Illuminating epithelial-stromal communication using engineered synthetic matrix microenvironments

Abstract

Mucosal barrier tissues are prominent targets for drugs against infection and chronic inflammatory disorders. One such mucosal barrier tissue, the endometrium, undergoes monthly cyclic remodeling via hormone-mediated growth, immune cell recruitment and proteolytic breakdown. Hormone response disruption has been associated with numerous endometrial pathologies, including endometriosis, adenomyosis, and infertility, which impacts upwards of 10% of women during their reproductive years. Currently, our understanding of endometrial biology is limited by the ability to replicate complex 3D physiology in vitro. Our ability to parse disease mechanisms and test efficacy of therapeutic interventions relies on development of reproducible models, adaptable to the limited numbers of cells available from patient biopsies. In this thesis, I address a critical gap in accessible tools to study and control endometrial biology in vitro and do so in a manner that can be translated to other epithelial-stromal mucosal tissues. Using the endometrium as an example mucosal barrier, I first establish design principles for the development of a synthetic, modular extracellular matrix (ECM) hydrogel suitable for 3D functional co-culture of epithelial and stromal cells. This 'one-size- fits-all' matrix features components that can be remodeled by cells and that responds dynamically to sequester local cell-secreted ECM characteristic of each cell type enabling long-term, hormonally responsive co-cultures. Next, I establish methods to expand and cryopreserve primary human endometrial epithelial cells, which maintain barrier and secretory function, further enabling studies using primary cells. Finally, we use data-driven network modeling of secreted proteins to understand how variation in cytokine signaling may alter hormone responsiveness and proteolytic remodeling in primary epithelial-stromal co-cultures. With the ability to create and parse more complex 3D tissue models using primary cells to recapitulate healthy and diseased states, we further enable basic understandings of disease pathologies and subsequent drug discovery efforts aimed at inflammation, wound healing and immune modulation.