Endometriosis is a chronic inflammatory disease affecting over 190 million women worldwide and is characterized by the growth of endometrial-like tissue outside the uterine cavity. Dysregulation of endocrine signals play an important role in the pathogenesis of endometriosis and influences symptoms like severe, chronic pelvic pain; however, the underlying mechanisms of disease pathogenesis remain elusive. A better understanding of the cellular and biomechanical components of the lesion microenvironment is needed to understand the early events in disease establishment. Recent technological advances in three-dimensional (3D) modelling including tissue clearing, high-content imaging, organoids and biomaterials that have enabled our team to dissect and reconstruct the endometriotic tissue microenvironment. Using patient-derived biospecimens we describe the tools that have transformed our understanding of origins of endometriosis. We deploy tissue clearing methods (i.e., CLARITY) and 3D imaging of human endometriotic (adenomyosis and endometriosis, N=10) lesions to spatially characterize the lesion architecture and extracellular matrix (ECM) composition. From these results, we refined the hypothesis that lesion morphology is linked to bulk tissue mechanics. To test this, we used our recently developed functionalized poly-ethylene glycol (PEG) synthetic matrices and multi-cellular organoid systems to explore tissue biomechanics and cell-cell communication in endometriotic disease. By embedding endometrial epithelial organoids (EEOs) in PEG matrices tuned to mimic physiologic (3wt%, 300pa) to pathologic (5wt%, 2kPa) regimes we identified morphometric, mitotic and inflammatory profiles using immunofluorescent imaging, transcriptomics and multiplexed bead arrays to identify the reprogramming toward an endometriotic-like disease phenotype that mimic the invasive, architectural, endocrine and inflammatory hallmarks of endometriosis. These results suggest that matrix and tissue biomechanics may be involved in disease progression setting a new role for biomechanics in endometrial reproductive health.
