DDIT4 knockdown suppresses venous malformation progression by inhibiting NF-κB signaling as a potential therapeutic target

Yang He; Jian Lin; Yi Li; Xiaobo Cheng; Tong Wang; Wei Wang; Weixing Zeng; Yongsheng Li

doi:10.1016/j.mvr.2025.104833

DDIT4 knockdown suppresses venous malformation progression by inhibiting NF-κB signaling as a potential therapeutic target

Microvasc Res. 2025 Jun 24:104833. doi: 10.1016/j.mvr.2025.104833. Online ahead of print.

Authors

Yang He¹, Jian Lin², Yi Li³, Xiaobo Cheng⁴, Tong Wang⁵, Wei Wang⁵, Weixing Zeng⁵, Yongsheng Li⁶

Affiliations

¹ Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China; The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650031, China; Shiyan Key Laboratory of Comprehensive Prevention and Treatment of Oral Cancer, Department of Stomatology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China; Institute of Oral Diseases, School of Dentistry, Hubei University of Medicine, Shiyan, Hubei 442000, China.
² Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China; The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650031, China; Kunming Stomatological Hospital North City District Branch, 650231, China.
³ The 920th Hospital of the Joint Logistic Support Force of the Chinese People's Liberation Army, Kunming, Yunnan 650100, China.
⁴ Shiyan Key Laboratory of Comprehensive Prevention and Treatment of Oral Cancer, Department of Stomatology, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, China; Institute of Oral Diseases, School of Dentistry, Hubei University of Medicine, Shiyan, Hubei 442000, China.
⁵ Yunnan Key Laboratory of Stomatology, Kunming Medical University, Kunming, Yunnan 650500, China; The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650031, China.
⁶ Department of oral and maxillofacial surgery, Yunnan First People's Hospital, Kunming, Yunnan 650032, China; The Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan 650032, China. Electronic address: liyongshengkm@163.com.

PMID: 40571189
DOI: 10.1016/j.mvr.2025.104833

Abstract

Background: This study aims to investigate the regulatory role and underlying molecular mechanisms of DNA Damage Inducible Transcript 4 (DDIT4) in the pathogenesis of Venous Malformations (VMs), providing foundational experimental evidence for potential targeted therapies.

Methods: Bioinformatic analysis identified DDIT4 as a key differentially expressed gene in VMs, and the sgGSEA method was employed to predict its potential biological functions. Immunohistochemical staining and immunofluorescence were performed to validate the expression level of DDIT4 and its association with vascular density. A lentiviral VMs cell model was established to assess DDIT4 expression levels. The effects of DDIT4 knockdown on VMs cell function were evaluated, with mechanistic insights gained through transcriptome sequencing and Western blot analysis. Further validation was performed using 3D VMs cell models and nude mouse xenografts with DDIT4 knockdown. Additionally, exogenous functional rescue experiments were conducted by activating the NF-κB pathway with lipopolysaccharide (LPS) in DDIT4 knockdown VMs 3D cell models and nude mouse xenografts to further investigate the role of DDIT4.

Results: DDIT4 was upregulated in VMs tissues and correlated with angiogenesis. DDIT4 knockdown increased cell roundness, inhibited proliferation, migration, and NF-κB pathway activation, and blocked angiogenesis in VMs 3D models and lesion formation in nude mouse xenografts, while suppressing the NF-κB pathway in both. NF-κB pathway activation restored angiogenesis in both models.

Conclusions: DDIT4 knockdown inhibits VMs progression by suppressing the NF-κB pathway, suggesting that DDIT4 may serve as a potential therapeutic target.

Keywords: Angiogenesis; DDIT4; NF-κB pathway; Venous malformations.