{"paper_id":"ff474b81-1c0b-488a-9c96-2a6d09ae69f4","body_text":"Abstract\nBackground\nAdenomyosis (AM) is frequently associated with compromised embryo implantation. The leukemia inhibitory factor (LIF)/signal transducer and activator of transcription 3 (STAT3) signaling pathway is crucial for endometrial receptivity; however, its mechanistic role in AM-related implantation failure remains insufficiently elucidated. This study aimed to investigate the inactivation mechanism of the LIF/STAT3 pathway in AM and to evaluate a novel nanotherapeutic strategy for restoring implantation capacity.\nMethods\nClinical endometrial samples from AM patients and controls were analyzed. A three-dimensional (3D) co-culture system simulating the endometrial microenvironment was established. Interventions were performed using a tofacitinib-loaded biomimetic nanogel (Tofa-NG). Comprehensive analyses included bulk RNA Sequencing (RNA-seq), immunohistochemistry (IHC), RT-qPCR, proteomics, inflammatory cytokine profiling, and metabolomics to dissect the pathological links.\nResults\nThe LIF/STAT3 signaling pathway was significantly downregulated in the endometrium of AM patients, correlating directly with implantation failure. Pathologically elevated inflammatory cytokines suppressed LIF expression via NF-κB pathway activation, exacerbating the inflammatory microenvironment. Metabolomic profiling revealed a strong association between LIF/STAT3 pathway inactivation and aberrant cellular energy metabolism. The engineered Tofa-NG facilitated targeted drug delivery, effectively mitigated local inflammation, and successfully reactivated the LIF/STAT3 pathway. Consequently, this intervention significantly improved the embryo implantation success rate in the experimental model.\nConclusion\nThis study identifies the inactivation of the LIF/STAT3 pathway as a central mechanism underlying embryo implantation defects in AM, intricately linked to chronic inflammation and metabolic dysregulation. The Tofa-NG strategy demonstrates promising therapeutic potential by rectifying the signaling deficit and ameliorating the endometrial microenvironment. These findings provide a novel theoretical foundation for developing targeted, personalized treatments for infertility associated with AM.\nGraphical abstract\nSimilar content being viewed by others\nAbbreviations\n- AM:\n-\nAdenomyosis\n- ANOVA:\n-\nAnalysis of variance\n- ATP:\n-\nAdenosine triphosphate\n- BSA:\n-\nBovine serum albumin\n- BP:\n-\nBiological processes\n- CC:\n-\nCellular components\n- CS:\n-\nChitosan\n- DEGs:\n-\nDifferentially expressed genes\n- DEPs:\n-\nDifferentially expressed proteins\n- DHE:\n-\nDihydroethidium\n- ECM:\n-\nExtracellular matrix\n- EECs:\n-\nEndometrial epithelial cells\n- FDR:\n-\nFalse discovery rate\n- GO:\n-\nGene ontology\n- GSEA:\n-\nGene set enrichment analysis\n- HESC:\n-\nHuman endometrial stromal cell\n- H&E:\n-\nHematoxylin and eosin\n- iBAQ:\n-\nIntensity-based absolute quantification\n- IHC:\n-\nImmunohistochemistry\n- KEGG:\n-\nKyoto encyclopedia of genes and genomes\n- LC-MS/MS:\n-\nLiquid chromatography-tandem mass spectrometry\n- LFQ:\n-\nLabel-free quantification\n- LIF:\n-\nLeukemia inhibitory factor\n- m6A:\n-\nN6-Methyladenosine\n- MeRIP-qPCR:\n-\nMethylated RNA immunoprecipitation followed by quantitative polymerase chain reaction\n- MF:\n-\nMolecular functions\n- NES:\n-\nNormalized enrichment scores\n- NGF:\n-\nNerve growth factor\n- PAA:\n-\nPolyacrylic acid\n- PCA:\n-\nPrincipal component analysis\n- PBS:\n-\nPhosphate-buffered saline\n- PLS-DA:\n-\nPartial least squares discriminant analysis\n- RIP-qPCR:\n-\nRNA immunoprecipitation followed by quantitative polymerase chain reaction\n- RNA-seq:\n-\nRNA sequencing\n- ROS:\n-\nReactive oxygen species\n- SMPDB:\n-\nSmall molecule pathway database\n- STAT3:\n-\nSignal transducer and activator of transcription 3\n- TLR4:\n-\nToll-like receptor 4\n- Tofa:\n-\nTofacitinib\n- Tofa-NG:\n-\nTofacitinib-loaded biomimetic nanogel\n- VAS:\n-\nVisual analogue scale\n- VIP:\n-\nVariable importance in projection\n- WB:\n-\nWestern blot\n- Δψm:\n-\nMitochondrial membrane potential\n- 2D:\n-\nTwo-dimensional\n- 3D:\n-\nThree-dimensional\nAcknowledgements\nNone.\nFunding\nFunded by Science and Technology Program of Xuzhou (Grant No. KC25074), and Science and Technology Project of Jiangsu Provincial Administration of Traditional Chinese Medicine (Grant No. MS2023079), and “Jinding” Reserve Talents Project of the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University (Grant No. XYYJDYCHB202501).\nAuthor information\nAuthors and Affiliations\nCorresponding author\nEthics declarations\nEthics approval and consent to participate\n1. This study was approved by the Medical Ethics Committee of the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University (Approval No. Xyyll [2025] 167). All patients included in the study provided informed consent. 2. All the animal experiments were approved by the Animal Experiment Ethics Committee of the Jiangsu Provincial Hospital of Integrated Traditional Chinese and Western Medicine (AEWC-20241009443).\nConsent for publication\nAll authors agree to publish the article.\nCompeting interests\nThe authors declare 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\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\nDou, Q., Han, Z., Shao, W. et al. Inactivation of the leukemia inhibitory factor/signal transducer and activator of transcription 3 pathway: a pivotal mechanism in adenomyosis-related embryo implantation failure. J Nanobiotechnol (2026). https://doi.org/10.1186/s12951-026-04505-z\nReceived:\nAccepted:\nPublished:\nDOI: https://doi.org/10.1186/s12951-026-04505-z","source_license":"public-domain-us","license_restricted":false}