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Krüppel-like Factor 5 Regulates Stemness, Lineage Specification, and Regeneration of Intestinal Epithelial Stem Cells

Chang-Kyung Kim1, Madhurima Saxena2, Kasmika Maharjan1

  • 1Department of Medicine, Stony Brook University Renaissance School of Medicine, Stony Brook, New York.

Cellular and Molecular Gastroenterology and Hepatology
|November 29, 2019
PubMed
Abstract

Insights

Krüppel-like factor 5 (KLF5) is essential for intestinal stem cell (ISC) self-renewal and regeneration. Its absence leads to faster proliferation but impaired self-renewal and differentiation, impacting epithelial repair.

Area of Science:

  • Stem cell biology
  • Gastrointestinal biology
  • Molecular genetics

Background:

  • Intestinal stem cells (ISCs) possess self-renewal and differentiation capabilities crucial for gut homeostasis.
  • WNT and NOTCH signaling pathways are known regulators of ISC function, but underlying molecular mechanisms remain incompletely understood.
  • Krüppel-like factor 5 (KLF5) role in ISC regulation was investigated.

Purpose of the Study:

  • To elucidate the function of Krüppel-like factor 5 (KLF5) in maintaining intestinal stem cell (ISC) identity and function.
  • To understand the molecular mechanisms by which KLF5 regulates ISC self-renewal and differentiation.
  • To assess the impact of KLF5 loss on intestinal epithelial regeneration after injury.

Main Methods:

  • Utilized genetically engineered mouse models (Lgr5ΔKlf5 and Lgr5Ctrl) with tamoxifen-inducible Klf5 deletion.
  • Administered tamoxifen to induce Cre recombinase activity and delete Klf5 in Lgr5+ ISCs.
  • Performed whole-body irradiation (TBI) to induce intestinal injury.
  • Conducted immunofluorescence (IF) analysis and next-generation sequencing on isolated intestinal tissues and organoids.
  • Employed fluorescence-activated cell sorting (FACS) for single-cell isolation.

Main Results:

  • Klf5-deficient ISCs exhibited accelerated proliferation but failed self-renewal, leading to ISC compartment depletion.
  • Transcriptome analysis indicated loss of ISC identity and premature differentiation in Klf5-null ISCs.
  • Following irradiation, Klf5-null progenitor cells were unable to dedifferentiate and regenerate the intestinal epithelium.
  • KLF5 absence inactivated key enhancer elements and WNT/NOTCH target genes.
  • Human intestinal tissues showed elevated KLF5 levels in regenerating epithelia compared to healthy controls.

Conclusions:

  • Klf5 is indispensable for ISC self-renewal, lineage specification, and progenitor cell dedifferentiation.
  • KLF5 regulates ISC functions by controlling epigenetic and transcriptional activities of stem cell-specific genes.
  • Targeting KLF5 presents a potential strategy for modulating intestinal stem cell functions.
Keywords:
Epigenetic RegulationIntestinal Stem CellMultipotent DifferentiationTissue Regeneration

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