Towards a Non-Invasive Model for Women’s Uterine Health: From Transcriptional Programs to Tissue Assembly

Abstract

In human females of reproductive age, the uterine endometrium undergoes a cyclic process of hormone-induced rapid cellular differentiation, enhanced vascular development, matrix remodeling, and structural change. Functional reproduction of such a dynamic organ system in vitro requires the engineering of hormonally responsive cells, niche-like structures, and engineered vasculature to ideally recapitulate the uterine endometrium in both the proliferative and secretory phase. This thesis is focused on developing novel technologies to engineer biomimetic in vitro tissue models using induced pluripotent stem cells (hiPSCs), as well as bioinformatic analysis on the origin of the uterine-related disease endometriosis. Chapter 1 describes the engineering of endometrial stromal and glandular-like cells from pluripotent stem cells using combinatorial transcription factor expression. We also demonstrate the creation of niche-like extracellular matrix (ECM) hydrogel wells for in vitro culture of murine and synthetic murine embryos. Finally, we combine the techniques and embed candidate murine endometrial-like tissues in engineered hydrogel niches and co-culture the synthetic endometrial tissue with murine embryos. Chapter 2 details the acquisition, processing, and sequencing of the whole transcriptome of a clinical endometriosis patient cohort with a specific emphasis on distinguishing lesions by color phenotype. We found color-specific markers and oncogenes which support a color-specific etiology and therapy modalities. Chapter 3 describes the generation of a brain organoid model. We utilized transcription factor overexpression to create mature endothelium and neurons within days and embedded them in a layered architecture, creating a vascularized brain organoid model with an inner core of neural stem cells, a layer of induced mature neurons, and a pervasive vascular network.