Enrichment of cell cycle pathways in progesterone-treated endometrial organoids of infertile women compared to fertile women

Bich Ngoc Bui, Arif Ibrahim Ardisasmita, F. H. van de Vliert, Matthijs S. Abendroth, Marliek van Hoesel, Shari Mackens, Sabine A. Fuchs, Edward E. S. Nieuwenhuis, Frank J. Broekmans, Gaby S Steba
In: Journal of Assisted Reproduction and Genetics · 2024 · vol. 41(9) , pp. 2405–2418 · doi:10.1007/s10815-024-03173-y · PMID:38995509 · PMC11405558 · W4400583385
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⚙ AI-generated summary by claude@2026-06, 2026-06-10 ⓘ

Progesterone-treated infertile endometrial organoids exhibit enrichment of cell cycle pathways compared to fertile organoids, despite similar proliferation rates.

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⚙ AI-generated deep summary by claude@2026-06, 2026-06-10 ⓘ

This study compared the transcriptome of progesterone-treated endometrial organoids derived from epithelial cells from 14 infertile and seven fertile women, after sequential E2 and progesterone exposure to mimic the secretory phase. Using RNA sequencing with DESeq2, PCA, and functional enrichment analyses (overrepresentation and GSEA), the authors found 363 differentially expressed genes between groups, with cell cycle pathways notably enriched in organoids from infertile women, suggesting potentially higher proliferative activity during differentiation. They also measured proliferation indirectly using OrganoSeg surface metrics before and after differentiation (T1/T0), but this proliferation hypothesis could not be confirmed by the surface measurements. This paper is centrally about endometriosis and/or adenomyosis? It does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Abstract Purpose To investigate whether the transcriptome profile differs between progesterone-treated infertile and fertile endometrial organoids. Methods Endometrial biopsies were obtained from 14 infertile and seven fertile women, after which organoids were generated from isolated epithelial cells. To mimic the secretory phase, organoids were sequentially treated with 17β-estradiol (E2) and progesterone (P4) and subjected to RNA sequencing. Differentially expressed genes (DEGs) were identified using DESeq2 (lfcThreshold = 0, log 2 Fold Change ≥ 1.0 or ≤ −1.0), and a principal component analysis (PCA) plot was generated. Functional enrichment analysis was performed by overrepresentation analysis and Gene Set Enrichment Analysis (GSEA). To functionally assess proliferation, OrganoSeg surface measurements were performed before (T 0 ) and after (T 1 ) differentiation of organoids, and T 1 /T 0 ratios were calculated to determine the proliferation rate. Results Although the PCA plot did not show clear clustering of the fertile and infertile samples, 363 significant DEGs (129 upregulated and 234 downregulated) were detected in infertile compared to fertile organoids. Mainly cell cycle processes were highly enriched in infertile organoids. Thus, we hypothesised that proliferative activity during differentiation may be higher in infertile organoids compared to fertile organoids. However, this could not be validated by cell surface measurements. Conclusions This study revealed that cell cycle processes were enriched in E2/P4-treated infertile endometrial organoids as compared to fertile organoids. This could reflect persistently higher proliferative activity of the endometrial epithelial cells in differentiated infertile organoids compared to fertile organoids. To confirm this hypothesis, further studies are warranted.

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