NEDD4L mediates intestinal epithelial cell ferroptosis to restrict inflammatory bowel diseases and colorectal tumorigenesis
Jingjing Liang, … , Xiaojian Wang, Wenlong Lin
Jingjing Liang, … , Xiaojian Wang, Wenlong Lin
Published December 17, 2024
Citation Information: J Clin Invest. 2025;135(3):e173994. https://doi.org/10.1172/JCI173994.
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Research Article Cell biology Inflammation

NEDD4L mediates intestinal epithelial cell ferroptosis to restrict inflammatory bowel diseases and colorectal tumorigenesis

Abstract

Various factors play key roles in maintaining intestine homeostasis. Disruption of the balance may lead to inflammatory bowel diseases and even colorectal cancer (CRC). Loss or gain of function of many key proteins can result in dysregulated intestinal homeostasis. Our research demonstrated that neural precursor cells expressed developmentally downregulated 4–like protein (NEDD4L, or NEDD4-2), a type of HECT family E3 ubiquitin ligase, played an important role in maintaining intestinal homeostasis. NEDD4L expression was significantly inhibited in intestinal epithelial cells (IECs) of patients with Crohn’s disease, ulcerative colitis, and CRC. Global KO of NEDD4L or its deficiency in IECs exacerbated colitis induced by dextran sulfate sodium (DSS) and 2,4,6-trinitrobenzene sulfonic acid (TNBS) and CRC induced by azoxymethane and DSS. Mechanistically, NEDD4L deficiency in IECs inhibited expression of the key ferroptosis regulator glutathione peroxidase 4 (GPX4) by reducing the protein expression of solute carrier family 3 member 2 (SLC3A2) without affecting its gene expression, ultimately promoting DSS-induced IEC ferroptosis. Importantly, ferroptosis inhibitors reduced the susceptibility of NEDD4L-deficient mice to colitis and colitis-associated CRC. Thus, NEDD4L is an important regulator in IEC ferroptosis, maintaining intestinal homeostasis, making it a potential clinical target for diagnosing and treating IBDs.

Authors

Jingjing Liang, Ning Wang, Yihan Yao, Yingmei Wang, Xiang An, Haofei Wang, Huan Liu, Yu Jiang, Hui Li, Xiaoqing Cheng, Jiaqi Xu, Xiaojing Liang, Jun Lou, Zengfeng Xin, Ting Zhang, Xiaojian Wang, Wenlong Lin

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Figure 6

SLC3A2 negatively regulates ferroptosis.

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SLC3A2 negatively regulates ferroptosis. (A–C) The multiple cell lines, ...
(A–C) The multiple cell lines, including HCT116 cells (A), SW480 cells (B), and RKO cells (C), were transfected with siRNA targeted to SLC3A2 (siSLC3A2) or negative control (siNC). The cells were treated with 2% DSS for the indicated times and then subjected to CCK8 assay. (D–F) The multiple cell lines were treated as in A–C with or without Fer-1 (2 μM) treatment. The cells were then subjected to flow cytometry analysis of BODIPY C11 staining to measure lipid peroxidation production. (G–I) The multiple cell lines were treated as in A–C for the indicated times and then subjected to immunoblot analysis of the indicated proteins. (J–M) FLAG-tagged SLC3A2 or FLAG-tagged null control plasmids were overexpressed in HCT116 cells. The cells were treated with 2% DSS or indicated inducers for the stated times, and then subjected to CCK8 assay (J), MDA assay (K), flow cytometry analysis of BODIPY C11 staining (L), and immunoblot analysis of the indicated proteins (M). (N–P) HCT116 cells were transfected with oligonucleotides specific for siRNA negative control (siNC) or NEDD4L (siNEDD4L), and then FLAG-tagged SLC3A2 or FLAG-tagged null control plasmid was overexpressed in the cells. The cells were treated with 2% DSS for the indicated times and then subjected to CCK8 assay (N) and lipid peroxidation (O). (P) Immunoblot analysis of the indicated proteins. Data represent mean ± SEM from at least 2 independent experiments. Each dot represents an independent sample. *P < 0.05; **P < 0.01; ***P < 0.001. Statistical analysis was performed using a 2-tailed Student’s t test in A–F, J–L, N, and O.