NOX4 Suppresses Ferroptosis Through Regulation of the Pentose Phosphate Pathway in Colorectal Cancer

Abstract

Objective

Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOXs) are known as major sources of reactive oxygen species (ROS), yet their role in regulating cellular antioxidative metabolism and ferroptosis is unclear. This study assessed the expression and clinical relevance of NOXs across pan-cancer and investigated the role of NOX4 in colorectal cancer progression

Methods

We analyzed transcriptomic and survival data from The Cancer Genome Atlas (TCGA) for NOXs across 22 types of solid tumors. A CRISPR library targeting NOXs was developed for potential therapeutic target screening in colorectal cancer cells (CRCs). Techniques such as CRISPR-knockout cell lines, 1,2-13C-glucose tracing, PI staining, BrdU assays, and coimmunoprecipitation were employed to elucidate the function of NOX4 in CRCs.

Results

NOX4 emerged as a key therapeutic target for colorectal cancer from TCGA data. CRISPR screening highlighted its essential role in CRC survival, with functional experiments confirming that NOX4 upregulation promotes cell survival and proliferation. The interaction of NOX4 with glucose-6-phosphate dehydrogenase (G6PD) was found to enhance the pentose phosphate pathway (PPP), facilitating ROS clearance and protecting CRCs against ferroptosis.

Conclusions

This study identified NOX4 as a novel ferroptosis suppressor and a therapeutic target for the treatment of colorectal cancer. The findings suggest that a coupling between NADPH oxidase enzyme NOX4 and the PPP regulates ferroptosis and reveal an accompanying metabolic vulnerability for therapeutic targeting in colorectal cancer.

Keywords

Ferroptosis / Pentose phosphate pathway / Oxidative stress / Colorectal cancer

© The Author(s), under exclusive licence to Huazhong University of Science and Technology 2025

Supplementary Information The online version contains supplementary material available at https:// doi. org/ 10. 1007/ s11596-025-00013-7.

Acknowledgements The authors acknowledge all those who contributed to The Cancer Genome Atlas Program.

Author Contributions Jing Zhu: Conceptualization; Resources; Formal analysis; Supervision; Funding acquisition; Visualization; Project administration. Writing—original draft. Chao Jiang: Data curation; Formal analysis; Investigation; Methodology. Fan Wang: Formal analysis; Data curation; Investigation; Methodology. Ming-Yue Tao: Data curation; Investigation; Methodology. Hai-xiao Wang: Data curation; Investigation; Methodology. Yuan Sun: Data curation; Investigation; Methodology. Hong-xia Hui: Resources; Formal analysis; Visualization, Investigation; Visualization; Project administration; Writing— review & editing.

Funding This work was supported by the National Natural Science Foundation of China (No. 81802917 and No. 82102969) and the Huai’an Natural Science Research Program (No. HAB202101).

Availability of Data and Materials The 22 cancer-type data were derived from the TCGA Research Network: http://cancergenome. nih.gov/. Analysis data of gene expression alterations of NOX family enzymes in 22ccancer types can be found in the supplemental materials. The other datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Declarations

Conflict of interest The authors declare that they have not competing interests.

Ethics Approval and Consent to Participate Not applicable.

Consent for publication Not applicable.

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