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Integrated Single-Cell and Bulk RNA Sequencing Identifies SERPING1 as a Biomarker of Immune Infiltration and

Yuhang Shang¹, Runze Guo², Jiangwei Liu¹

¹Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, 150081 Harbin, Heilongjiang, China.

Frontiers in Bioscience (Landmark Edition)

|January 29, 2026

Abstract

Background:

Triple-negative breast cancer (TNBC) is an aggressive malignancy that lacks effective treatment. Immune infiltration plays an important role in anti-tumor responses. Serpin family G1 (SERPING1), a biomarker associated with immune infiltration, has been implicated in multiple cancers, but its role in TNBC remains unclear.

Methods:

RNA sequencing and clinical data for TNBC were obtained from the Gene Expression Omnibus, the Cancer Genome Atlas, and the Molecular Taxonomy of Breast Cancer International Consortium databases. First, the expression, prognostic value, and biological functions of SERPING1 were analyzed. Then, the tumor microenvironment (TME) was comprehensively characterized, and the relationship between SERPING1 expression and immunotherapy response was assessed. Immunohistochemical staining was performed to confirm SERPING1 expression and the abundance of CD4+ T cells and CD8+ T cells in clinical specimens. Finally, single-cell analysis was conducted to investigate the role of SERPING1 in immune cell activation.

Results:

SERPING1 was downregulated in TNBC and was an independent predictor of survival. Functionally, SERPING1 activated the immune response in TNBC patients. Mechanistically, elevated SERPING1 levels lead to increased immune cell infiltration, particularly of CD4+ and CD8+ T cells, in the TME. Moreover, SERPING1 was primarily localized in cancer-associated fibroblasts (CAFs), with SERPING1+ apCAFs exhibiting increased communications with anti-tumor immune cells at the single-cell level.

Conclusions:

SERPING1 contributes to enhanced immune cell infiltration, desirable immunotherapy response, and improved prognosis. It thus can be utilized as a promising biomarker for immune infiltration and prognosis. These findings provide novel insights into TME-related immune regulation and may inform strategies to enhance immunotherapy efficacy in TNBC.

Keywords:

biomarker immunomodulation immunotherapy prognosis triple negative breast neoplasms tumor microenvironment

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