Radiochemotherapy (RCT) is a powerful treatment for cervical cancer, which affects not only malignant cells but also the immune and stromal compartments of the tumor. Understanding the remodeling of the local ecosystem induced by RCT would provide valuable insights into improving treatment strategies for cervical cancer. In this study, we applied single-cell RNA-sequencing to paired pre- and post-RCT tumor biopsies from patients with cervical cancer and adjacent normal cervical tissues. We found that the residual population of epithelial cells post-RCT showed upregulated expression of MHC class II genes. Moreover, RCT led to the accumulation of monocytic myeloid-derived suppressor cells with increased pro-inflammatory features and CD16+ NK cells with a higher cytotoxic gene expression signature. However, subclusters of T cells showed no significant increase in the expression of cytotoxic features post-RCT. These results reveal the complex responses of the tumor ecosystem to RCT, providing evidence of activation of innate immunity and MHC-II upregulation in cervical cancer.

The intratumor heterogeneity of human papillomavirus (HPV)‐related cervical cancer remains poorly defined. We performed single‐cell RNA sequencing on 18 046 individual cells derived from two HPV‐related cervical adenosquamous carcinoma samples to analyze the transcriptional heterogeneity of both epithelial and immune constituents, identifying seven epithelial (Epi1‐7) and 11 immune subclusters. Based on expression of known cervical cancer markers, Epi1‐2 primarily displayed features of adenocarcinoma, whereas Epi3‐6 were instead characterized by features of squamous carcinoma. Our analyses also revealed that hypoxia and Kirsten rat sarcoma viral oncogene signaling were highly represented within Epi1; metabolic pathways mediating glycolysis and oxidative phosphorylation were enriched in Epi2‐4; while Epi5 was enriched in p53 pathway components and features of epithelial–mesenchymal transition. Moreover, CD8+FGFBP2+ T cells and FGFBP2+ natural killer cells were found to display high levels of cytotoxic effectors (GZMA, GZMB, GNLY, and PRF1) and low levels of inhibitory markers (PDCD1, TIGIT, and CTLA4), such that tumor infiltration by these populations was positively associated with survival in a cohort of n = 165 patients with HPV‐related cervical cancer from The Cancer Genome Atlas database (p = 0.017 and 0.014, respectively). These results shed new light on the intratumor heterogeneity of HPV‐related cervical adenosquamous carcinoma, which will help to refine diagnostic and treatment approaches.

Dear Editor,Increasing numbers of studies have revealed the immunomodulating effects of radiotherapy when combined with immune checkpoint inhibitors, as radioimmunotherapy has proven to be a promising treatment [1]. Radioimmunotherapy has shown significantly improved tumor responses than radiotherapy or immunotherapy alone in various malignant tumors [2][3][4]. It has also been applied to cervical cancer in multiple ongoing clinical trials (NCT03612791 [5] and NCT02635360 [6]). However, tumor recurrence and metastasis are often unavoidable. As such, investigations into radioimmunotherapy-induced tumor ecosystem evolution are essential for guiding improvements in treatment strategies that achieve better long-term disease control. To date, several studies have investigated radiochemotherapy-induced tumor ecosystem evolution using bulk RNA-sequencing and immune staining [7,8]. However, these findings were limited owing to the cellular heterogeneity in cancers. Single-cell RNA-sequencing (scRNA-seq) enables the characterization of cell compositions and transcriptomic states in the tumor at single-cell resolution.To investigate radioimmunotherapy-induced intratumoral changes, we performed scRNA-seq on a pair of cervical squamous cell carcinoma (CESC) samples before and during radioimmunotherapy. The study protocols are found in the Supplementary Materials. A total of 17,769 cells were obtained, with an average gene number of 1,875 after quality control. Using uniform manifold approximation and projection (UMAP) analysis, we identified nine main cell types: T cells (CD3D + , CD3E + ), plasma cells (IGHG1 + , IGHG3 + ), macrophages (CD14 + , CD68 + ), monocytic myeloid-derived suppressor cells (M-MDSCs