RECODE: Using light and mechanics to monitor and control the differentiation of lung alveolar organoids

Princeton University, Princeton NJ

Investigators

Abstract

Cells can be coaxed into forming organ-like structures outside of the body. These “organoids” would help scientists study organ development, function, and disease. Organoids often fail to form in a reproducible manner, currently limiting their utility. This project will invent new approaches to reliably build organoids that mimic the lung. The investigators will track and control organoid formation using light and mechanical forces. The project will introduce high school and college students from underrepresented communities to scientific research. The project will also share its approaches by building an international symposium. The minimal functional unit of the lung is the alveolus, which is comprised of alveolar epithelial type I (AT1) cells interspersed with type II (AT2) cells, surrounded by a meshwork of myofibroblasts that helps maintain 3D structure. The ability to reproducibly generate organoids that mimic the alveoli of the lung would have immense promise for studies aimed at understanding tissue function, the fundamental processes of respiratory infection, and the biomechanics of tissue structures during health and disease. Unfortunately, current protocols to generate alveolar organoids fail to reproduce native tissue structure. This RECODE project will uncover the rules necessary to differentiate alveolar progenitor cells into precise ratios of AT1:AT2 cells, and the contractile signaling that permits myofibroblasts to fold the epithelium into an alveolus. This transformational goal will be accomplished via a highly innovative combination of expertise from quantitative developmental biology, mechanobiology, biomaterials, computational modeling, and synthetic biology, which will be used to identify the spatiotemporal dynamics that governs alveolar differentiation and morphogenesis. The proposed research is divided into three main conceptual advances. Aim 1 focuses on using real-time fluorescent reporters, mathematical modeling, and optogenetics approaches to define the biochemical signaling dynamics necessary for specifying bipotent progenitors into AT2 cells. Aim 2 takes advantage of synthetic materials, real-time reporters, and optogenetics to uncover the mechanical signaling necessary for specifying bipotent progenitors into AT1 cells. Aim 3 combines computational modeling, 3D printing, and optogenetics to uncover and reproduce the patterns of contraction used by myofibroblasts to fold the epithelium into the alveolus. Altogether, this work will identify the design rules required to construct organoids that reproducibly differentiate into tissue structures that mimic alveoli within the lung. This RECODE award is co-funded by the Systems and Synthetic Biology Cluster in the Division of Molecular and Cellular Biosciences, the Developmental Systems Cluster in the Division of Integrative Organismal Systems, and the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental, and Transport Systems. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.