NLRP12 downregulates the Wnt/β-catenin pathway via interaction with STK38 to suppress colorectal cancer
Shahanshah Khan, … , Thirumala-Devi Kanneganti, Hasan Zaki
Shahanshah Khan, … , Thirumala-Devi Kanneganti, Hasan Zaki
Published August 15, 2023
Citation Information: J Clin Invest. 2023;133(19):e166295. https://doi.org/10.1172/JCI166295.
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Research Article Gastroenterology

NLRP12 downregulates the Wnt/β-catenin pathway via interaction with STK38 to suppress colorectal cancer

Abstract

Colorectal cancer (CRC) at advanced stages is rarely curable, underscoring the importance of exploring the mechanism of CRC progression and invasion. NOD-like receptor family member NLRP12 was shown to suppress colorectal tumorigenesis, but the precise mechanism was unknown. Here, we demonstrate that invasive adenocarcinoma development in Nlrp12-deficient mice is associated with elevated expression of genes involved in proliferation, matrix degradation, and epithelial-mesenchymal transition. Signaling pathway analysis revealed higher activation of the Wnt/β-catenin pathway, but not NF-κB and MAPK pathways, in the Nlrp12-deficient tumors. Using Nlrp12–conditional knockout mice, we revealed that NLRP12 downregulates β-catenin activation in intestinal epithelial cells, thereby suppressing colorectal tumorigenesis. Consistent with this, Nlrp12-deficient intestinal organoids and CRC cells showed increased proliferation, accompanied by higher activation of β-catenin in vitro. With proteomic studies, we identified STK38 as an interacting partner of NLRP12 involved in the inhibition of phosphorylation of GSK3β, leading to the degradation of β-catenin. Consistently, the expression of NLRP12 was significantly reduced, while p-GSK3β and β-catenin were upregulated in mouse and human colorectal tumor tissues. In summary, NLRP12 is a potent negative regulator of the Wnt/β-catenin pathway, and the NLRP12/STK38/GSK3β signaling axis could be a promising therapeutic target for CRC.

Authors

Shahanshah Khan, Youn-Tae Kwak, Lan Peng, Shuiqing Hu, Brandi L. Cantarel, Cheryl M. Lewis, Yunpeng Gao, Ram S. Mani, Thirumala-Devi Kanneganti, Hasan Zaki

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Figure 3

Increased tumorigenesis of Nlrp12–/– mice is associated with higher β-catenin activation in the tumor.

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Increased tumorigenesis of Nlrp12–/– mice is associated with higher β-ca...
WT and Nlrp12–/– mice were separately housed (SH) (n = 6/group) or cohoused (CH) (n = 7/group). Colorectal tumorigenesis was induced by AOM plus DSS treatment. Mice were sacrificed 12 weeks after AOM injection. (A–D) Tumors were analyzed for the activation of indicated signaling pathways by Western blotting. Band intensities of p-P65, p-IκBa, and p-ERK were measured densitometrically. Data represent mean ± SEM. (E–H) The expression of β-catenin, cMyc, and cyclin D1 in tumors was analyzed by Western blotting. Band intensities of β-catenin, cMyc, and cyclin D1 were measured densitometrically. Data represent mean ± SEM. (I and J) Colon sections were stained for Ki67 and positive cells (red) were counted under 20× objective (n = 3 mice/group). Scale bars: 200 μm. Data represent mean ± SEM. (K) Tumor-bearing colon sections were immunostained for β-catenin (red). The nuclei were stained with DAPI (blue). Scale bars: 200 μm. All experiments were repeated at least 2 times, and data from a representative experiment are presented. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by unpaired, 2-tailed Student’s t test.