{"paper_id":"ec335170-2964-495e-8645-2a5655d6dbb3","body_text":"Abstract\nBackground\nDespite advancements in imaging techniques, the non-invasive diagnosis of adenomyosis remains particularly challenging due to the overlap of imaging features with other uterine diseases, and the lack of consistent correlation with clinical presentation, with histological confirmation after hysterectomy still necessary. In this context, circulating miRNAs emerge as promising candidates for accurate non-invasive diagnosis.\nMethods\nTo determine whether women with adenomyosis exhibit a distinct circulating plasma miRNA profile that can serve as a diagnostic marker, miRNA sequencing was performed to compare the plasma miRNA profile of women with adenomyosis with infertile and healthy fertile controls (n = 10/group). Adenomyosis was diagnosed by transvaginal ultrasound using Morphological Uterus Sonographic Assessment (MUSA) criteria. Plasma and endometrial biopsies were collected during the secretory phase from women undergoing hormone replacement therapy. Total RNA was extracted from plasma, libraries were prepared using NEXTFLEX® Small RNA-Seq v4 kits, and miRNAseq was performed on the NextSeq 550 NGS platform. Differentially expressed (DE) miRNAs were identified using the edgeR method (false discovery rate < 0.05). The discriminatory potential of DEmiRNAs was evaluated by receiver operating characteristic (ROC) curve analysis using the area under the curve (AUC). Validation was performed by quantitative PCR on plasma and endometrium (n = 15/group). The target genes of DEmiRNAs were retrieved from miRTarBase and subjected to functional enrichment analysis.\nResults\nA total of 11 disease-specific DEmiRNAs were identified in adenomyosis compared to infertile controls (5 up- and 6 downregulated), and 23 healthy control-referenced DEmiRNAs compared to fertile controls (19 up- and 4 downregulated). Among disease-specific DEmiRNAs, miR-4433 and miR-652-5p demonstrated strong diagnostic performance for adenomyosis compared to infertile controls (AUC = 0.92 and 0.91, respectively; p < 0.001). For healthy control-referenced DEmiRNAs, 11 miRNAs showed statistically significant diagnostic value (p < 0.01), with the highest AUCs observed for miR-3200-5p, miR-193b-5p, and miR-4732-5p (AUC = 0.88, 0.88 and 0.87, respectively). Validation experiments corroborated the upregulation of miR-4433 and miR-92a-3p upregulation in plasma and endometrium.\nConclusions\nThis study supports the potential of circulating miRNAs as non-invasive biomarkers for adenomyosis and contributes to understanding its molecular basis. The concordance between circulating and endometrial miRNA expression suggests a possible link between systemic molecular alterations and endometrial pathology.\nSimilar content being viewed by others\nAbbreviations\n- AUC:\n-\nArea under the curve\n- BP:\n-\nBiological process\n- BMI:\n-\nBody mass index\n- CC:\n-\nCellular component\n- DEmiRNAs:\n-\nDifferentially expressed miRNAs\n- EMT:\n-\nEpithelial-to-mesenchymal transition\n- FDR:\n-\nFalse discovery rate\n- GO:\n-\nGene Ontology\n- KEGG:\n-\nKyoto Encyclopedia of Genes and Genomes pathways\n- MRI:\n-\nMagnetic resonance imaging\n- miRNAs:\n-\nMicroRNAs\n- MF:\n-\nMolecular function\n- MUSA:\n-\nMorphological Uterus Sonographic Assessment\n- ORA:\n-\nOver-Representation Analysis\n- ROC:\n-\nReceiver operating characteristic\n- TVUS:\n-\nTransvaginal ultrasound\nAcknowledgements\nWe sincerely thank all the women who kindly agreed to participate in this study. We also acknowledge the IVIRMA Clinic staff for their assistance in sample collection, the Genomics Service of the University of Valencia for their technical assistance with miRNA sequencing, and EpiDisease for their support in data analysis.\nFunding\nThis project was funded by the Conselleria de Educación, Universidades y Empleo (Generalitat Valenciana) (CIGE/2023/72 awarded to A.C.); the Health Institute Carlos III, co-funded by the European Social Fund (ESF) ‘Investing in your future’ (CP20/00120 awarded to H.F.; CD23/00157 awarded to A.C.).\nAuthor information\nAuthors and Affiliations\nCorresponding authors\nEthics declarations\nEthics approval and consent to participate\nThe collection and use of human samples was approved by the Clinical Research Ethics Committee at Hospital La Fe (2301-FIVI-008-HF) and informed written consent was obtained from all participants.\nConsent for publication\nNot applicable.\nCompeting interests\nThe authors declare that they have no competing interests.\nAdditional information\nPublisher’s Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nSupplementary Information\n12958_2026_1598_MOESM1_ESM.xlsx (download XLSX )\nSupplementary Material 1: Supplementary Table 1. Differentially expressed microRNAs in plasma from women with adenomyosis.\n12958_2026_1598_MOESM2_ESM.xlsx (download XLSX )\nSupplementary Material 2: Supplementary Table 2. Predicted target genes of the differentially expressed microRNAs in plasma from women with adenomyosis compared with infertile controls.\n12958_2026_1598_MOESM3_ESM.xlsx (download XLSX )\nSupplementary Material 3: Supplementary Table 3. Genes most commonly targeted by differentially expressed microRNAs in the adenomyosis group compared with the infertile control group.\n12958_2026_1598_MOESM4_ESM.xlsx (download XLSX )\nSupplementary Material 4: Supplementary Table 4. Kyoto Encyclopedia of Genes and Genomes pathway enrichment of predicted target genes of differentially expressed plasma microRNAs in adenomyosis compared with infertile controls.\n12958_2026_1598_MOESM5_ESM.xlsx (download XLSX )\nSupplementary Material 5: Supplementary Table 5. Gene Ontology enrichment analysis of predicted target genes of differentially expressed plasma microRNAs in adenomyosis compared with infertile controls.\n12958_2026_1598_MOESM6_ESM.xlsx (download XLSX )\nSupplementary Material 6: Supplementary Table 6. Predicted target genes of the differentially expressed microRNAs present in plasma from women with adenomyosis compared with fertile controls.\n12958_2026_1598_MOESM7_ESM.xlsx (download XLSX )\nSupplementary Material 7: Supplementary Table 7. Genes most commonly targeted by differentially expressed microRNAs in the adenomyosis group compared with the fertile control group.\n12958_2026_1598_MOESM8_ESM.xlsx (download XLSX )\nSupplementary Material 8: Supplementary Table 8. Kyoto Encyclopedia of Genes and Genomes pathway enrichment of predicted target genes of differentially expressed plasma microRNAs in adenomyosis compared with fertile controls.\n12958_2026_1598_MOESM9_ESM.xlsx (download XLSX )\nSupplementary Material 9: Supplementary Table 9. Gene Ontology enrichment analysis of predicted target genes of differentially expressed plasma microRNAs in adenomyosis compared with fertile controls.\nRights and permissions\nOpen Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.\nAbout this article\nCite this article\nCorachán, A., Pérez-Ferrer, M., Faus, A. et al. Circulating plasma microRNAs as potential non-invasive biomarkers in infertile women with adenomyosis: an observational study. Reprod Biol Endocrinol (2026). https://doi.org/10.1186/s12958-026-01598-x\nReceived:\nAccepted:\nPublished:\nDOI: https://doi.org/10.1186/s12958-026-01598-x","source_license":"CC0","license_restricted":false}