RNA-seq and ATAC-seq analysis of CD163⁺ macrophage-induced progestin-insensitive endometrial cancer cells

Progestins are used as fertility-sparing regimens for young patients with stage 1A endometrioid endometrial cancer (EEC) and atypical endometrial hyperplasia (AEH). CD163⁺ macrophages promote estrogen-dependent EEC development, but whether they induce progestin insensitivity remains unclear. This study aimed to investigate the possible effects of CD163⁺ macrophages on progestin response in AEH/EEC patients.

The number of infiltrating CD163⁺ macrophages in progestin-insensitive and -sensitive endometrial lesions was compared. The effects of CD163⁺ macrophages on progestin responses and progesterone receptor (PR) expression in EC cells were evaluated in vitro. ATAC-seq and RNA-seq were combined to identify molecular/biological changes induced by CD163⁺ macrophages in progestin-insensitive EC cells.

Increased CD163⁺ macrophage infiltration was significantly associated with progestin insensitivity and longer treatment durations in AEH/EEC patients. Additionally, the number of CD163⁺ macrophages was negatively correlated with PR expression in AEH/EEC tissues. Furthermore, the CD163⁺ macrophage-mediated microenvironment and secreted cytokines downregulated PR expression and impaired the response of EC cells to medroxyprogesterone acetate (MPA). RNA-seq analysis demonstrated that CD163⁺ macrophages antagonized PR signaling by blocking or even reversing MPA-regulated differential gene expression. Based on RNA-seq and ATAC-seq analyses, extracellular matrix (ECM) signaling and ECM-related transcription factors, FOXF2, POU1F1, and RUNX1were identified to potentially be involved in CD163⁺ macrophage-induced progestin insensitivity in endometrial cancer patients.

We identified CD163⁺ macrophages as an important mediator of progestin desensitization and an unfavorable factor for the efficacy of fertility-preserving treatment in AEH/EEC patients.