P094 PROINFLAMMATORY RESPONSES OF THE INTESTINAL EPITHELIUM ARE PREDOMINANTLY FACILITATED BY STEM CELLS

Graft-versus-host disease (GVHD) is a major complication of allogeneic bone marrow transplantation (BMT), and damage to the gastrointestinal (GI) tract plays a critical role in amplifying systemic disease. Intestinal stem cells (ISCs) play a pivotal role not only in physiological tissue renewal but also in regeneration of the intestinal epithelium after injury. In this study, we have discovered that pretransplant conditioning regimen damaged ISCs; however, the ISCs rapidly recovered and restored the normal architecture of the intestine. ISCs are targets of GVHD, and this process of ISC recovery was markedly inhibited with the development of GVHD. Injection of Wnt agonist R-spondin1 (R-Spo1) protected against ISC damage, enhanced restoration of injured intestinal epithelium, and inhibited subsequent inflammatory cytokine cascades. R-Spo1 ameliorated systemic GVHD after allogeneic BMT by a mechanism dependent on repair of conditioning-induced GI tract injury. Our results demonstrate for the first time that ISC damage plays a central role in amplifying systemic GVHD; therefore, we propose ISC protection by R-Spo1 as a novel strategy to improve the outcome of allogeneic BMT.

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The Hippo–YAP Signaling as Guardian in the Pool of Intestinal Stem Cells

Biomedicines ◽

10.3390/biomedicines8120560 ◽

2020 ◽

Vol 8 (12) ◽

pp. 560

Author(s):

Yoojin Seo ◽

So-Yeon Park ◽

Hyung-Sik Kim ◽

Jeong-Seok Nam

Keyword(s):

Stem Cells ◽

Intestinal Epithelium ◽

Fine Tuning ◽

Intestinal Stem Cells ◽

Dynamic Regulation ◽

Intestinal Integrity ◽

Differentiated Cells ◽

Quiescent Stem Cells ◽

G Protein Coupled ◽

Crypt Base

Despite endogenous insults such as mechanical stress and danger signals derived from the microbiome, the intestine can maintain its homeostatic condition through continuous self-renewal of the crypt–villus axis. This extraordinarily rapid turnover of intestinal epithelium, known to be 3 to 5 days, can be achieved by dynamic regulation of intestinal stem cells (ISCs). The crypt base-located leucine-rich repeat-containing G-protein-coupled receptor 5-positive (Lgr5+) ISCs maintain intestinal integrity in the steady state. Under severe damage leading to the loss of conventional ISCs, quiescent stem cells and even differentiated cells can be reactivated into stem-cell-like cells with multi-potency and contribute to the reconstruction of the intestinal epithelium. This process requires fine-tuning of the various signaling pathways, including the Hippo–YAP system. In this review, we summarize recent advances in understanding the correlation between Hippo–YAP signaling and intestinal homeostasis, repair, and tumorigenesis, focusing specifically on ISC regulation.

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DCAMKL-1 Expression Identifies Tuft Cells Rather Than Stem Cells in the Adult Mouse Intestinal Epithelium

Gastroenterology ◽

10.1053/j.gastro.2009.06.072 ◽

2009 ◽

Vol 137 (6) ◽

pp. 2179-2180 ◽

Cited By ~ 100

Author(s):

François Gerbe ◽

Bénédicte Brulin ◽

Leila Makrini ◽

Catherine Legraverend ◽

Philippe Jay

Keyword(s):

Stem Cells ◽

Intestinal Epithelium ◽

Adult Mouse ◽

Tuft Cells

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Wnt/β-Catenin Is Essential for Intestinal Homeostasis and Maintenance of Intestinal Stem Cells

Molecular and Cellular Biology ◽

10.1128/mcb.01034-07 ◽

2007 ◽

Vol 27 (21) ◽

pp. 7551-7559 ◽

Cited By ~ 370

Author(s):

Tea Fevr ◽

Sylvie Robine ◽

Daniel Louvard ◽

Joerg Huelsken

Keyword(s):

Stem Cells ◽

Wnt Signaling ◽

Intestinal Epithelium ◽

Transcriptional Profiling ◽

Wnt Signaling Pathway ◽

Intestinal Stem Cells ◽

Intestinal Homeostasis ◽

Adult Mice ◽

Gene Ablation ◽

Functional Preservation

ABSTRACT The Wnt signaling pathway is deregulated in over 90% of human colorectal cancers. β-Catenin, the central signal transducer of the Wnt pathway, can directly modulate gene expression by interacting with transcription factors of the TCF/LEF family. In the present study we investigate the role of Wnt signaling in the homeostasis of intestinal epithelium by using tissue-specific, inducible β-catenin gene ablation in adult mice. Block of Wnt/β-catenin signaling resulted in rapid loss of transient-amplifying cells and crypt structures. Importantly, intestinal stem cells were induced to terminally differentiate upon deletion of β-catenin, resulting in a complete block of intestinal homeostasis and fatal loss of intestinal function. Transcriptional profiling of mutant crypt mRNA isolated by laser capture microdissection confirmed those observations and allowed us to identify genes potentially responsible for the functional preservation of intestinal stem cells. Our data demonstrate an essential requirement of Wnt/β-catenin signaling for the maintenance of the intestinal epithelium in the adult organism. This challenges attempts to target aberrant Wnt signaling as a new therapeutic strategy to treat colorectal cancer.

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Porcine Intestinal Enteroids: a New Model for Studying Enteric Coronavirus Porcine Epidemic Diarrhea Virus Infection and the Host Innate Response

Journal of Virology ◽

10.1128/jvi.01682-18 ◽

2018 ◽

Vol 93 (5) ◽

Cited By ~ 12

Author(s):

Liang Li ◽

Fang Fu ◽

Shanshan Guo ◽

Hongfeng Wang ◽

Xijun He ◽

...

Keyword(s):

Stem Cells ◽

Intestinal Epithelium ◽

Porcine Epidemic Diarrhea Virus ◽

Enteric Viruses ◽

Antiviral Response ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea ◽

Ifn Lambda

ABSTRACTPorcine epidemic diarrhea virus (PEDV), a member of the group of alphacoronaviruses, is the pathogen of a highly contagious gastrointestinal swine disease. The elucidation of the events associated with the intestinal epithelial response to PEDV infection has been limited by the absence of goodin vitroporcine intestinal models that recapitulate the multicellular complexity of the gastrointestinal tract. Here, we generated swine enteroids from the intestinal crypt stem cells of the duodenum, jejunum, or ileum and found that the generated enteroids are able to satisfactorily recapitulate the complicated intestinal epitheliumin vivoand are susceptible to infection by PEDV. PEDV infected multiple types of cells, including enterocytes, stem cells, and goblet cells, and exhibited segmental infection discrepancies compared with ileal enteroids and colonoids, and this finding was verifiedin vivo. Moreover, the clinical isolate PEDV-JMS propagated better in ileal enteroids than the cell-adapted isolate PEDV-CV777, and PEDV infection suppressed interferon (IFN) production early during the infection course. IFN lambda elicited a potent antiviral response and inhibited PEDV in enteroids more efficiently than IFN alpha (IFN-α). Therefore, swine enteroids provide a novelin vitromodel for exploring the pathogenesis of PEDV and for thein vitrostudy of the interplay between a host and a variety of swine enteric viruses.IMPORTANCEPEDV is a highly contagious enteric coronavirus that causes significant economic losses, and the lack of a goodin vitromodel system is a major roadblock to an in-depth understanding of PEDV pathogenesis. Here, we generated a porcine intestinal enteroid model for PEDV infection. Utilizing porcine intestinal enteroids, we demonstrated that PEDV infects multiple lineages of the intestinal epithelium and preferably infects ileal enteroids over colonoids and that enteroids prefer to respond to IFN lambda 1 over IFN-α. These events recapitulate the events that occurin vivo. This study constitutes the first use of a primary intestinal enteroid model to investigate the susceptibility of porcine enteroids to PEDV and to determine the antiviral response following infection. Our study provides important insights into the events associated with PEDV infection of the porcine intestine and provides a valuablein vitromodel for studying not only PEDV but also other swine enteric viruses.

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Tissue underlying the intestinal epithelium elicits proliferation of intestinal stem cells following cytotoxic damage

Cell and Tissue Research ◽

10.1007/s00441-015-2111-1 ◽

2015 ◽

Vol 361 (2) ◽

pp. 427-438 ◽

Cited By ~ 15

Author(s):

Kristen M Seiler ◽

Erica L Schenhals ◽

Richard J von Furstenberg ◽

Bhavya K Allena ◽

Brian J Smith ◽

...

Keyword(s):

Stem Cells ◽

Intestinal Epithelium ◽

Intestinal Stem Cells

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Percoll Gradient Separation of Cord Blood Mononuclear Cells Reveals the Presence of a Novel Population of CXCR4+ Oct-4+ Small Embryonic-Like Stem Cells.

Blood ◽

10.1182/blood.v106.11.1069.1069 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1069-1069 ◽

Cited By ~ 1

Author(s):

Magda Kucia ◽

Krzysztof Oldak ◽

Mariusz Z. Ratajczak ◽

Janina Ratajczak ◽

Zygmunt Pojda

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cord Blood ◽

Intestinal Epithelium ◽

Skeletal Muscles ◽

Mononuclear Cells ◽

Neural Tissue ◽

Open Chromatin ◽

Percoll Gradient ◽

Cell Type

Abstract Cord blood (CB) mononuclear cells (MNC) were demonstrated to contribute to organ/tissue regeneration, however, the identity of the specific cell type(s) involved remains unknown. Recently, it had been identified in adult bone marrow (BM) a mobile, SDF-1-, HGF/SF- and LIF-responsive population of CXCR4+ non-hematopoietic MNC that expresses markers (RQ-PCR, immunhistochemistry) of early pluripotent/tissue committed stem cells (TCSC) for skeletal muscles, heart, neural tissue, liver, pancreas, epidermis and intestinal epithelium (Leukemia2004:18;29–40). We hypothesized that a similar population of these rare cells may also be present in CB, however, their final yield may depend on the method of MNC preparation. We hypothesized that since these cells are very small (~3–5 μm), they may cross a Ficoll-Paque gradient or sedimentate more rapidly than other CB MNC and as result of this are lost during routine CB preparations. Thus, taking in consideration their small size, in order to evaluate if these cells are present in CB, we isolated small CB MNC by employing Percoll gradient (1.078–1.095). We found that this allows us to isolate from CB a population of small cells (2.5% of the total number of MNC) that is enriched in a population of TCSC/PSC(~0.002% of MNC) that we have originally identified in BM. Accordingly, these CB-derived TCSC/PSC CXCR4+ cells are very small (~3μm), posses large nuclei that contain embryonic stem cell type open chromatin (euchromatin), and express several markers for skeletal muscles, heart, neural tissue, liver, pancreas, epidermis and intestinal epithelium as well as pluripotent stem cell (PSC) transcription factors such as Oct-4, Nanog and Rex-1. In vitro cultures of CB-derived small TCSC/PSC are able to grow neurospheres that gave rise to neuronal (β-III tubulin+, nestin+) and macroglia (O4+, MBP+, GFAP+) lineages and cardiomyocytes (β-myosin heavy chain+, α-sarcomeric actin+). Based on this we conclude that CB contains embryonic-like stem cells which may be lost during routine procedures to isolate MNC. Thus, Percoll gradient centrifugation allows for optimal isolation of these small CXCR4+ PSC/TCSC and we postulate that the CB tissue/organ regenerating potential may be much higher than initially postulated and we are currently testing this hypothesis in vivo in animal models.

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Delayed activation of the DNA replication licensing system in Lgr5(+) intestinal stem cells

10.1101/177477 ◽

2017 ◽

Author(s):

T.D. Carroll ◽

I.P. Newton ◽

Y. Chen ◽

J.J. Blow ◽

I. Näthke

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Dna Replication ◽

Intestinal Epithelium ◽

Intestinal Stem Cells ◽

Temporal Window ◽

Restriction Point ◽

Interphase Cells ◽

Small Intestinal ◽

Functional Restriction

ABSTRACTDuring late mitosis and early G1, replication origins are licensed for replication by binding to double hexamers of MCM2-7. Here, we investigate how licensing and proliferative commitment are coupled in the small-intestinal epithelium. We developed a method for identifying cells in intact tissue containing DNA-bound MCM2-7. Interphase cells above the transit-amplifying compartment had no DNA-bound MCM2-7, but still expressed MCM2-7 protein, suggesting that licensing is inhibited immediately upon differentiation. Strikingly, we found most proliferative Lgr5(+) stem cells are in an unlicensed state. This suggests that the elongated cell-cycle of intestinal stem-cells is caused by an increased G1 length, characterised by dormant periods with unlicensed origins. Significantly, the unlicensed state is lost In Apc mutant epithelium, which lacks a functional restriction point, causing licensing immediately upon G1 entry. We propose that the unlicensed G1 of intestinal stem cells creates a temporal window when proliferative fate decisions can be made.

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