NFAT5 Amplifies Antipathogen Responses by Enhancing Chromatin Accessibility, H3K27 Demethylation, and Transcription Factor Recruitment

Lunazzi, Giulia; Buxadé, María; Riera-Borrull, Marta; Higuera, Laura; Bonnin, Sarah; Encabo, Hector Huerga; Gaggero, Silvia; Reyes-Garau, Diana; Cozzuto, Luca; Ponomarenko, Julia; Aramburu, J.; López-Rodrı́guez, Cristina

doi:10.4049/jimmunol.2000624

Cited by 11 publications

(15 citation statements)

References 64 publications

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“…Mechanically, as a transcription factor, p65 could bind to two binding sites (-591 ~ -582 nt and -299 ~ -290 nt) of IRF-8 gene promotor and increase IRF-8 gene transcription. Here, it is worthy to mention that, since reportedly p65 triggers the transcription of some pro-inflammatory genes such as IL-6, TNF-α and CXCL5 56 , 57 , and serval p65-binding sites are predicted in CCL3 gene promotor, we speculate that p65 could activate CCL3 gene in rat GMCs directly. Thus, we did further experiments, and found that p65 knockdown in the GMCs did not reduce CCL3 gene transcription induced by IRF-8 overexpression, suggesting that p65 does not directly trigger CCL3 gene transcription in the GMCs.…”

Section: Discussionmentioning

confidence: 73%

“…It has been documented that the cells stimulated with external stimulants can rapidly turn on some signal transduction pathways and up-regulate some transcription factors , leading to corresponding target gene expression 56 , 57 . Subsequently, in order to find the upstream regulatory molecules of IRF-8-mediated CCL3/4 production in Thy-1N, we examined the phosphorylation (i.e.…”

Section: Discussionmentioning

confidence: 99%

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Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

Wang¹,

Qian²,

Zhao³

et al. 2022

Int. J. Biol. Sci.

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Mesangioproliferative glomerulonephritis (MsPGN) is a common human kidney disease. Rat Thy-1 nephritis (Thy-1N) is an animal model widely used for the study of MsPGN. Thy-1N is not only sublytic C5b-9-dependent, but also related to pro-inflammatory cytokine production and macrophage (Mφ) accumulation in rat renal tissues. In this study, we found that the expression or phosphorylation of chemokine CCL3/4, CD68 (Mφ marker), IRF-8, PKC-α and NF-κB-p65 (p65) were all up-regulated both in the renal tissues of Thy-1N rats ( in vivo ) and in the glomerular mesangial cells (GMCs) upon sublytic C5b-9 stimulation ( in vitro ). Further experiments in vitro revealed that the phosphorylated PKC-α (p-PKC-α) could promote p65 phosphorylation, and then p-p65 enhanced IRF-8 expression through binding to IRF-8 promotor (-591 ~ -582 nt and -299 ~ -290 nt). Additionally, up-regulation or silencing of IRF-8 gene promoted or reduced CCL3/4 production, and then regulated Mφ chemotaxis. The underlying mechanism involved in IRF-8 binding to CCL3 promoter (-249 ~ -236 nt), which resulted in CCL3 gene transcription. The experiments in vivo showed that knockdown of renal PKC-α, p65, IRF-8 and CCL3/4 genes could inhibit CCL3/4 production, Mφ accumulation, GMC proliferation and proteinuria of Thy-1N rats. Furthermore, p-PKC-α, p-p65, IRF-8, CCL3/4 expression and Mφ accumulation were also increased in the renal tissues of MsPGN patients. Collectively, these findings indicate that sublytic C5b-9 induces CCL3/4 production and Mφ accumulation via PKC-α/p65/IRF-8 axis, and finally aggravates the pathological changes of MsPGN.

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Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

Wang¹,

Qian²,

Zhao³

et al. 2022

Int. J. Biol. Sci.

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“…The cell culture model mimicked the in vivo findings as exposure to hypoxia amplified Pdgfb expression and the release of PDGFB in Nfat5 -/- versus Nfat5 fl/fl MLEC (Figure 4A-C). Although stimulation of gene expression upon knockout of a transcription factor seems counterintuitive, NFAT5 has been reported to alter transcriptional activity by modulating DNA methylation and chromatin accessibility 25 as well as by inhibiting recruitment of transcription factors to enhancer regions of gene promoters 26 . As our results did not substantiate an impact of NFAT5-mediated DNA methylation (Supplement 21) or chromatin accessibility (Supplement 22) on Pdgfb expression under the tested experimental conditions, we investigated whether binding of HIF1α - a transcription factor that is crucial for hypoxia-induced Pdgfb expression in MLEC (Supplement 23) 6 - to the Pdgfb promoter is altered in the presence or absence of NFAT5.…”

Section: Resultsmentioning

confidence: 99%

NFAT5-dependent transcriptional stress control of endothelial cells prevents maladaptive remodeling of pulmonary arterioles in the hypoxic lung

Laban,

Sigmund,

Schlereth

et al. 2023

Preprint

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Objective: Chronic hypoxia causes detrimental structural alterations in the lung, which are partially dependent on stress responses of the endothelium. In this context, we revealed that hypoxia-exposed murine lung endothelial cells (MLEC) activate nuclear factor of activated T-cells 5 (NFAT5/TonEBP) - a transcription factor that adjusts the cellular transcriptome to cope with multiple environmental stressors. Here, we studied the functional relevance of NFAT5 for the control of hypoxia-induced transcription in MLEC. Approach and Results: Targeted ablation of Nfat5 in endothelial cells did not evoke phenotypic abnormalities in normoxia-exposed mice. However, MLEC in Nfat5-deficient mice up-regulated energy- and protein-metabolism-associated gene expression under normobaric hypoxia (10% O2) for seven days as evidenced by microarray- and scRNA-seq-based analyses. Moreover, loss of NFAT5 boosted the expression and release of platelet-derived growth factor B (Pdgfb) - a HIF1a-regulated driver of vascular smooth muscle cell (VSMC) growth - in capillary MLEC of hypoxia-exposed mice, which was accompanied by exaggerated coverage of distal pulmonary arterioles by VSMC, increased pulmonary vascular resistance and impaired right ventricular functions. In vitro, knockout of Nfat5 in cultured MLEC stimulated Pdgfb expression and release after exposure to hypoxia and amplified binding of HIF1a in the Pdgfb promoter region. Conclusion: Collectively, our study identifies NFAT5 as a protective transcription factor required to rapidly adjust the transcriptome of MLEC to hypoxia. Specifically, NFAT5 restricts HIF1a-mediated Pdgfb expression and consequently limits muscularization and resistance of pulmonary arterioles.

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“…qPCR was performed to determine the enrichment of protein-bound DNA. Detailed primer and antibody information can be found in Tables S1 and S2 [45][46][47].…”

Section: Chip-qpcrmentioning

confidence: 99%

N6-methyladenosine of Socs1 modulates macrophage inflammatory response in different stiffness environments

Hu¹,

Li²,

Yuan³

et al. 2022

Int. J. Biol. Sci.

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Macrophages exhibit diverse functions within various tissues during the inflammatory response, and the physical properties of tissues also modulate the characteristics of macrophages. However, the underlying N6-methyladenosine (m 6 A)-associated molecular mechanisms remain unclear. Accordingly, we examined the potential role of m 6 A in macrophage activation and stiffness sensing. Intriguingly, we found that the macrophage inflammatory response and global levels of m 6 A were stiffness-dependent and that this was due to mechanically loosening the chromatin and epigenetic modification (H3K36me2 and HDAC3). In addition, we targeted suppressor of cytokine signalling 1 (Socs1) m 6 A methylation in a stiffness-dependent manner by screening the sequencing data and found that a higher stiffness hydrogel activated Jak-STAT and NFκB signalling and suppressed Fto gene expression. Next, by using the CRISPR/Cas9 system to knockout the FTO gene in macrophages, we demonstrated that FTO affects the stiffness-controlled macrophage inflammatory response by sustaining the negative feedback generated by SOCS1. Finally, we determined that the m 6 A reader YTHDF1 binds Socs1 mRNA and thereby maintains expression of SOCS1. Our results suggest that the FTO/Socs1/YTHDF1 regulatory axis is vital to the stiffness-controlled macrophage inflammatory response and that the deletion of FTO affects the negative feedback control exerted by SOCS1. Our findings increase understanding of the regulatory mechanisms involved in macrophage activation and the control of inflammation.

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NFAT5 Amplifies Antipathogen Responses by Enhancing Chromatin Accessibility, H3K27 Demethylation, and Transcription Factor Recruitment

Cited by 11 publications

References 64 publications

Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

Sublytic C5b-9 Induces CCL3/4 Production and Macrophage Accumulation in Thy-1N Rats via PKC-α/p65/IRF-8 Axis

NFAT5-dependent transcriptional stress control of endothelial cells prevents maladaptive remodeling of pulmonary arterioles in the hypoxic lung

N6-methyladenosine of Socs1 modulates macrophage inflammatory response in different stiffness environments

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