Profiling of immune cell subsets and functional characteristics of cervical cancer based on single cell RNA sequencing

A comprehensive characterization of immune cells within the tumor microenvironment (TME) of cervical squamous cell carcinoma (CSCC) is essential to advance understanding of tumor biology and guide immunotherapy development. Using single-cell RNA sequencing, we analyzed 14,441 immune cells isolated from tumor tissues and paratumor tissues of three CSCC patients. This analysis identified nine major immune cell subsets, including CD8+ T cells, regulatory T cells (Tregs), natural killer/T cells (NK/T cells), B cells, plasmacytoid dendritic cells (pDCs), and macrophages. Notably, elevated CCR7 expression in exhausted CD8+ T cells was associated with improved prognosis, suggesting it as a preliminary candidate for an immunoregulatory target that warrants further investigation. Additionally, CCL5 levels were elevated in Tregs, indicating a possible involvement in their recruitment to the tumor site that requires further validation. By integrating data on tumor suppressors, oncogenic factors, cytokines, and chemokines, we performed differential gene expression analyses across immune populations, identifying candidate genes for further mechanistic investigation. This systematic comparison of the TME between tumor and paratumor tissues reveals dynamic changes in the immune landscape, providing insights and suggesting potential targets for further study to enhance understanding and treatment of CSCC. Overall design: Fresh squamous cervical cancer specimens were collected from three patients undergoing primary surgery at the Department of Obstetrics and Gynecology, First Hospital of Jilin University, between June 2019 and August 2020. None received preoperative treatment.Samples of cervical squamous cell carcinoma (CSCC), both tumor and paratumor, were procured from the tumor mass and regions located 1 cm away from the tumor boundary, respectively. The tissue samples were washed with PBS (CORNING) before being stored in MACS Tissue Storage Solution (Miltenyi Biotec). Subsequently, they were processed into single-cell suspensions.