Individual-dependent molecular heterogeneity in human endometrial stromal cells: a potential determinant of female fertility

Background Human endometrial stromal cells (hEnSCs) are central to female fertility. This study investigated the cellular and molecular landscape of hEnSCs and identified inter-individual variations that may influence reproductive potential.

and results Endometrial biopsies were obtained from 16 women undergoing gynecologic evaluation. Primary hEnSCs were isolated, expanded, and characterized for mesenchymal stem cell properties. Stemness, receptivity, and decidualization capacity were assessed using RT-qPCR and RNA sequencing, followed by differential gene expression and pathway enrichment analyses. Integrated cellular and transcriptomic analyses revealed donor-dependent molecular heterogeneity. Key genes involved in stemness (OCT4, KLF4, MYC), decidualization (PRL, IGFBP1), and endometrial receptivity (ESR1, PGR, HOXA10, and HOXA11) showed significant variability among individuals. RNA-seq analyses further identified differentially expressed genes associated with clinical variables, including age, body mass index (BMI), and endometrial thickness. Age-related transcriptomic changes included enrichment of genes involved in extracellular matrix organization, cell adhesion, and immune regulation. A higher BMI was connected to altered regulation of genes related to the cell cycle, extracellular matrix, and signaling receptor regulatory activity, while a thin endometrium was associated with downregulation of genes involved in cell-cell junction, signaling receptor regulatory activity, and epidermal development.

hEnSCs exhibit significant individual-dependent molecular heterogeneity, with donor-specific variations influencing crucial biological pathways related to receptivity, stemness, and decidualization. These molecular profiles, influenced by age, BMI, and endometrial thickness, may reflect the fertility potential of the endometrium. Recognition of such intrinsic, individual-specific endometrial variation emphasizes the importance of personalized approaches in fertility assessment and reproductive treatments. Similar content being viewed by others Data availability Data will be made available to the editors of the journal for review or query upon request to the corresponding author. Abbreviations - OCT4: - POU class 5 homeobox 1(POU5F1) - KLF4: - Kruppel like factor 4 - SOX2: - SRY-box transcription factor 2 - MYC: - MYC proto-oncogene, bHLH transcription factor - ESR 1: - Estrogen receptor 1 - PGR: - Progesterone receptor - HOXA10: - Homeobox A10 - HOXA11: - Homeobox A11 - PRL: - Prolactin - IGFBP1: - Insulin-like growth factor binding protein 1 - LEP: - Leptin - FABP4: - Fatty Acid Binding Protein 4 - COL1A1: - Collagen Type I Alpha 1 - SPARC: - Osteonectin - ACAN: - Aggrecan - COL2A1: - Collagen Type II Alpha 1 - GAPDH: - Glyceraldehyde-3-phosphate dehydrogenase - LIF: - Leukemia inhibitory factor - MMP7: - Matrix metallopeptidase 7 - MUC1: - Mucin 1, cell surface associated - ITGAV: - Integrin subunit alpha V - ANXA2: - Annexin A2 - IL15: - Interleukin 15 - IGF1R: - Insulin-like growth factor 1 receptor - BMP2: - Bone morphogenetic protein 2 - DKK1: - Dickkopf WNT signaling pathway inhibitor 1 - ITGA4: - Integrin subunit alpha 4 - DCN: - Decorin - HAND2: - Heart and neural crest derivatives expressed 2 - ELN: - Elastin - TIMP3: - TIMP metallopeptidase inhibitor 3 - BMI1: - BMI1 proto-oncogene, polycomb ring finger - SALL4: - Spalt like transcription factor 4 - KLF2: - Kruppel-like factor 2 - PPIA: - Peptidylprolyl isomerase A (Cyclophilin A) - ALDH1A1: - Aldehyde dehydrogenase 1 family member A1 - SOX15: - SRY-box transcription factor 15 - NOTCH1: - Notch receptor 1 - SLC34A2: - Solute carrier family 34 member 2 - HOXB9: - Homeobox B9 - HSH2D: - Hematopoietic SH2 domain containing - RASSF6: - Ras association domain family member 6 - DOC2B: - Double C2 domain beta - NMUR2: - Neuromedin U receptor 2 - VAV1: - Vav guanine nucleotide exchange factor 1 - MUC5AC: - Mucin 5AC, oligomeric mucus/gel-forming - PLVAP: - Plasmalemma vesicle associated protein - TDRD15: - Tudor domain containing 15 - CX3CL1: - C-X3-C motif chemokine ligand 1 (Fractalkine) - CNTNAP2: - Contactin associated protein-like 2 - SERPINA1: - Serpin family A member 1 (Alpha-1-antitrypsin) - LCN2: - Lipocalin 2 - MDFI: - MyoD family inhibitor - COL22A1: - Collagen type XXII alpha 1 chain - TDRD15: - Tudor domain containing 15 - SLA: - Src-like adaptor - TNFSF18: - Tumor necrosis factor superfamily member 18 (GITRL) - ACTG2: - Actin gamma 2, smooth muscle - NKX3-2: - NK3 homeobox 2 - CCDC144A: - Coiled-coil domain containing 144 A - EPYC: - Epiphycan - PLXDC1: - Plexin domain containing 1 - C11orf87: - Chromosome 11 open reading frame 87 - GRM4: - Glutamate metabotropic receptor 4 - SPHKAP: - Sphingosine kinase type 1 interacting protein - NKX2-6: - NK2 homeobox 6 - LCE2A: - Late cornified envelope 2 A - HDC: - Histidine decarboxylase - LYPD6B: - LY6/PLAUR domain containing 6B - SBSN: - Suprabasin - PLAC8: - Placenta-specific 8 - MGAM: - Maltase-glucoamylase - PTPRQ: - Protein tyrosine phosphatase receptor type Q - MAB21L1: - Mab-21 like 1 - GLDN: - Gliomedin - ABI3: - ABI family member 3 (Abl interactor 3) - DMRTA2: - DMRT like family A2 - EPHA3: - EPH receptor A3 - HLA-DPA1: - Major histocompatibility complex, class II, DP alpha 1 - HLA-DQB1: - Major histocompatibility complex, class II, DP beta 1 - PWP2: - Periodic tryptophan protein 2 homolog - VIT: - Vitrin - TMEM100: - Transmembrane protein 100 - GPRIN2: - G protein regulated inducer of neurite outgrowth 2 - PRODH: - Proline dehydrogenase 1 - UGT1A6: - UDP glucuronosyltransferase family 1 member A6 - CASC19: - Cancer susceptibility 19 (long non-coding RNA) - ADAMTSL3: - ADAMTS like 3 - LINC01411: - Long intergenic non-protein coding RNA 1411 - SLC52A3: - Solute carrier family 52 member 3 (riboflavin transporter 3) - NOS1: - Nitric oxide synthase 1 - CA2: - Carbonic anhydrase 2

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FASEB J. https://doi.org/10.1096/fj.202101579R Funding This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT (MSIT) of the Government of South Korea (Grant Number: RS-2024-00405287) and grant funded by the Institute of Planning and Evaluation for Technology (IPET) through Ministry of Agriculture, Food and Rural Affairs (MAFRA) (grant number: 321025-05). Author information Authors and Affiliations Contributions Concept/design: U.R., E.Y.K, D.C.M.K., Y.B.K., and M.K.K.Sample collection: U.R., E.Y.K, R.E.K., J.W.H., and Y.J.K.Experiments: U.R., E.Y.K., D.C.M.K., R.E.K., J.W.HData analysis: U.R., and D.C.M.KBioinformatics: U.R., and D.C.M.KManuscript drafting: U.R., E.Y.K., D.C.M.K., and M.K.K.Manuscript revision: U.R., E.Y.K., D.C.M.K., R.E.K., J.W.H., Y.J.K., Y.B.K., and M.K.K.Supervision: M.K.K.Funding acquisition: M.K.K. Corresponding author Ethics declarations Competing interests The authors declare no competing interests. Ethics approval This study was performed in accordance with the principles of the Declaration of Helsinki. Approval was granted by the Institutional Review Board of Chungnam National University Hospital (IRB no. CNUH 2022-11-002). Consent to participate Informed consent was obtained from all individual participants included in the study. Consent to publish Not applicable. Additional information Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Below is the link to the electronic supplementary material. 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