NLRP12 is an innate immune checkpoint for repressing IFN signatures and attenuating lupus nephritis progression
Yen-Po Tsao, … , Chang-Youh Tsai, Szu-Ting Chen
Yen-Po Tsao, … , Chang-Youh Tsai, Szu-Ting Chen
Published February 1, 2023
Citation Information: J Clin Invest. 2023;133(3):e157272. https://doi.org/10.1172/JCI157272.
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Research Article Autoimmunity Inflammation

NLRP12 is an innate immune checkpoint for repressing IFN signatures and attenuating lupus nephritis progression

Abstract

Signaling driven by nucleic acid sensors participates in interferonopathy-mediated autoimmune diseases. NLRP12, a pyrin-containing NLR protein, is a negative regulator of innate immune activation and type I interferon (IFN-I) production. Peripheral blood mononuclear cells (PBMCs) derived from systemic lupus erythematosus (SLE) patients expressed lower levels of NLRP12, with an inverse correlation with IFNA expression and high disease activity. NLRP12 expression was transcriptionally suppressed by runt-related transcription factor 1–dependent (RUNX1-dependent) epigenetic regulation under IFN-I treatment, which enhanced a negative feedback loop between low NLRP12 expression and IFN-I production. Reduced NLRP12 protein levels in SLE monocytes was linked to spontaneous activation of innate immune signaling and hyperresponsiveness to nucleic acid stimulations. Pristane-treated Nlrp12–/– mice exhibited augmented inflammation and immune responses; and substantial lymphoid hypertrophy was characterized in NLRP12-deficient lupus-prone mice. NLRP12 deficiency mediated the increase of autoantibody production, intensive glomerular IgG deposition, monocyte recruitment, and the deterioration of kidney function. These were bound in an IFN-I signature–dependent manner in the mouse models. Collectively, we reveal a remarkable link between low NLRP12 expression and lupus progression, which suggests the impact of NLRP12 on homeostasis and immune resilience.

Authors

Yen-Po Tsao, Fang-Yu Tseng, Chih-Wei Chao, Ming-Han Chen, Yi-Chen Yeh, Babamale Olarewaju Abdulkareem, Se-Yi Chen, Wen-Ting Chuang, Pei-Ching Chang, I-Chun Chen, Pin-Hsuan Wang, Chien-Sheng Wu, Chang-Youh Tsai, Szu-Ting Chen

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Figure 2

NLRP12 promoter contains RUNX1-binding sites.

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NLRP12 promoter contains RUNX1-binding sites. (A) Sequence of human NLRP...
(A) Sequence of human NLRP12 promoter region from –751 to +222 bp. Letters in boxes denote binding sequences for RUNX1. (B) Schematic representation of NLRP12 promoter luciferase reporter constructs. For promoter analysis, an 830 bp length of NLRP12 promoter was cloned into pGL4-vector to drive luciferase reporter expression (NLRP12-Luc#1). Deletion constructs of NLRP12 promoter cloned into pGL4 vector are shown. Vertical lines are denoted as the RUNX1-binding motif on the NLRP12 promoter. (C) HEK293T cells were transfected with NLRP12-Luc#1 to NLRP12-Luc#4 plasmid and the internal control plasmid. Relative luciferase activity (rel. luc act.) was determined at 24 hours after transfection. (D) HEK293T cells transfected with NLRP12-Luc#1 and empty vector (EV) (pCDNA3) or RUNX-encoding plasmid (pCDNA3/DDK-RUNX1; 30, 100, 300, 500 ng/sample) and cell lysates were subjected to measurement of luciferase activity at 24 hours. (E and F) Human HT1080 and HEK293T cells were transfected with NLRP12-Luc#1 to NLRP12-Luc#4 for 6 hours, followed by IFN-α2 or VSV stimulation. Luciferase assays were performed at 24 hours. (G) Knockout of RUNX1 in THP-1 cells by CRISPR Cas9/sgRNA. (H) THP-1 cells with scrambled sgRNA or sgRNA targeting RUNX1 were treated with IFN-α2 or infected with VSV for 8 hours. NLRP12 expression was measured. (C–F) One-way ANOVA test (multiple samples to a control); (H) 2-tailed Student’s t test. Data are represented as mean ± SEM (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001.