Deconstructing the endometrial disease microenvironment using tissue engineered organoid systems.

Abstract

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. Our lab is a leader in three-dimensional (3D) modelling including tissue clearing, high-content imaging, and create mini uterine tissues using organoids and biomaterials that have enabled us to dissect the 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) 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 a reprogramming toward endometriosis-like disease phenotypes that mimic the invasive, architectural, endocrine and inflammatory hallmarks of endometriosis. These results suggest that matrix and tissue stiffness may be involved in disease progression setting a new role for biomechanics in endometrial reproductive health.

Biography

Dr. Juan S. Gnecco is an Assistant Professor in the Department of Biomedical Engineering and an Associate PI in the Woman, Mother + Baby (WoMB) Research Institute at Tufts University. His research vision lies at the interface of tissue engineering and reproductive biology to understand the immune-endocrine mechanisms driving both reproductive physiology and disease pathogenesis. Dr. Gnecco obtained his B.S. in Biotechnology from Rutgers University, and a PhD in Cellular and Molecular Pathology from Vanderbilt University Medical Center (VUMC) under the mentorship of Dr. Kevin Osteen. In his PhD thesis, he developed the first organ-on-chip model of the perivascular endometrium and deployed this microphysiological systems (MPS) to illuminate the inflammatory effects exerted by environmental toxicants on the female reproductive tract. Dr. Gnecco then conducted his post-doctoral training at Massachusetts Institute of Technology (MIT) with Dr. Linda Griffith in the Department of Biological Engineering where he led efforts to transform the human clinical relevance of 3-dimensional (3D) reproductive tract models by defining the interplay between biophysical and biochemical cues on cell behavior. He developed synthetic extracellular matrices and organoid technologies that helped model and interrogate the tissue microenvironment in vitro. In parallel, he deploys 3D imaging approaches to define the tissue structures using tissue clearing and light-sheet imaging approaches. He has been awarded three rounds of funding from Gates Foundation (2018-present) for his work building phenotypic screening models of the female reproductive tract and was recognized as a Rising Star in Engineering and Health by Columbia University. As the principal investigator of the Laboratory of Reproductive Engineering, his lab focuses on interdisciplinary and translational research by implementing engineering to study the microenvironment in the human female reproductive tract.