Establishment of Adenomyosis Organoids as a Preclinical Model to Study Infertility

Abstract

Adenomyosis is related to infertility and miscarriages, but so far there are no robust in vitro models that reproduce its pathological features to study the molecular mechanisms involved in this disease. Endometrial organoids are in vitro 3D models that recapitulate the native microenvironment and reproduce tissue characteristics that would allow the study of adenomyosis pathogenesis and related infertility disorders. In our study, human endometrial biopsies from adenomyosis (n = 6) and healthy women (n = 6) were recruited. Organoids were established and hormonally differentiated to recapitulate midsecretory and gestational endometrial phases. Physiological and pathological characteristics were evaluated by immunohistochemistry, immunofluorescence, qRT-PCR, and ELISA. Secretory and gestational organoids recapitulated in vivo glandular epithelial phenotype (pan-cytokeratin, Muc-1, PAS, Laminin, and Ki67) and secretory and gestational features (α-tubulin, SOX9, SPP1, PAEP, LIF, and 17βHSD2 expression and SPP1 secretion). Adenomyosis organoids showed higher expression of TGF-β2 and SMAD3 and increased gene expression of SPP1, PAEP, LIF, and 17βHSD2 compared with control organoids. Our results demonstrate that organoids derived from endometria of adenomyosis patients and differentiated to secretory and gestational phases recapitulate native endometrial-tissue-specific features and disease-specific traits. Adenomyosis-derived organoids are a promising in vitro preclinical model to study impaired implantation and pregnancy disorders in adenomyosis and enable personalized drug screening.

Impaired fertility in adenomyosis: a murine model reveals endometrial receptivity and progesterone resistance imbalances

“…A study on adenomyosis-derived organoids showed the same trend as that in our adenomyosis mice, i.e. increased Lif expression ( Juárez-Barber et al 2022 ). The authors hypothesized that Lif could play a role not only in the implantation process but also in epithelial–mesenchymal transition through the regulation of the Wnt/b-catenin pathway ( Rosario & Stewart 2016 , Juárez-Barber et al 2022 ).…”

“…increased Lif expression ( Juárez-Barber et al 2022 ). The authors hypothesized that Lif could play a role not only in the implantation process but also in epithelial–mesenchymal transition through the regulation of the Wnt/b-catenin pathway ( Rosario & Stewart 2016 , Juárez-Barber et al 2022 ). These findings are crucial for understanding the various mechanisms involved in the physiopathology of adenomyosis and associated symptoms such as infertility and thus finding suitable therapeutic regimens.…”

Macrophage polarization in adenomyosis: A review

“…This technique has been widely used in many fields, such as drug screening and individualized medical treatment 107 . Recently, the AM organoid model has been successfully established, maintaining the biological and pathological characteristics observed in the secretory and gestational eutopic endometria of patients with AM 108 . Therefore, this can be considered for future analysis of AAM polarization and regulatory mechanisms at the 3D organoid level.…”

Hybrid Endometrial‐Derived Hydrogels: Human Organoid Culture Models and In Vivo Perspectives

“…[ 48 ] As with our study, previous works have performed ELISA to characterize the differences between the SPP1 secretion from differentiated and undifferentiated hEOs. [ 6,67 ] The significant upregulation of endometrial receptivity biomarkers ( SPP1, PAEP, LIF, 17HSDβ2 ) [ 52–56 ] together with the repression of the progenitor cell marker SOX9 [ 6,55,56 ] confirmed a higher efficiency of the hEO differentiation in the hybrid endometrial‐derived hydrogel. Altogether, the EndoECM hydrogel component of the hybrid endometrial‐derived hydrogel provides a more suitable microenvironment for endometrial cells, with a matrisome that enhances the ability to differentiate to secretory and gestational phases, and superior rheological properties which can influence signaling pathways that regulate gene expression and cell differentiation.…”