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

RECODE：利用光和力学监测和控制肺泡类器官的分化

• 批准号: 2134935
• 负责人: Celeste Nelson
• 金额: $ 150万
• 依托单位: Princeton University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2134935&HistoricalAwards=false
• 关键词: RECODE; Using; light; mechanics; monitor

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.

细胞可以在体外被诱导形成类似器官的结构。这些“类器官”将帮助科学家研究器官的发育、功能和疾病。类器官通常不能以可复制的方式形成，目前限制了它们的效用。这个项目将发明新的方法来可靠地构建模拟肺的类器官。研究人员将利用光和机械力跟踪和控制类器官的形成。该项目将把来自弱势群体的高中生和大学生引入科学研究。该项目还将通过建立一个国际研讨会来分享其方法。肺的最小功能单位是肺泡，它由肺泡上皮I型（AT1）细胞和II型（AT2）细胞组成，由肌成纤维细胞网包围，有助于维持三维结构。可复制地生成模仿肺泡的类器官的能力将为旨在理解组织功能、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带来巨大的希望。不幸的是，目前生成肺泡类器官的方法无法复制原生组织结构。RECODE项目将揭示肺泡祖细胞分化为AT1:AT2细胞的精确比例所必需的规则，以及允许肌成纤维细胞将上皮折叠成肺泡的收缩信号。这一转变目标将通过定量发育生物学、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确定控制肺泡分化和形态发生的时空动态。提出的研究分为三个主要的概念进展。目的1侧重于使用实时荧光报告，数学建模和光遗传学方法来定义将双能祖细胞指定为AT2细胞所需的生化信号动力学。Aim 2利用合成材料、实时报告和光遗传学来揭示将双能祖细胞指定为AT1细胞所必需的机械信号。目的3结合计算建模、3D打印和光遗传学来揭示和重现肌成纤维细胞将上皮折叠成肺泡所用的收缩模式。总之，这项工作将确定构建可重复分化为模仿肺内肺泡的组织结构的类器官所需的设计规则。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、综合有机体系统部门的发育系统集群以及化学、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。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Control of gastruloid patterning and morphogenesis by the Erk and Akt signaling pathways

Erk 和 Akt 信号通路对原肠胚模式和形态发生的控制

• DOI: 10.1242/dev.201663
• 发表时间: 2023
• 期刊: Development
• 影响因子: 4.6
• 作者: Underhill, Evan J.;Toettcher, Jared E.
• 通讯作者: Toettcher, Jared E.