CHD4 R975H mutant activates tumorigenic pathways and promotes stemness and M2-like macrophage polarization in endometrial cancer

Sep 21, 2026

Uploaded by Oliver

Endometrial cancer (EC) remains a disease with poor outcomes and limited effective therapies, making molecular mechanism discovery urgent. A CHD4 R975H mutation was identified as a frequent EC alteration using surveys of EC databases, with computational analyses indicating it disrupts CHD4 structural and functional integrity. Functional experiments show enhanced EC invasion, proliferation, and colony formation, yielding a cancer stem cell-like phenotype. RNA-seq and validation assays demonstrate activation of multiple oncogenic pathways, including NF-κB/TNF-α, KRAS, P53, mTOR, TGF-β, EGFR, Myc, and growth-factor signaling. The mutation also induces stemness and drives tumor-associated macrophages toward an M2-like immunosuppressive state, supporting a dual oncogenic role that informs targeted intervention strategies.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages13

Reading Time33 min

AccessFree to download

Introduction

Endometrial cancer burden and therapeutic need

Cancer stemness and tumor microenvironment

Key genetic alterations in EC

CHD4 background and prior evidence

Methods and Results (from provided content)

Mutation identification and computational prediction

Functional assays for invasion and proliferation

RNA-seq pathway activation and validation

Stemness induction and TAM M2-like polarization

Conclusions and Implications

[www. nature.com/scientificreports](www. nature.com/scientificreports) OPEN CHD4 R975H mutant activates tumorigenic pathways and promotes stemness and M2‑like macrophage polarization in endometrial cancer QinglinZhang1, Fengzhi Zhu2, Yin Tong3, Diwen Shi1 & Jiangwen Zhang1* Endometrial cancer (EC), one of the most prevalent carcinomas in females, is associated with increasing mortality. We identified the CHD4 R975H mutation as a high‑frequency occurrence in EC patients through a comprehensive survey of EC databases. Computational predictions suggest that this mutation profoundly impacts the structural and functional integrity ofCHD4 . Functional assays revealed that the CHD4 R975H mutation enhances EC cell invasion, proliferation, and colony formation, promoting a cancer stem cell (CSC)‑like phenotype. RNA‑seq analysis of cells expressing CHD4 R975H mutant revealed a transcriptomic landscape marked by the activation of several cancer‑promoting signaling pathways, including TNF‑α signaling via NF‑κB, KRAS, P53, mTOR, TGF‑β, EGFR, Myc and growth factor signaling. Validation assays confirmed the activation of these pathways, further demonstrating thatCHD4 R975H mutation induces stemness in EC cells and M2‑like polarization of tumor‑associated macrophages (TAMs). Our study elucidated the oncogenic role ofCHD4 R975H mutation, highlighting its dual impact on facilitating cancer stemness and transforming TAMs into an immunosuppressive subtype. These findings contribute valuable insights into the molecular mechanisms driving EC progression and open avenues for targeted therapeutic interventions. Endometrial cancer (EC), which is derived from endometrium, the inner layer of the uterus, has been regarded as a growing health concern with increasing incidence and cancer-related mortality rates. It is the sixth most prevalent cancer in females, with 417,367 cases diagnosed globally in 2020. Patients in the advanced stages of EC have grim prognoses and increased mortality rates1. Currently, effective therapies for EC are elusive, underscoring the urgency of obtaining a deeper understanding of the molecular mechanisms driving its occurrence and progression. Cancer stemness, delineated by the capacity for self-renewal and initiation of carcinomas by cancer stem cells (CSCs), encompasses a spectrum of cancerous traits propelled by CSC-associated signaling pathways. Its implications include cancer proliferation, invasion, and metastasis2,3. In tumor microenvironment (TME), tumor-associated macrophages (TAMs) predominantly exhibit an immunosuppressive M2-like phenotype, also contributing to the promotion of tumor cell growth, invasion and metastasis4. Previous investigations have underscored the reciprocal relationship between CSCs and TAMs, wherein CSCs have been shown to stimulate M2 polarization of TAMs. This interaction between TAMs and cancer cells not only fosters an immunosuppressive microenvironment but also plays a pivotal role in driving carcinogenesis5,6 Consequently, unravelling the intricacies of the regulatory mechanisms governing cancer stemness and TAMs M2-like polarization in EC patients has profound implications for the development of clinically effective treatments targeting CSCs. Although the molecular intricacies of EC development remain largely elusive, a recent review revealed frequent mutations in a set of tumor-related genes, including PTEN (64–80% in G1–G3 endometrioid endometrial cancer (EEC)), TP53 (59–93% in serous endometrial cancer (SEC)), PIK3CA (22–59% in G1–G3 EEC), and KRAS (19–43% in G1–G3 EEC), among others7. Notably, PTEN has phosphatase activity and functions as a tumor suppressor by inhibiting PI3K-AKT pathway. It converts phosphatidylinositol-3,4,5-trisphosphate 1School of Biological Sciences, The University of Hong Kong, Hong Kong SAR, China. 2College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China. 3Centre for Oncology and Immunology, Hong Kong Science Par

FAQs

How was the CHD4 R975H mutation linked to endometrial cancer in this study?

The mutation was identified as a high-frequency event in EC patients using comprehensive EC database surveys. Computational predictions suggested it significantly affects CHD4 structural and functional integrity.

What functional effects did CHD4 R975H have on endometrial cancer cells?

Functional assays showed increased EC cell invasion, proliferation, and colony formation, supporting a cancer stem cell (CSC)-like phenotype.

Which signaling pathways were activated by CHD4 R975H and what were the downstream impacts?

RNA-seq and validation indicated activation of multiple cancer-promoting pathways, including TNF-α/NF-κB, KRAS, P53, mTOR, TGF-β, EGFR, Myc, and growth factor signaling. These changes supported stemness in EC cells and M2-like polarization of tumor-associated macrophages.

Sep 21, 2026

Uploaded by Oliver