IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B

IF 7.1 1区医学 Q1 GASTROENTEROLOGY & HEPATOLOGY

Cellular and Molecular Gastroenterology and Hepatology Pub Date : 2024-01-01 DOI:10.1016/j.jcmgh.2023.12.001

Louis R. Parham , Patrick A. Williams , Kay Katada , Shaneice K. Nettleford , Priya Chatterji , Kofi K. Acheampong , Charles H. Danan , Xianghui Ma , Lauren A. Simon , Kaitlyn E. Naughton , Rei Mizuno , Tatiana Karakasheva , Emily A. McMillan , Kelly A. Whelan , Donita C. Brady , Sydney M. Shaffer , Kathryn E. Hamilton

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引用次数: 0

Abstract

Background & Aims

The intestinal epithelium interfaces with a diverse milieu of luminal contents while maintaining robust digestive and barrier functions. Facultative intestinal stem cells are cells that survive tissue injury and divide to re-establish the epithelium. Prior studies have shown autophagic state as functional marker of facultative intestinal stem cells, but regulatory mechanisms are not known. The current study evaluated a post-transcriptional regulation of autophagy as an important factor for facultative stem cell state and tissue regeneration.

Methods

We evaluated stem cell composition, autophagic vesicle content, organoid formation, and in vivo regeneration in mice with intestinal epithelial deletion of the RNA binding protein IGF2 messenger RNA binding protein 1 (IMP1). The contribution of autophagy to resulting in vitro and in vivo phenotypes was evaluated via genetic inactivation of Atg7. Molecular analyses of IMP1 modulation of autophagy at the protein and transcript localization levels were performed using IMP1 mutant studies and single-molecule fluorescent in situ hybridization.

Results

Epithelial Imp1 deletion reduced leucine rich repeat containing G protein coupled receptor 5 cell frequency but enhanced both organoid formation efficiency and in vivo regeneration after irradiation. We confirmed prior studies showing increased autophagy with IMP1 deletion. Deletion of Atg7 reversed the enhanced regeneration observed with Imp1 deletion. IMP1 deletion or mutation of IMP1 phosphorylation sites enhanced expression of essential autophagy protein microtubule-associated protein 1 light chain 3β. Furthermore, immunofluorescence imaging coupled with single-molecule fluorescent in situ hybridization showed IMP1 colocalization with MAP1LC3B transcripts at homeostasis. Stress induction led to decreased colocalization.

Conclusions

Depletion of IMP1 enhances autophagy, which promotes intestinal regeneration via expansion of facultative intestinal stem cells.

Abstract Image

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IGF2BP1/IMP1缺失可通过调节MAP1LC3B增强面干细胞状态

背景& 目的肠道上皮与多种多样的管腔内容物相互作用，同时保持强大的消化和屏障功能。肠道干细胞是在组织损伤后存活下来并分裂以重建上皮的细胞。先前的研究表明，自噬状态是面肠干细胞的功能标志，但其调控机制尚不清楚。本研究评估了转录后对自噬的调控，认为自噬是面性干细胞状态和组织再生的一个重要因素。通过Atg7的遗传失活评估了自噬对体外和体内表型的贡献。利用 IMP1 突变体研究和单分子荧光原位杂交 (smFISH) 对 IMP1 在蛋白质和转录本定位水平上对自噬的调节作用进行了分子分析。我们证实了之前的研究表明，IMP1缺失会增加自噬。Atg7的缺失逆转了Imp1缺失所观察到的再生能力增强。IMP1缺失或IMP1磷酸化位点突变增强了必需的自噬蛋白微管相关蛋白1轻链3β（MAP1LC3B）的表达。此外，免疫荧光成像和 smFISH 显示了 IMP1 与 MAP1LC3B 转录本在平衡状态下的共定位。结论消耗 IMP1 可增强自噬，从而通过扩增肠道干细胞促进肠道再生。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cellular and Molecular Gastroenterology and Hepatology Medicine-Gastroenterology

CiteScore

13.00

自引率

2.80%

发文量

246

审稿时长

42 days

期刊介绍： "Cell and Molecular Gastroenterology and Hepatology (CMGH)" is a journal dedicated to advancing the understanding of digestive biology through impactful research that spans the spectrum of normal gastrointestinal, hepatic, and pancreatic functions, as well as their pathologies. The journal's mission is to publish high-quality, hypothesis-driven studies that offer mechanistic novelty and are methodologically robust, covering a wide range of themes in gastroenterology, hepatology, and pancreatology. CMGH reports on the latest scientific advances in cell biology, immunology, physiology, microbiology, genetics, and neurobiology related to gastrointestinal, hepatobiliary, and pancreatic health and disease. The research published in CMGH is designed to address significant questions in the field, utilizing a variety of experimental approaches, including in vitro models, patient-derived tissues or cells, and animal models. This multifaceted approach enables the journal to contribute to both fundamental discoveries and their translation into clinical applications, ultimately aiming to improve patient care and treatment outcomes in digestive health.