HOXD10 attenuates renal fibrosis by inhibiting NOX4-induced ferroptosis

• Published in: Cell death & disease 2024-06, Vol.15 (6), p.398-14
• Main Authors: Li, Xin, Ma, Tian-Kui, Wang, Pu, Shi, Hang, Hai, Sang, Qin, Yu, Zou, Yun, Zhu, Wan-Ting, Li, Hui-Min, Li, Yan-Nong, Yin, Li, Xu, Yan-Yan, Yang, Qi, Zhang, Shuang, Ding, Hong
• Format: Article
• Language: English
• Subjects: 13/1; 13/2; 42/35; 42/41; 45/41; 631/80/82; 64/60; 692/699/1585/104; 96/44; Animal models; Animals; Antibodies; Biochemistry; Biomedical and Life Sciences; Bisulfite; Cell Biology; Cell Culture; Cell Line; Chromatin; Disease Models, Animal; Epithelial cells; Ferroptosis; Ferroptosis - genetics; Fibrosis; Gene expression; Glutathione; Homeobox; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humans; Immunology; Immunoprecipitation; Kidney - metabolism; Kidney - pathology; Kidney diseases; Life Sciences; Male; Malondialdehyde; Medical innovations; Mice; Mice, Inbred C57BL; NADPH Oxidase 4 - genetics; NADPH Oxidase 4 - metabolism; NOX4 protein; Oxidative Stress; Plasmids; Reactive oxygen species; Reactive Oxygen Species - metabolism; Renal function; Renal Insufficiency, Chronic - genetics; Renal Insufficiency, Chronic - metabolism; Renal Insufficiency, Chronic - pathology; Superoxide dismutase; Transcription activation; Transcription Factors - genetics; Transcription Factors - metabolism; Transforming Growth Factor beta1 - metabolism; Transforming growth factor-b1
• ISSN: 2041-4889; 2041-4889
• DOI: 10.1038/s41419-024-06780-w

In chronic kidney disease (CKD), renal fibrosis is an unavoidable result of various manifestations. However, its pathogenesis is not yet fully understood. Here, we revealed the novel role of Homeobox D10 (HOXD10) in CKD-related fibrosis. HOXD10 expression was downregulated in CKD-related in vitro and in vivo fibrosis models. UUO model mice were administered adeno-associated virus (AAV) containing HOXD10, and HOXD10 overexpression plasmids were introduced into human proximal tubular epithelial cells induced by TGF-β1. The levels of iron, reactive oxygen species (ROS), lipid ROS, the oxidized glutathione/total glutathione (GSSG/GSH) ratio, malonaldehyde (MDA), and superoxide dismutase (SOD) were determined using respective assay kits. Treatment with AAV–HOXD10 significantly attenuated fibrosis and renal dysfunction in UUO model mice by inhibiting NOX4 transcription, ferroptosis pathway activation, and oxidative stress. High levels of NOX4 transcription, ferroptosis pathway activation and profibrotic gene expression induced by TGF-β1/erastin (a ferroptosis agonist) were abrogated by HOXD10 overexpression in HK-2 cells. Moreover, bisulfite sequencing PCR result determined that HOXD10 showed a hypermethylated level in TGF-β1-treated HK-2 cells. The binding of HOXD10 to the NOX4 promoter was confirmed by chromatin immunoprecipitation (ChIP) analysis and dual-luciferase reporter assays. Targeting HOXD10 may represent an innovative therapeutic strategy for fibrosis treatment in CKD.