# Single-Cell Transcriptomic Landscape of Cervical Cancer Cell Lines Before and After Chemoradiotherapy

## TL;DR

This study uses single-cell analysis to uncover how cervical cancer cells become resistant to chemoradiotherapy, revealing complex changes in gene activity and cell types that could guide new treatments.

## Contribution

The study introduces a unique isogenic cell line model to reveal multilevel adaptations in cervical cancer chemoradioresistance.

## Key findings

- Resistance involves transcriptional reprogramming toward embryonic stemness (HOX, POU5F1) and shifts in collagen expression.
- Resistant cells show a pro-tumorigenic senescent-like population and progenitor-like reservoirs linked to Wnt, TGFβ, and BMP pathways.
- Metabolic vulnerabilities in senescent cells and specific ligands (EREG, SEMA3C) are identified as potential therapeutic targets.

## Abstract

What are the main findings?
We established and characterized a unique pair of isogenic cervical cancer cell lines (AdMer35/AdMer43) from a patient with squamous cell carcinoma of the cervix before and after chemoradiotherapy, revealing that resistance is not caused by a single factor but by complex reorganization of cancer cells.Single-cell analysis identified key resistance hallmarks: transcriptional reprogramming towards embryonic stemness (HOX, POU5F1), a shift from fibrillar to non-fibrillar collagen expression, and activation of interferon/inflammatory pathways.

We established and characterized a unique pair of isogenic cervical cancer cell lines (AdMer35/AdMer43) from a patient with squamous cell carcinoma of the cervix before and after chemoradiotherapy, revealing that resistance is not caused by a single factor but by complex reorganization of cancer cells.

Single-cell analysis identified key resistance hallmarks: transcriptional reprogramming towards embryonic stemness (HOX, POU5F1), a shift from fibrillar to non-fibrillar collagen expression, and activation of interferon/inflammatory pathways.

What are the implications of the main findings?
The study maps a remodeled tumor ecosystem with a metabolically reprogrammed, pro-tumorigenic senescent-like cell population and a progenitor-like cell reservoir, identifying them as critical therapeutic targets for eliminating resistant cell pools.The identified key pathways, Wnt, TGFβ, BMP, and the specific ligands EREG and SEMA3C, along with the metabolic vulnerabilities of the senescent compartment, provide a robust framework for future therapeutic exploration.

The study maps a remodeled tumor ecosystem with a metabolically reprogrammed, pro-tumorigenic senescent-like cell population and a progenitor-like cell reservoir, identifying them as critical therapeutic targets for eliminating resistant cell pools.

The identified key pathways, Wnt, TGFβ, BMP, and the specific ligands EREG and SEMA3C, along with the metabolic vulnerabilities of the senescent compartment, provide a robust framework for future therapeutic exploration.

Cervical cancer remains a significant global health burden, with chemoradioresistance representing a major obstacle to successful treatment. To elucidate the mechanisms underlying this resistance, we established a unique pair of isogenic primary cervical cancer cell lines, AdMer35 and AdMer43, obtained from a patient with squamous cell carcinoma of the cervix before and after radiation therapy. The aim of our study was to characterize the transcriptomic and cellular heterogeneity of these cells. We conducted an in-depth comparative analysis using single-cell RNA sequencing. Analysis of this paired, patient-derived isogenic model suggests that chemoradioresistance can arise through coordinated multilevel cellular adaptations. Resistant AdMer43 cells demonstrated transcriptional reprogramming, with the upregulation of embryonic stemness factors (HOX, POU5F1, SOX2), a shift in extracellular matrix from fibrillar to non-fibrillar collagens, and activation of inflammatory pathways. We identified and characterized critical cell-state dynamics: resistant cells exhibited a remodeled ecosystem with a metabolically reprogrammed senescent-like cell population showing an enhanced pro-tumorigenic communication via EREG, SEMA3C, BMP, and WNT pathways. Furthermore, we identified a progenitor-like cell population with a minimal CNV burden, potentially serving as a reservoir for tumor persistence. These findings offer novel insights for developing targeted strategies to eliminate resistant cell pools and improve cervical cancer outcomes.