Dissecting the Single-Cell Transcriptome Network of Immune Environment Underlying Cervical Premalignant Lesion, Cervical Cancer and Metastatic Lymph Nodes

Abstract

Cervical cancer (CC) is one of the most common malignancy in women worldwide. It is characterized by a natural continuous phenomenon, that is, it is in the initial stage of HPV infection, progresses to intraepithelial neoplasia, and then develops into invasion and metastasis. Determining the complexity of tumor microenvironment (TME) can deepen our understanding of lesion progression and provide novel therapeutic strategies for CC. We performed the single-cell RNA sequencing on the normal cervix, intraepithelial neoplasia, primary tumor and metastatic lymph node tissues to describe the composition, lineage, and functional status of immune cells and mesenchymal cells at different stages of CC progression. A total of 59913 single cells were obtained and divided into 9 cellular clusters, including immune cells (T/NK cells, macrophages, B cells, plasma cells, mast cells and neutrophils) and mesenchymal cells (endothelial cells, smooth muscle cells and fibroblasts). Our results showed that there were distinct cell subpopulations in different stages of CC. High-stage intraepithelial neoplasia (HSIL) tissue exhibited a low, recently activated TME, and it was characterized by high infiltration of tissue-resident CD8 T cell, effector NK cells, Treg, DC1, pDC, and M1-like macrophages. Tumor tissue displayed high enrichment of exhausted CD8 T cells, resident NK cells and M2-like macrophages, suggesting immunosuppressive TME. Metastatic lymph node consisted of naive T cell, central memory T cell, circling NK cells, cytotoxic CD8+ T cells and effector memory CD8 T cells, suggesting an early activated phase of immune response. This study is the first to delineate the transcriptome profile of immune cells during CC progression using single-cell RNA sequencing. Our results indicated that HSIL exhibited a low, recently activated TME, tumor displayed immunosuppressive statue, and metastatic lymph node showed early activated phase of immune response. Our study enhanced the understanding of dynamic change of TME during CC progression and has implications for the development of novel treatments to inhibit the initiation and progression of CC.

Integrated single-cell transcriptome analysis of the tumor ecosystems underlying cervical cancer metastasis

“…Recently, an increasing number of scRNA-seq studies have indicated that macrophages cannot be simply divided into M1 and M2 macrophages (42,43). Our previous study reported that macrophages from a tumor or metastatic LNs can be defined as C1QA+ macrophages, which had a dominant M2-like phenotype and immunosuppressive function (12). However, we cannot distinguish their differences in primary tumor and metastatic LNs.…”

“…Meanwhile, we also elucidated the characteristics of CC TME, from precancerous lesions to invasive tumors, and then to metastatic lymph nodes. However, the mechanism of malignant cell metastasis is still lacking (11,12). Previous scRNA-seq studies have reported that metastatic cancer cells highly express genes related to epithelialmesenchymal transition (EMT), oxidative stress, the proteasome, and cancer stem cell biomarkers (13)(14)(15).…”

Reduced MHC Class I and II Expression in HPV−Negative vs. HPV−Positive Cervical Cancers

“…CD68 is upregulated in a proinflammatory environment, particularly by IFNγ which was seen to be expressed at higher levels in HPV+ CC ( Figure 3 ). Furthermore, previous studies using single-cell RNA-seq show that macrophage infiltration into CC tumors is relatively low compared to other cell types, such as T and NK cells [ 89 ]. These findings indicate that while both macrophages and epithelial cells may be contributing to MHC class II expression in the CC TME, the bulk of their expression is provided by the far more prevalent epithelial-derived tumor cells, as reported by IHC studies.…”

Development and validation of an autophagy-related long non-coding RNA prognostic signature for cervical squamous cell carcinoma and endocervical adenocarcinoma

“…In another aspect, rapid tumor growth and lymph node metastasis are closely related to the reversion of the CD4 + /CD8 + ratio and the reduced proportion of tumor-infiltrating CD4 + T cells in patients with cervical cancer ( 34 ). It has been shown that there were low levels of CD8 + Tem in cancerous tissue but high levels in lymph nodes and blood ( 35 ). Moreover, Tcm retains migration properties of naive T cells and has the ability to directly infiltrate the non-lymphoid tissues ( 36 ).…”