2. Academic Validation
  3. Paneth cell-derived iNOS is required to maintain homeostasis in the intestinal stem cell niche

Paneth cell-derived iNOS is required to maintain homeostasis in the intestinal stem cell niche

  • J Transl Med. 2023 Nov 25;21(1):852. doi: 10.1186/s12967-023-04744-w.
Lingxiao Huang # 1,  Zhenni Xu # 1,  Xudan Lei 1,  Yujun Huang 1 2,  Siyu Tu 1 2,  Lu Xu 1,  Jieying Xia 3,  Dengqun Liu 4
Affiliations

Affiliations

  • 1 Radiation Oncology Key Laboratory of Sichuan Province, Department of Experimental Research, Sichuan Cancer Hospital & Institute, Sichuan Clinical Research Center for Cancer, Sichuan Cancer Center, Affiliated Cancer Hospital of University of Electronic Science and Technology of China, Chengdu, 610041, China.
  • 2 School of Basic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China.
  • 3 Animal Experiment Center of Sichuan Academy of Traditional Chinese Medicine Sciences, Chengdu, 610041, China.
  • 4 Radiation Oncology Key Laboratory of Sichuan Province, Department of Experimental Research, Sichuan Cancer Hospital & Institute, Sichuan Clinical Research Center for Cancer, Sichuan Cancer Center, Affiliated Cancer Hospital of University of Electronic Science and Technology of China, Chengdu, 610041, China. [email protected].
  • # Contributed equally.
PMID: 38007452 DOI: 10.1186/s12967-023-04744-w
Abstract

Background: Mammalian intestinal epithelium constantly undergoes rapid self-renewal and regeneration sustained by intestinal stem cells (ISCs) within crypts. Inducible nitric oxide synthase (iNOS) is an important regulator in tissue homeostasis and inflammation. However, the functions of iNOS on ISCs have not been clarified. Here, we aimed to investigate the expression pattern of inducible nitric oxide synthase (iNOS) within crypts and explore its function in the homeostatic maintenance of the ISC niche.

Methods: Expression of iNOS was determined by tissue staining and qPCR. iNOS-/- and Lgr5 transgenic mice were used to explore the influence of iNOS ablation on ISC proliferation and differentiation. Enteroids were cultured to study the effect of iNOS on ISCs in vitro. Ileum samples from wild-type and iNOS-/- mice were collected for RNA-Seq to explore the molecular mechanisms by which iNOS regulates ISCs.

Results: iNOS was physiologically expressed in Paneth cells. Knockout of iNOS led to apparent morphological changes in the intestine, including a decrease in the small intestine length and in the heights of both villi and crypts. Knockout of iNOS decreased the number of Ki67+ or BrdU+ proliferative cells in crypts. Loss of iNOS increased the number of Olfm4+ ISCs but inhibited the differentiation and migration of Lgr5+ ISCs in vivo. iNOS depletion also inhibited enteroid formation and the budding efficiency of crypts in vitro. Moreover, iNOS deficiency altered gluconeogenesis and the adaptive immune response in the ileum transcriptome.

Conclusion: Paneth cell-derived iNOS is required to maintain a healthy ISC niche, and Knockout of iNOS hinders ISC function in mice. Therefore, iNOS represents a potential target for the development of new drugs and other therapeutic interventions for intestinal disorders.

Keywords

Differentiation; Intestinal stem cell; Paneth cell; Proliferation; iNOS.

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