Single-nucleus RNA Sequencing and Spatial Transcriptomics Reveal the Immunological Microenvironment of Cervical Squamous Cell Carcinoma

Abstract

To understand the etiological, structural, and immunological characteristics of cervical squamous cell carcinoma (CSCC), we conducted single nucleus RNA sequencing (snRNA-seq) and spatial transcriptomics (ST) experiments for cervical samples from 20 individuals. When exploring the possible factors shaping the intra-individual immune heterogeneity in CSCC, we identified a cluster of cancer-associated fibroblasts (CAFs) enriched around some tumors, which highly expressed ACTA2, POSTN, ITGB4, and FAP. Results showed that the CAFs might support the growth and metastasis of tumors by inhibiting lymphocyte infiltration and remodeling the tumor extracellular matrix. Moreover, high CAF signals predicted poorer clinical outcomes for CSCC patients. Our data also revealed the infection profiles of HPV in tumors, the critical factors involved in the progression of cervical cancerous lesions, and the association between tumor metabolism and immune response intensity. Collectively, our findings may improve the prognostic and therapeutic methods for CSCC.

Improved Protocol for Single-Nucleus RNA-sequencing of Frozen Human Bladder Tumor Biopsies

“…A third study on human cervical squamous cell carcinoma also identified a diverse population of immune cells 17 . Although they identify immune cells, these studies might underestimate the true fraction of immune cells present in their natural conditions after all.…”

“…However, in another study, Gouin et al performed snRNA-seq on fresh frozen bladder tumor samples and identified a fraction of immune cells (5%) comprising T-cells, dendritic cells, macrophages, and B-cells defined by classic immune marker genes 4 . A third study on human cervical squamous cell carcinoma also performing snRNA-seq identified a diverse population of immune cells as well 18 . Although they identify immune cells, these studies might underestimate the true fraction of immune cells present in their natural conditions after all.…”

Re‐exploring tissue regeneration by novel spatial transcriptomics technologies

“…Ou et al. found unique immune cell types enriched in tumour areas with different metabolic activity 9 . By analyzing neighboring cell composition from spatial transcriptomics data, Cong et al.…”

“…8 Ou et al found unique immune cell types enriched in tumour areas with different metabolic activity. 9 By analyzing neighboring cell composition from spatial transcriptomics data, Cong et al identified a specific macrophage subgroup for SARS-CoV-2 clearance and inflammation resolution 10 (Figure 1).…”

Editorial: Harnessing tumor microenvironment for gynecologic cancer therapy

“…Consequently, the TME has the potential to switch the immune system from an anti-tumor mode to a pro-tumor state, depending on its composition ( Figure 1 ). Encouragingly, therapeutic approaches targeting TME components, including myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), and regulatory T cells (Tregs), and have demonstrated promising anti-tumor activity in both preclinical and clinical studies ( 12 – 18 ). Therefore, exploring the predictive and therapeutic values of the TME holds significant promise for advancing gynecologic cancer treatment.…”