Dear Office of Graduate studies,

Below are the details for my Phd defense:

Title: Human Airway Organoids with Reversed Biopolarity: Implications for Infectious Disease, Drug Discovery, and Pathophysiology

Date: 2024-04-12
Time: 1:00 pm
Location: EBB CHOA Room
Meeting Link: https://gatech.zoom.us/j/6292679646?pwd=THZxSkFSUExuekZIN0RLalh4clVMZz09

Advisor: Shuichi Takayama, PhD (Georgia Institute of Technology)

Committee Members:
Stanislav Emelianov, PhD (Georgia Institute of Technology)

M. G. Finn, PhD (Georgia Institute of Technology)

Eric J. Sorscher, MD (Emory University School of Medicine)

Jason R. Spence, PhD (University of Michigan)

Abstract:

Organoid research is exploring a new niche: apical-out organoids. These engineered structures, characterised by their outward-facing apical membrane, offer unique and exciting avenues for research. This dissertation introduces human airway organoids with reversed polarity (AORBs) and demonstrates their potential in studying infectious diseases, drug discovery, and pathophysiology, with a specific focus on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and bronchiolitis obliterans syndrome (BOS). First, we successfully established a robust and optimised standard operating procedure (SOP) for generating AORBs from human primary upper airway epithelial cells, building upon a previously described minimal extracellular matrix (ECM) scaffolding method. The resulting AORBs were cultured free-floating in a high-throughput, single-organoid-per-well format. Single-cell RNA sequencing confirmed the in vivo-like cellular heterogeneity of AORBs, validating their close resemblance to natural airway physiology. Next, we demonstrated the utility of AORBs as a platform for SARS-CoV-2 infection and antiviral drug screening. AORBs supported efficient infection by five SARS-CoV-2 variants, while enabling high-throughput screening, effectively identifying false negatives and false positives. Furthermore, the SOP was refined to develop a tri-culture organoid model, integrating airway epithelial cells, fibroblasts, and peripheral blood stem cells, with the objective of addressing the lack of a human model and the limited translatability of existing animal models in studying BOS pathogenesis. By manipulating human leukocyte antigen matching between epithelial and immune cells in vitro, our BOS model can closely recapitulate the clinical condition, facilitating mechanistic studies and therapeutic discovery. Overall, this dissertation establishes AORBs as a versatile and standardised platform for investigating infectious diseases, drug discovery, and pathophysiology of the respiratory system. The standardisation capacity of the developed SOP holds broader implications for standardising organoids, aligning with the recently updated FDA regulation lifting the mandate for animal studies on therapeutic testing.