Integrating scRNA-seq and spatial transcriptomics to explore the implication of G6PD on immune microenvironment in lymphatic metastasis of breast cancer

Hongsen Liu ¹, Mengting Chen ², Bo Hong ³

¹Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Engineering Research Center of Oral Biomaterials and Devices of Zhejiang Province, Hangzhou, 310000, China.
²Department of Clinical Laboratory, Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Hangzhou, 310000, China.
³Department of Pathology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China.
⁴The Queen's University of Belfast Joint College, China Medical University, Shenyang, 110122, China.
⁵Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Engineering Research Center of Oral Biomaterials and Devices of Zhejiang Province, Hangzhou, 310000, China. ddmm55@zju.edu.cn.
⁶Department of Clinical Laboratory, Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Hangzhou, 310000, China. qianyun1985@zju.edu.cn.

⁰Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Engineering Research Center of Oral Biomaterials and Devices of Zhejiang Province, Hangzhou, 310000, China.

Medical Oncology +

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July 21, 2025

View abstract on PubMed

Summary

Glucose-6-phosphate dehydrogenase (G6PD) is elevated in breast cancer, promoting lymph node metastasis by fostering an immunosuppressive microenvironment. High G6PD expression correlates with poor prognosis and chemotherapy resistance.

Area Of Science

Oncology
Immunology
Molecular Biology

Background

Breast cancer metastasis, particularly to lymph nodes, is a key determinant of poor prognosis.
The mechanisms driving the immunosuppressive tumor microenvironment that facilitates lymph node metastasis are not fully understood.

Purpose Of The Study

To investigate the molecular mechanisms underlying lymph node metastasis in breast cancer.
To identify potential biomarkers for predicting breast cancer prognosis and metastasis risk.

Main Methods

Single-cell sequencing and spatial transcriptomics were employed to profile 118,127 cells from breast cancer patients and their metastatic lymph nodes.
G6PD expression levels were analyzed in tumor tissues, adjacent tissues, and metastatic lymph nodes.
Correlation analyses were performed between G6PD expression, immune cell infiltration (specifically CD4 Treg cells), clinical data, and patient prognosis.

Main Results

The pentose phosphate pathway, including Glucose-6-phosphate dehydrogenase (G6PD), was activated during breast cancer metastasis.
G6PD expression was significantly higher in breast cancer tumor tissues compared to adjacent tissues and was elevated in metastatic lymph nodes.
High G6PD expression correlated with poorer prognosis, increased CD4 Treg cell infiltration, and an immunosuppressive microenvironment in metastatic lymph nodes.
Elevated G6PD expression was also associated with lymph node metastasis, overall malignancy, and resistance to chemotherapy.

Conclusions

G6PD plays a critical role in promoting breast cancer lymph node metastasis by driving Treg cell infiltration and creating an immunosuppressive microenvironment.
G6PD serves as a potential molecular marker for predicting lymph node metastasis risk, prognosis, and chemotherapy response in breast cancer patients.

Keywords:

G6PD Pentose phosphate pathway Single-cell RNA sequencing Spatial transcriptomics