anti-Infective Agents; Fatty Acid-Binding Proteins; Gastrointestinal Microbiome; Paneth Cells; RNA Helicases
PubMed: 36519446
DOI: 10.1080/19490976.2022.2139978 -
Gut Microbes 2012Necrotizing enterocolitis (NEC) is a common and devastating disease of premature infants. Immaturity of the innate immune system of the gut is central to the... (Review)
Review
Necrotizing enterocolitis (NEC) is a common and devastating disease of premature infants. Immaturity of the innate immune system of the gut is central to the pathogenesis of NEC. Recent studies suggest a key role for Paneth cells in this disease. Addressing basic questions on the development and function of immature Paneth cells may shed light on the puzzling pathophysiology of NEC. Current animal models of NEC are limited in their capacity to answer these questions.
Topics: Animals; Disease Models, Animal; Enterocolitis, Necrotizing; Humans; Infant, Newborn; Infant, Premature; Intestinal Mucosa; Paneth Cells
PubMed: 22895084
DOI: 10.4161/gmic.21738