Exploration of histone protein γ-H2AX as a prognostic factor in soft tissue sarcomas and its association with biological behavior, immune cell environment and survival in leiomyosarcoma

Adrian Georg Simon; Su Ir Lyu; Anne Maria Schultheis; David Stahl; Nora Wuerdemann; Sebastian Walter; Lena Hieggelke; Reinhard Buettner; Christiane Josephine Bruns; Peer Eysel; Lars Mortimer Schiffmann; Karl Knipper; Peter Mallmann; Alexander Quaas; Roland Ullrich

doi:10.1002/ijc.35310

Exploration of histone protein γ-H2AX as a prognostic factor in soft tissue sarcomas and its association with biological behavior, immune cell environment and survival in leiomyosarcoma

Int J Cancer. 2024 Dec 20. doi: 10.1002/ijc.35310. Online ahead of print.

Authors

Adrian Georg Simon¹, Su Ir Lyu¹, Anne Maria Schultheis^{1

2}, David Stahl³, Nora Wuerdemann³, Sebastian Walter⁴, Lena Hieggelke¹, Reinhard Buettner¹, Christiane Josephine Bruns⁵, Peer Eysel⁴, Lars Mortimer Schiffmann⁵, Karl Knipper⁵, Peter Mallmann⁶, Alexander Quaas¹, Roland Ullrich^{3

7}

Affiliations

¹ Institute of Pathology, University Hospital Cologne, Faculty of Medicine, University of Cologne, Cologne, Germany.
² Institute for Surgical Pathology, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
³ Department I of Internal Medicine/Centre for Integrated Oncology Aachen Bonn Cologne Duesseldorf, University Hospital Cologne, Faculty of Medicine, University of Cologne, Cologne, Germany.
⁴ Department for Orthopedics and Trauma Surgery, University Hospital Cologne, Faculty of Medicine, University of Cologne, Cologne, Germany.
⁵ Department of General, Visceral and Cancer and Transplant Surgery, University Hospital of Cologne, Faculty of Medicine, University of Cologne, Cologne, Germany.
⁶ Department of Obstetrics and Gynecology, University Hospital of Cologne, Faculty of Medicine, University of Cologne, Cologne, Germany.
⁷ Centre for Molecular Medicine, University of Cologne, Cologne, Germany.

PMID: 39707602
DOI: 10.1002/ijc.35310

Abstract

This study evaluates the H2AX/γ-H2AX expression in soft tissue sarcomas (STS), its implications for biological behavior and immune environment, and its potential as a prognostic biomarker. RNA-Seq data from 237 STS were obtained from The Cancer Genome Atlas project. Patients were stratified by H2AX mRNA expression using a survival-associated cutoff. Differentially expressed genes and pathways as well as immune signatures between H2AX^high- and H2AX^low tumors were identified with DESeq2 analysis, gene set enrichment analyses (GSEA), Enrichr pathway analysis and CIBERSORTx. Tissue microarrays of a different cohort of 291 STS were generated for immunohistochemical staining to assess γ-H2AX protein expression, followed by statistical evaluation. High H2AX mRNA expression was associated with shorter overall survival (OS) in STS (p = 0.02), particularly in leiomyosarcomas (LMS) (p < 0.001), and was a negative prognostic factor in LMS (HR 11.15, p < 0.001). H2AX^high LMS tumors showed upregulation of cell cycle-related pathways, while H2AX^low LMS exhibited increased inflammatory activity, including elevated M1 macrophage signatures and resting mast cell signatures (both p < 0.001). High γ-H2AX protein levels were an independent negative prognostic factor in the total LMS cohort (HR 12.12, p = 0.025) and in the subgroup of non-uterine LMS (HR 153.80, p = 0.013). Consistent with CIBERSORTx analysis, γ-H2AX^low LMS showed higher mast cell infiltration than γ-H2AX^high LMS (p = 0.038). In conclusion, H2AX mRNA and γ-H2AX protein expression are associated with distinct biological behavior, differences in the immune cell environment, and might serve as useful prognostic biomarkers in LMS.

Keywords: H2AX; biomarker; leiomyosarcoma; soft tissue sarcoma; uterine leiomyosarcoma.