Eric Parigoris
BME PhD Defense Presentation

Date:2022-05-09
Time: 10:00 AM - 12:00 PM ET
Location / Meeting Link: EBB CHOA Room or https://teams.microsoft.com/l/meetup-join/19%3ameeting_N2NlYzU0MWMtN2MzZC00NjEzLThmNzUtYmMzNzYzOTU3MWUz%40thread.v2/0?context=%7b%22Tid%22%3a%22482198bb-ae7b-4b25-8b7a-6d7f32faa083%22%2c%22Oid%22%3a%228944e5f6-f256-45fa-ab5c-74a0be29407a%22%7d

Committee Members:
Shuichi Takayama, Ph.D. (Advisor) YongTae Kim, Ph.D. David Myers, Ph.D. Todd Sulchek, Ph.D. Xueying Zhao, Ph.D


Title: Geometrically-Inverted Organoids

Abstract: To minimize the cost, ethics, and species-specific variations associated with animal models, in vitro models are gradually becoming adopted, as they have improved our understanding of both normal and diseased organ function. This thesis focuses on organoids, which are 3-dimensional human cell cultures that mimic the physiological architecture of native organs, and exhibit advantages over other in vitro models such as the ability to be cultured long-term and grown in high-throughput formats. However, one of the main limitations of organoid models is the inability to access the apical side, as it faces the interior of traditional organoids (apical-in, basal-out). Recently, researchers have circumvented this issue by engineering “geometrically-inverted” organoids, in which the apical (apical-out) and basolateral (basal-in) poles are reversed. Such an advancement opens the possibility for new applications which require apical access, including cancer invasion, nutrient uptake, and viral infection studies. While there have been a few studies that establish these inverted organoid models, some challenges remain such as the need to manipulate the extracellular matrix environment to cause eversion, the need for large Matrigel amounts, and the difficult organoid removal process from Matrigel. This thesis therefore aims to establish a geometrically-inverted organoid platform that address these challenges by engineering stably inverted and free-floating organoids that require low Matrigel amounts, and that are compatible with high-throughput microscopy. Aim 1 will establish the platform for geometrically-inverted organoids using a breast model, and show its applicability in a physiologically consistent cancer invasion assay. Aim 2 will expand this mammary organoid model to include co-cultures with adipocytes, and further establish a high-throughput imaging workflow for the organoids. Finally, Aim 3 will engineer and characterize the first reported geometrically-inverted proximal tubule organoid and use it to study proteinuria (high protein levels). While this thesis focuses on inverted breast and kidney organoids, the methods and concepts may be applicable to a broad range of organ, tissue, and cancer types.