Helicobacter pylori SlyD stabilizes TPT1 via hnRNPK and enhances OCT1-mediated CDX2 transcriptional activation to drive gastric intestinal metaplasia

Shuwen Zheng; Yingying Wang; Chuxuan Ni; Rui Guo; Xunan Qiu; Jijun Chen; Lu Wang; Xiaohu Sun; Moye Chen; Yunen Liu; Yuan Yuan; Yuehua Gong

doi:10.1186/s12916-025-03911-8

Helicobacter pylori SlyD stabilizes TPT1 via hnRNPK and enhances OCT1-mediated CDX2 transcriptional activation to drive gastric intestinal metaplasia

BMC Med. 2025 Feb 6;23(1):71. doi: 10.1186/s12916-025-03911-8.

Authors

Shuwen Zheng^{1

2

3}, Yingying Wang^{1

2

3}, Chuxuan Ni^{1

2

3}, Rui Guo^{1

2

3}, Xunan Qiu^{1

2

3}, Jijun Chen^{1

2

3}, Lu Wang^{1

2

3}, Xiaohu Sun^{1

2

3}, Moye Chen^{1

2

3}, Yunen Liu^{1

2

3}, Yuan Yuan^{4

5

6}, Yuehua Gong^{7

8

9}

Affiliations

¹ Tumor Etiology and Screening Department of Cancer Institute and General Surgery, the First Hospital of China Medical University, Shenyang, 110001, China.
² Key Laboratory of Cancer Etiology and Prevention in Liaoning Education Department, the First Hospital of China Medical University, Shenyang, 110001, China.
³ Key Laboratory of GI Cancer Etiology and Prevention in Liaoning Province, the First Hospital of China Medical University, Shenyang, 110001, China.
⁴ Tumor Etiology and Screening Department of Cancer Institute and General Surgery, the First Hospital of China Medical University, Shenyang, 110001, China. yuanyuan@cmu.edu.cn.
⁵ Key Laboratory of Cancer Etiology and Prevention in Liaoning Education Department, the First Hospital of China Medical University, Shenyang, 110001, China. yuanyuan@cmu.edu.cn.
⁶ Key Laboratory of GI Cancer Etiology and Prevention in Liaoning Province, the First Hospital of China Medical University, Shenyang, 110001, China. yuanyuan@cmu.edu.cn.
⁷ Tumor Etiology and Screening Department of Cancer Institute and General Surgery, the First Hospital of China Medical University, Shenyang, 110001, China. yhgong@cmu.edu.cn.
⁸ Key Laboratory of Cancer Etiology and Prevention in Liaoning Education Department, the First Hospital of China Medical University, Shenyang, 110001, China. yhgong@cmu.edu.cn.
⁹ Key Laboratory of GI Cancer Etiology and Prevention in Liaoning Province, the First Hospital of China Medical University, Shenyang, 110001, China. yhgong@cmu.edu.cn.

Abstract

Background: Gastric intestinal metaplasia (GIM) represents an important precancerous lesion in intestinal-type gastric cancer, triggered by persistent Helicobacter pylori (H. pylori) infection. In a previous study, we unveiled SlyD as a novel virulence factor of H. pylori, establishing its role in GIM induction through TPT1. However, the underlying mechanism remains undetermined.

Methods: Gastric epithelial cells were stimulated with H. pylori 26695, a SlyD inactivated mutant (ΔSlyD), and purified HpSlyD protein, respectively. Real-time qPCR and western blot were subsequently used to assess the expression levels of hnRNPK, TPT1, OCT1, and GIM markers. RNA sequencing was employed to identify differentially expressed genes associated with H. pylori SlyD infection. Protein stability was evaluated using cycloheximide. Molecular interactions were investigated through co-immunoprecipitation, chromatin immunoprecipitation, and dual-luciferase reporter assays. Additionally, molecular docking was utilized to predict TPT1 inhibitors. Immunohistochemistry staining was conducted to validate hnRNPK, TPT1, OCT1, and CDX2 expression in gastric tissue samples from both human and Mongolian gerbils.

Results: H. pylori SlyD upregulates TPT1 and induces the expression of GIM markers through hnRNPK. The interaction between hnRNPK and TPT1 enhances TPT1 protein stability, with H. pylori SlyD intensifying this association. TPT1 promotes the expression of GIM markers mediated via OCT1, which binds to CDX2 promoter region, thereby modulating its transcriptional activity. Dihydroartemisinin has the potential to target TPT1, inhibiting the H. pylori SlyD-induced expression of GIM markers.

Conclusions: In vitro and in vivo experiments verified that H. pylori SlyD enhances TPT1 stability through hnRNPK, leading to OCT1-mediated transcriptional activation of CDX2 and the initiation of the GIM process. Our study offers novel perspectives on the pathogenesis of H. pylori-related gastric precancerous conditions.

Keywords: H. pylori; CDX2; Dihydroartemisinin; Gastric intestinal metaplasia; Gastric precancerous disease; SlyD.

MeSH terms

Animals
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
CDX2 Transcription Factor* / genetics
CDX2 Transcription Factor* / metabolism
Gastric Mucosa / metabolism
Gastric Mucosa / microbiology
Gastric Mucosa / pathology
Gerbillinae
Helicobacter Infections / genetics
Helicobacter Infections / metabolism
Helicobacter Infections / microbiology
Helicobacter Infections / pathology
Helicobacter pylori*
Humans
Male
Metaplasia* / genetics
Metaplasia* / metabolism
Stomach Neoplasms / genetics
Stomach Neoplasms / metabolism
Stomach Neoplasms / microbiology
Stomach Neoplasms / pathology
Transcriptional Activation
Tumor Protein, Translationally-Controlled 1*

Substances

CDX2 Transcription Factor
Tumor Protein, Translationally-Controlled 1
Bacterial Proteins
CDX2 protein, human
TPT1 protein, human

Abstract

MeSH terms

Substances

Grants and funding