Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium

Mustata, Roxana C.; Vasile, Gabriela; Vallone, Valeria Fernández; Strollo, Sandra; Lefort, Anne; Libert, Frédérick; Monteyne, Daniel; Pérez‐Morga, David; Vassart, Gilbert; Garcia, M.

doi:10.1016/j.celrep.2013.09.005

Cited by 238 publications

(321 citation statements)

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“…Importantly, loss of Tet1 also impairs budding organoid-forming capacity and thus de novo formation or expansion of stem cells in crypt culture. It was reported previously (Mustata et al 2013) that when organoids are grown from the fetal intestine, they form spheroids instead of budding organoids and exhibit low levels of Wnt target gene expression. The organoids that we obtained from the Tet1-deleted postnatal intestine resemble these spheroids, suggesting an underdeveloped state.…”

Section: Discussionmentioning

confidence: 99%

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Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

et al. 2016

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Methylated cytosines are associated with gene silencing. The ten-eleven translocation (TET) hydroxylases, which oxidize methylated cytosines to 5-hydroxymethylcytosine (5hmC), are essential for cytosine demethylation. Gene silencing and activation are critical for intestinal stem cell (ISC) maintenance and differentiation, but the potential role of TET hydroxylases in these processes has not yet been examined. Here, we generated genome-wide maps of the 5hmC mark in ISCs and their differentiated progeny. Genes with high levels of hydroxymethylation in ISCs are strongly associated with Wnt signaling and developmental processes. We found Tet1 to be the most abundantly expressed Tet gene in ISCs; therefore, we analyzed intestinal development in Tet1-deficient mice and determined that these mice are growth-retarded, exhibit partial postnatal lethality, and have significantly reduced numbers of proliferative cells in the intestinal epithelium. In addition, the Tet1-deficient intestine displays reduced organoidforming capacity. In the Tet1-deficient crypt, decreased expression of Wnt target genes such as Axin2 and Lgr5 correlates with lower 5hmC levels at their promoters. These data demonstrate that Tet1-mediated DNA hydroxymethylation plays a critical role in the epigenetic regulation of the Wnt pathway in intestinal stem and progenitor cells and consequently in the self-renewal of the intestinal epithelium.

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Section: Discussionmentioning

confidence: 99%

“…7A-C). This suggests that the state of postnatal crypts from Tet1 −/− mice is closer to fetal than adult crypts (Fordham et al 2013;Mustata et al 2013).…”

Section: Tet1 Is Required For Postnatal Intestinal Developmentmentioning

confidence: 95%

Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

et al. 2016

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“…Several recently published studies analyze the differences between fetal progenitors and adult ISCs (Fordham et al, 2013;Mustata et al, 2013). Fordham and colleagues report that intestinal progenitor cell maturation correlates with increased Wnt signaling.…”

Section: Discussionmentioning

confidence: 99%

Dnmt1 is essential to maintain progenitors in the perinatal intestinal epithelium

et al. 2015

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The DNA methyltransferase Dnmt1 maintains DNA methylation patterns and genomic stability in several in vitro cell systems. Ablation of Dnmt1 in mouse embryos causes death at the postgastrulation stage; however, the functions of Dnmt1 and DNA methylation in organogenesis remain unclear. Here, we report that Dnmt1 is crucial during perinatal intestinal development. Loss of Dnmt1 in intervillus progenitor cells causes global hypomethylation, DNA damage, premature differentiation, apoptosis and, consequently, loss of nascent villi. We further confirm the crucial role of Dnmt1 during crypt development using the in vitro organoid culture system, and illustrate a clear differential requirement for Dnmt1 in immature versus mature organoids. These results demonstrate an essential role for Dnmt1 in maintaining genomic stability during intestinal development and the establishment of intestinal crypts.

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“…Crypts isolated from fetal mouse intestine form spontaneously spheroids, a potential that is rapidly lost after birth (29), suggesting that postnatal IESCs become dependent on external factors. Accordingly, crypts mature between embryonic day (E) 16.5 and the first weeks after birth (30).…”

Section: Cd34mentioning

confidence: 99%

CD34 ⁺ mesenchymal cells are a major component of the intestinal stem cells niche at homeostasis and after injury

Stzepourginski

Nigro

Jacob

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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The intestinal epithelium is continuously renewed by intestinal epithelial stem cells (IESCs) positioned at the base of each crypt. Mesenchymal-derived factors are essential to maintain IESCs; however, the cellular composition and development of such mesenchymal niche remains unclear. Here, we identify pericryptal CD34 + Gp38 + αSMA -mesenchymal cells closely associated with Lgr5+ IESCs. We demonstrate that CD34 + Gp38 + cells are the major intestinal producers of the niche factors Wnt2b, Gremlin1, and R-spondin1, and are sufficient to promote maintenance of Lgr5 + IESCs in intestinal organoids, an effect mainly mediated by Gremlin1. CD34+ Gp38 + cells develop after birth in the intestinal submucosa and expand around the crypts during the third week of life in mice, independently of the microbiota. We further show that pericryptal CD34+ cells are rapidly activated by intestinal injury, up-regulating niche factors Gremlin1 and R-spondin1 as well as chemokines, proinflammatory cytokines, and growth factors with key roles in gut immunity and tissue repair, including IL-7, Ccl2, Ptgs2, and Amphiregulin. Our results indicate that CD34 + Gp38 + mesenchymal cells are programmed to develop in the intestine after birth to constitute a specialized microenvironment that maintains IESCs at homeostasis and contribute to intestinal inflammation and repair after injury.T he adult intestinal epithelium is one of the most rapidly selfrenewing tissues in mammals. Intestinal epithelial cells renewal is ensured by intestinal epithelial stem cells (IESCs) located in the crypts and identified by expression of Lgr5 (1). IESCs are responsible for the continuous production of rapidly dividing transit-amplifying (TA) cells and of Paneth cells while maintaining the size of their own population. Upon leaving the crypts, TA cells proliferate and migrate upwards, differentiating into enterocytes, goblet cells, Tuft cells, and enteroendocrine cells, before undergoing apoptosis at the villus tip and being shed into the intestinal lumen (2).A complex gradient of factors maintains IESC stemness and proliferation, and supports enterocyte differentiation along the crypt-villus axis. Notably, Wnt signals are required to maintain the IESC niche (3, 4). In the crypts, Paneth cells express factors that promote stem cells growth, including the Wnt ligand Wnt3a, Notch ligands (Dll4, Dll1), and epidermal growth factor (EGF) (3,5, 6). In addition, a number of factors produced by mesenchymal cells have an essential role in the maintenance of IESCs such as Wnt2b, a canonical Wnt ligand that activates the Wnt/ β-catenin pathway (7); the Lgr4/5 ligand R-spondin1 (Rspo1), a strong mitogen for Wnt-responsive intestinal crypts (8-10); and Gremlin1 (Grem1), a bone morphogenetic protein (BMP) antagonist (11, 12). As epithelial cells ascend the crypt, BMPs produced by alpha-smooth muscle actin positive (αSMA + ) myofibroblasts restrain Wnt-induced epithelial proliferation while promoting their differentiation into secretory or absorptive epithelial cells (13,14). W...

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Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium

Cited by 238 publications

References 50 publications

Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

Dnmt1 is essential to maintain progenitors in the perinatal intestinal epithelium

CD34 ⁺ mesenchymal cells are a major component of the intestinal stem cells niche at homeostasis and after injury

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Identification of Lgr5-Independent Spheroid-Generating Progenitors of the Mouse Fetal Intestinal Epithelium

Cited by 238 publications

References 50 publications

Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

Epigenetic regulation of intestinal stem cells by Tet1-mediated DNA hydroxymethylation

Dnmt1 is essential to maintain progenitors in the perinatal intestinal epithelium

CD34 + mesenchymal cells are a major component of the intestinal stem cells niche at homeostasis and after injury

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CD34 ⁺ mesenchymal cells are a major component of the intestinal stem cells niche at homeostasis and after injury