NFAT5 dictates crosstalk between intestinal epithelial regenerative capacity and microbiota in murine colitis models
Se-Hyeon Park, Dae Hee Cheon, Yu-Mi Kim, Yeji Choi, Yong-Joon Cho, Bong-Ki Hong, Sang-Hyun Cho, Mi-Na Kweon, Hyug Moo Kwon, Eugene B. Chang, Donghyun Kim, Wan-Uk Kim
Se-Hyeon Park, Dae Hee Cheon, Yu-Mi Kim, Yeji Choi, Yong-Joon Cho, Bong-Ki Hong, Sang-Hyun Cho, Mi-Na Kweon, Hyug Moo Kwon, Eugene B. Chang, Donghyun Kim, Wan-Uk Kim
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Research Article Gastroenterology Immunology

NFAT5 dictates crosstalk between intestinal epithelial regenerative capacity and microbiota in murine colitis models

Abstract

Hypertonic and hyperosmolar stimuli frequently pose challenges to the intestinal tract. Therefore, a resilient epithelial barrier is essential for maintaining gut homeostasis in the presence of osmotic perturbations. Nuclear factor of activated T cells 5 (NFAT5), an osmosensitive transcription factor, primarily maintains cellular homeostasis under hypertonic conditions. However, the osmoprotective role of NFAT5 in enterocyte homeostasis is poorly understood. Here, we demonstrate that NFAT5 was critical for the survival and proliferation of intestinal epithelial cells (IECs) and that its deficiency accelerated chemically induced or spontaneous colitis in mice. Mechanistically, NFAT5 promoted the survival of IECs and the renewal of intestinal stem cells, thereby regulating the production of mucus and antimicrobial compounds, including RegIII and lysozyme, which consequently shape the gut microbial composition to prevent colitis. Transcriptome analysis identified HSP70 as a key downstream target of NFAT5 in epithelial regeneration. Loss- and gain-of-function experiments involving HSP70 revealed that NFAT5 mitigated experimental colitis through IEC Hsp70, which protected stem cells from inflammation-induced injury and maintained barrier function. In conclusion, our study demonstrates what we believe to be a previously unknown role for NFAT5 in dictating the crosstalk between intestinal stem cells and the microbiota, underscoring the importance of the NFAT5/HSP70 axis in maintaining epithelial regeneration related to gut barrier function, balancing microbial composition, and subsequently preventing colitis progression.

Authors

Se-Hyeon Park, Dae Hee Cheon, Yu-Mi Kim, Yeji Choi, Yong-Joon Cho, Bong-Ki Hong, Sang-Hyun Cho, Mi-Na Kweon, Hyug Moo Kwon, Eugene B. Chang, Donghyun Kim, Wan-Uk Kim

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Figure 10

NFAT5 deficiency exacerbates spontaneous colitis in Il10-deficient mice by influencing gut microbiota composition and impairing epithelial regenerative capacity.

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NFAT5 deficiency exacerbates spontaneous colitis in Il10-deficient mice ...
(A) Relative expression levels of Hspa1b mRNA in L-IECs and S-IECs from separately housed 10-week-old Il10–/– Nfat5+/+ and Il10–/– Nfat5+/– mice were quantified by qRT-PCR and normalized to Gapdh mRNA. (B–E) IHC staining for HSP70 (B) and mucin 2 (C) in colonic tissues, as well as mucin 2 (D) and lysozyme (E) in ileal tissues, was carried out for separately housed Il10–/– Nfat5+/+ and Il10–/– Nfat5+/– mice at 10 weeks of age. Representative images are and the corresponding graphs are shown. (F) Organoids were generated by culturing intestinal crypts isolated from separately housed Il10–/– Nfat5+/+ and Il10–/– Nfat5+/– mice in Matrigel for 5 days. Crypts were isolated from the small intestine of individual mice, and the efficiency of organoid formation was assessed by measuring both the number and surface area of organoids generated per well. Multiple images were acquired from each well, and the surface area of all organoids within these images was quantified. Representative images of the formed organoids are shown. Each dot represents an individual mouse, with group means indicated by horizontal lines (A–E). Data shown in A–E are representative of at least 2 independent experiments; data in F are representative of 3 independent experiments and are presented as box-and-whiskers plots (minimum-to-maximum, line at median). *P < 0.05 and **P < 0.01, and ***P < 0.001 by Mann-Whitney U test (A–E) and unpaired, 2-tailed Student’s t test (F). Scale bars: 100 μm and 1,000 μm (C, F, and I).