Dataset Information

Analysis by RNA-seq of the transcriptional profile of three sub-populations of mouse colon epithelial cells (Lgr5+ columnar basal cells, goblet cells, enterocytes).

ABSTRACT: In this dataset, we report the analysis by RNA-sequencing (RNA-seq) of the transcriptional profile of three sub-populations of epithelial cells contained in the mouse colonic epithelium: 1) Epcam+/Cd44+/Kit-neg cells, which correspond to a population of epithelial cells located at the bottom of colonic crypts and enriched in Lgr5+ columnar basal cells (CBCs); 2) Epcam+/Cd44+/Kit+ cells, which correspond to a population of epithelial cells located at the bottom of colonic crypts and enriched in goblet cells; 3) Epcam+/Cd44-neg/Cd66-high cells, which correspond to a population of epithelial cells located at the top of colonic crypts and enriched in mature enterocytes (Rothenberg et al., Gastroenterology, 142:1195-1205, 2012).

ORGANISM(S): Mus musculus

PROVIDER: GSE202483 | GEO | 2024/05/01

REPOSITORIES: GEO

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Project description:In human colon cancer, the malignant component of tumor tissues often contains multiple sub-types of cancer cells, whose transcriptional profiles and surface marker phenotypes correspond to those of the epithelial lineages that are found in normal colonic crypts (e.g., goblet cells, enterocytes, LGR5+ columnar basal cells). Among those various sub-types of malignant cells, those characterized by a surface marker phenotype that is characteristic of epithelial stem/progenitor cells residing at the bottom of colonic crypts (EpCAM+, CD44+, CD166+) are enriched in cells with "cancer stem cell" (CSC) properties, such as self-renewal (i.e., the capacity to sustain the formation of new tumors upon serial xeno-transplantation in immune-deficient animals) and multi-lineage differentiation (i.e., the capacity to sustain the formation of other cell-types, thus reconstituting the heterogeneous population of the parent tumors from which they have been isolated). Cell populations with "cancer stem cell" (CSC) properties are known to be preferentially resistant to several cytotoxic agents used in conventional chemotherapy, but the molecular mechanisms underpinning this property remain poorly understood. In this dataset, we report the analysis by gene-expression microarrays of the transcriptional profile of two sub-populations of human colon cancer cells: 1) cells with a "bottom-of-the-crypt" phenotype (EpCAM+, CD44+, CD166+), which are known to be enriched in "cancer stem cells" (CSCs); and 2) cells with a "top-of-the-crypt" phenotype (EpCAM+, CD44neg, CD166neg), which are known to be non-tumorigenic upon xeno-transplantation in immune-deficient mice. The two populations were purified in parallel by fluorescence activated cell sorting (FACS), starting from solid tumors established by sub-cutaneous (s.c.) engraftment in immune-deficient mice of a patient-derived xenograft (PDX) line representative of a moderately differentiated (G2) primary colon carcinoma. To identify genes whose expression is modulated by in vivo exposure to chemotherapy (and thus potentially involved in the mechanistic regulation of chemo-resistance), we compared the transcriptional profile of cells purified from tumors that were exposed to 4 weeks of in vivo treatment with either irinotecan (CPT-11) or a placebo control (saline solution).

Project description:There is a gradient of M-NM-2-catenin expression along the colonic crypt axis with the highest levels at the crypt bottom. However, it remains unclear whether different levels of canonical Wnt signaling exert distinct roles in the colonic epithelium. In the present study, we first showed that the canonical Wnt signaling is active in the proliferative compartment of normal colonic crypts by separating actively proliferating progenitor cells from non-proliferating cells in the colon using transgenic mice expressing a histone H2B-green fluorescent protein (GFP) fusion protein under the control of a tetracycline responsive regulatory element. Subsequently, we investigated the dose-dependent effect of canonical Wnt activation on colonic epithelial differentiation by controlling the expression levels of stabilized M-NM-2-catenin using a doxycycline-inducible transgenic system in mice. We show that elevated levels of Wnt signaling induce the amplification of Lgr5+ cells, which is accompanied by crypt fission and a reduction in cell proliferation among progenitor cells. In contrast, lower levels of M-NM-2-catenin induction enhanced cell proliferation rates of epithelial progenitors without affecting crypt fission rates. Notably, slow-cycling cells produced by M-NM-2-catenin activation exhibit activation of Notch signaling and the treatment of M-NM-2-catenin expressing mice with a Notch inhibitor turned such slow-cycling cells into actively proliferating cells. Our results indicate that the activation of the canonical Wnt signaling pathway is sufficient for de novo crypt formation, and suggest that different levels of canonical Wnt activations, in cooperation with Notch signaling, establish a hierarchy of slower-cycling stem cells and faster-cycling progenitor cells characteristic for the colonic epithelium. To separate actively proliferating cells from non-proliferating cells in normal colon, we used transgenic mice expressing a histone H2B-GFP fusion protein under the control of a tetracycline responsive regulatory element (H2B-GFP mice). In H2B-GFP mice, all colonic crypt cells exhibited strong nuclear GFP signal 7 days after doxycycline administration. When doxycycline was withdrawn for 2 days after the labeling period, nuclear GFP signal was diluted in proliferating cells whereas non-proliferating cells retained GFP. GFPhigh non-proliferating and GFPlow proliferating epithelial cells were sorted from the isolated crypts by FACS for microarray analysis. In addition, to investigate the effects of acute Wnt activation in colon, we generated doxycycline-inducible M-NM-2-catenin mice. Colonic crypts were isolated from untreated mice and mice fed doxycycline in the drinking water (2.0 mg/ml) for 5 days and subjected to microarray analysis.

Dataset Information

Analysis by RNA-seq of the transcriptional profile of three sub-populations of mouse colon epithelial cells (Lgr5+ columnar basal cells, goblet cells, enterocytes).

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