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

细胞可以被诱使在体外形成类似器官的结构。这些“有机物质”将帮助科学家研究器官发育、功能和疾病。有机化合物通常不能以可重复的方式形成，目前限制了它们的用途。这个项目将发明新的方法来可靠地制造模仿肺的有机化合物。调查人员将使用光和机械力跟踪和控制有机物的形成。该项目将向来自代表性不足社区的高中生和大学生介绍科学研究。该项目还将通过建立国际研讨会来分享其方法。肺泡是肺的最小功能单位，由肺泡I型上皮(AT1)细胞和II型(AT2)细胞组成，周围环绕着帮助维持3D结构的肌成纤维细胞网络。可再生地产生模仿肺泡的有机物质的能力将在旨在了解组织功能、呼吸道感染的基本过程以及健康和疾病期间组织结构的生物力学的研究中具有巨大的前景。不幸的是，目前产生肺泡有机体的方案不能重现天然的组织结构。这个重新编码项目将揭示将肺泡祖细胞分化为精确比例的AT1：AT2细胞所必需的规则，以及允许肌成纤维细胞将上皮折叠成肺泡的收缩信号。这一变革性的目标将通过数量发育生物学、机械生物学、生物材料、计算建模和合成生物学等专业知识的高度创新组合来实现，这些专业知识将用于识别控制肺泡分化和形态发生的时空动力学。拟议的研究分为三个主要的概念性进展。目的1着重于使用实时荧光报告、数学建模和光遗传学的方法来定义在AT2细胞中指定双能祖细胞所必需的生化信号动力学。目标2利用合成材料、实时报告和光遗传学的优势，揭示了将双能祖细胞指定为AT1细胞所必需的机械信号。AIM 3结合了计算建模、3D打印和光遗传学，以揭示和复制肌成纤维细胞用来将上皮折叠到肺泡中的收缩模式。总之，这项工作将确定构建有机化合物所需的设计规则，这些有机化合物可在肺内重复分化为模仿肺泡的组织结构。该Recode奖由分子和细胞生物科学部的系统和合成生物学集群、集成组织系统部门的开发系统集群和化学、生物工程、环境和运输系统部门的工程生物学和健康集群共同资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.