Dataset Information

YTHDF2-KIF26B-Wnt signaling forms a positive-feedback regulatory loop to maintain ISC stemness

ABSTRACT: m6A modification plays a regulatory role in the stemness of small intestinal epithelial stem cells (ISCs), which is crucial for the renewal of intestinal epithelium and the maintenance of homeostasis. Although existing research has demonstrated that METTL3, as an m6A ‘writer’, is a crucial protein in the regulation of the stemness of ISCs, the mechanism by which the function of downstream ‘reader’ proteins is regulated has not been fully elucidated. In this study, we found that the knockout of Ythdf2 could significantly down-regulate the expression of the stemness marker gene Lgr5, suggesting that this ‘reader’ protein plays an important role in the regulation of the stemness of ISCs. Upon exploring the specific downstream mechanisms, we discovered that YTHDF2 promoted the Wnt signaling pathway by regulating the expression of KIF26B. This helps maintain the expression of Lgr5 in stem cells, thus preserving the stemness of these cells. Meanwhile, the expression of YTHDF2 itself is also regulated by the Wnt signaling pathway. Overall, YTHDF2 forms a positive-feedback regulatory loop centered around the Wnt signaling pathway. This loop ensures that ISCs remain in a stem-cell state, enabling the intestinal epithelium to function stably and properly.

ORGANISM(S): Mus musculus

PROVIDER: GSE290302 | GEO | 2025/05/12

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Background & Aims: Hierarchical organization of intestine relies on their stem cells by self-renew and producing committed progenitors. Although signals like Wnt are known to animate the continued renewal by maintaining intestinal stem cells (ISCs) activity, molecular mechanisms especially E3 ubiquitin ligases that modulate ISCs ‘stemness’ and supportive niche have not been well understood. Here, we investigated the role of Cullin 4B (Cul4b) in regulating ISC functions. Methods: We generated mice with intestinal epithelial-specific disruption of Cul4b (pVillin-cre; Cul4bfn/Y), inducible disruption of Cul4b (Lgr5-creERT2; Cul4bfn/Y, CAG-creERT2; Cul4bfn/Y) and their control (Cul4bfn/Y). Intestinal tissues were analyzed by histology, immunofluorescence, RNA sequencing and mass spectrum. Intestinal organoids deprived from mice with pVillin-Cre; Cul4bfn/Y, Lgr5-Cre; Cul4bfn/Y, Tg-Cul4b and their controls were used in assays to measure intestinal self-renewal, proliferation and differentiation. Wnt signaling and intestinal markers were analyzed by immunofluorescence and immunoblot assays. Differential proteins upon Cul4b ablation or Cul4b-interacting proteins were identified by mass spectrometry. Results: Cul4b specifically located at ISCs zone. Block of Cul4b impaired intestinal homeostasis maintenance by reduced self-renewal and proliferation. Transcriptome analysis revealed that Cul4b-null intestine lose ISC characterization and showed disturbed ISC niche. Mechanistically, reactivated Wnt pathway could recover intestinal dysfunction of Cul4b knockout mice. Analysis of differential total and ubiquitylated proteins uncovered the novel targeting substrate of Cullin-Ring ubiquitin ligase 4b (CRL4b), immunity-related GTPase family M member 1 (Irgm1) in intestine. Decreased Irgm1 rescued abnormally interferon signaling, overemphasized autophagy and downstream phosphate proteins in Cul4b knockout mice. Conclusion: We conclude that Cul4b is essential for ISC self-renewal and Paneth cell function by targeting Irgm1 and modulating Wnt signaling. Our results demonstrate that Cul4b is a novel ISC stemness and niche regulator.

Dataset Information

YTHDF2-KIF26B-Wnt signaling forms a positive-feedback regulatory loop to maintain ISC stemness

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets