Integrative single-cell sequencing and machine learning analyses identify chronic inflammation and cancer-associated fibroblast subsets as drivers of adverse outcomes in ovarian cancer.

Authors

Mingqin Kuang, Changmei Shen, Chunping Yang, Shuchun Xie, Yiheng Luo, Baozhen Liao, Hailong Chen

Published in

Translational oncology. Volume 72. Pages 102992. Aug 22, 2026. Epub Aug 22, 2026.

Abstract

Ovarian cancer remains the deadliest gynecological malignancy, with neoadjuvant chemotherapy (NACT) often leaving residual fibroblast-enriched disease.
Single-cell RNA sequencing data from 64,097 cells were analyzed using Seurat (v4.1.3) with Harmony batch correction. CellChat (v1.1.3) mapped intercellular communication, while Monocle3 constructed pseudotime trajectories. High-dimensional weighted gene co-expression network analysis (hdWGCNA) identified fibroblast subclusters with soft threshold β=12 (scale-free R²=0.8). Six machine learning algorithms screened hub genes across three bulk RNA-seq datasets. ROC curves evaluated diagnostic performance. In vitro validation included siRNA-mediated NHP2 knockdown in OVCAR-8 and A2780 cells, with proliferation assessed by CCK-8, migration, and invasion by Transwell assays.
Compared to pre-NACT, post-NACT tissues exhibited increased fibroblast and CD8+ T cell proportions, with reduced dendritic cells. CellChat analysis revealed selective communication between epithelial cells, monocytes, CD8+ T cells, and fibroblasts. hdWGCNA identified the M8 fibroblast subcluster as the predominant post-chemotherapy population, showing diffuse spatial distribution and strongest association with tumor epithelial cells. Differential expression analysis identified 15 upregulated genes post-NACT, with NHP2 showing optimal diagnostic value (AUC=0.758). SVM algorithm identified CALU, NHP2, ANXA5, CUTA, and TUBA1B as hub genes. High NHP2 expression correlated with poor overall survival (HR=1.26, 95% CI: 1.10-1.43, P = 0.00,057) and progression-free survival (HR=1.23, 95% CI: 1.08-1.40, P = 0.0018). NHP2 knockdown significantly inhibited proliferation (P < 0.001), invasion (P < 0.01), and migration (P < 0.001).
The M8 fibroblast subcluster and NHP2 represent critical mediators of chemotherapy resistance and immunosuppression in ovarian cancer, offering promising therapeutic targets to improve patient outcomes.

PMID:
42632123
Bibliographic data and abstract were imported from PubMed on 23 Aug 2026.

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