KLF5 Governs Stemness in the Adult Intestinal Stem Cell Niche
| Autor: | Sayantani Goswami, Nan Gao |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Male
TF transcription factor Epigenesis Genetic Receptors G-Protein-Coupled Mice RFP red fluorescent protein GSEA gene set enrichment analysis IRR irradiation RNA-Seq Cell Self Renewal Intestinal Mucosa Stem Cell Niche Wnt Signaling Pathway Cells Cultured Original Research Stem Cells ASCL2 achaete-scute family bHLH transcription factor 2 EdU 5-ethynyl-2′-deoxyuridine Gastroenterology Colitis EGFP enhanced green fluorescent protein Enteritis Stem cell niche ChIP-seq chromatin immunoprecipitation assay with sequencing Cell biology Organoids Adult Stem Cells KLF5 Krüppel-like factor 5 Editorial ISC intestinal stem cell Female Colorectal Neoplasms Epigenetic Regulation Whole-Body Irradiation Transcriptional Activation inorganic chemicals Primary Cell Culture Kruppel-Like Transcription Factors Mice Transgenic LGR5 leucine rich repeat containing G protein-coupled receptor 5 Multipotent Differentiation IGV Integrative Genomics Viewer Tissue Regeneration Biology digestive system Intestinal Stem Cell Animals Humans Regeneration Cell Lineage lcsh:RC799-869 Radiation Injuries Hepatology RT-qPCR reverse transcriptase quantitative polymerase chain reaction fungi RNA-seq RNA sequencing TA transit amplifying Disease Models Animal Case-Control Studies H&E hematoxylin and eosin TSS transcription start site lcsh:Diseases of the digestive system. Gastroenterology TUNEL terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling Transcription Factors |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology Cellular and Molecular Gastroenterology and Hepatology, Vol 9, Iss 4, Pp 705-706 (2020) |
| ISSN: | 2352-345X |
| DOI: | 10.1016/j.jcmgh.2019.12.007 |
| Popis: | Background & Aims Self-renewal and multipotent differentiation are cardinal properties of intestinal stem cells (ISCs), mediated in part by WNT and NOTCH signaling. Although these pathways are well characterized, the molecular mechanisms that control the ‘stemness’ of ISCs are still not well defined. Here, we investigated the role of Krüppel-like factor 5 (KLF5) in regulating ISC functions. Methods We performed studies in adult Lgr5EGFP-IRES-creERT2;Rosa26LSLtdTomato (Lgr5Ctrl) and Lgr5EGFP-IRES-creERT2;Klf5fl/fl;Rosa26LSLtdTomato (Lgr5ΔKlf5) mice. Mice were injected with tamoxifen to activate Cre recombinase, which deletes Klf5 from the intestinal epithelium in Lgr5ΔKlf5 but not Lgr5Crtl mice. In experiments involving irradiation, mice were subjected to 12 Gy total body irradiation (TBI). Tissues were collected for immunofluorescence (IF) analysis and next generation sequencing. Oganoids were derived from fluoresecence activated cell sorted- (FACS-) single cells from tamoxifen-treated Lgr5ΔKlf5 or Lgr5Crtl mice and examined by immunofluorescence stain. Results Lgr5+ ISCs lacking KLF5 proliferate faster than control ISCs but fail to self-renew, resulting in a depleted ISC compartment. Transcriptome analysis revealed that Klf5-null Lgr5+ cells lose ISC identity and prematurely differentiate. Following irradiation injury, which depletes Lgr5+ ISCs, reserve Klf5-null progenitor cells fail to dedifferentiate and regenerate the epithelium. Absence of KLF5 inactivates numerous selected enhancer elements and direct transcriptional targets including canonical WNT- and NOTCH-responsive genes. Analysis of human intestinal tissues showed increased levels of KLF5 in the regenerating epithelium as compared to those of healthy controls. Conclusion We conclude that ISC self-renewal, lineage specification, and precursor dedifferentiation require KLF5, by its ability to regulate epigenetic and transcriptional activities of ISC-specific gene sets. These findings have the potential for modulating ISC functions by targeting KLF5 in the intestinal epithelium. Graphical abstract |
| Databáze: | OpenAIRE |
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