RECODE: Defining Environmental Design Criteria for Directed Differentiation of Type 1 from Type 2 Lung Alveolar Epithelial Cells

RECODE：定义 1 型肺泡上皮细胞与 2 型肺泡上皮细胞定向分化的环境设计标准

• 批准号: 2225554
• 负责人: Daniel Weiss
• 金额: $ 150万
• 依托单位: University of Vermont & State Agricultural College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2225554&HistoricalAwards=false
• 关键词: RECODE; Defining; Environmental; Design; Criteria

Injury to the lungs can have devastating consequences, as exemplified by the recent COVID19 pandemic. However, the cellular and molecular mechanisms by which the lung repairs itself remain poorly understood. The objective of this RECODE project is to utilize novel and sophisticated bioengineering approaches to better define cell and molecular pathways underlying lung development and repair. The project focuses on cells involved in the major function of the lung: gas exchange which provides oxygen to the body. This information will inform the development of new tunable biomaterials to guide lung cell development. The research efforts of this RECODE project are integrated with educational and outreach objectives to promote active learning in biomedical engineering and biologic sciences undergraduates, to develop outreach programs to encourage and inspire local high school science, engineering, and mathematical sciences students by hosting educational workshops poster sessions, and to promote biomedical engineering research and education towards the general public at each of the participating sites in Vermont, Colorado, and Iowa.There remains a critical need for better understanding of fundamental cellular and molecular mechanisms of lung development and repair, particularly with respect to the alveolar epithelium, a fundamental component of gas exchange. Current in vitro model systems, including organoid cultures, have provided important information but fail to fully reproduce native tissue structure or relevant environmental influences such as extracellular matrix (ECM) composition or stiffness. The central vision of this RECODE project is to devise and validate a robust system for delineating the mechanisms by which ECM composition and stiffness regulate differentiation of alveolar type 2 epithelial cells (AT2s) to alveolar type 1 epithelial cells (AT1s). Utilizing AT2s derived from human induced pluripotent stem cells (iAT2s), sophisticated tissue engineering approaches incorporating hydrogels derived from alveolar-enriched regions (aECM) of decellularized human lungs will be developed to evaluate effects of physiologically relevant ECM composition and stiffness on AT2 to AT1 directed differentiation. In silico modeling will be deployed in parallel to direct the empiric studies and to develop a holistic differentiation control framework. These approaches will be assessed in specific directed objectives: 1) To determine the specific ECM components regulating primary vs iAT2 stemness and driving AT1 differentiation; 2) To investigate the impact of dynamically tunable microenvironmental stiffness on primary vs iAT2 stemness and AT1 differentiation; and 3) To leverage agent-based and statistical modeling to predict combinatorial effects of composition and stiffness on primary vs iAT2 to AT1 differentiation. These unique and innovative approaches involve a multidisciplinary and multi-institutional combination of materials science, lung regenerative medicine, lung stem cell biology, and in silico modeling. Further, the paradigms and approaches generated will have broader impact and applicability to understanding cell-ECM interactions in enabling cell differentiation in a wider range of organ systems.This RECODE project is jointly funded by the Engineering Biology and Health Cluster in the Division of Chemical, Bioengineering, Environmental, and Transport Systems, the Established Program to Stimulate Competitive Research (EPSCoR), and the Physiological Mechanisms and Biomechanics Program and Animal Developmental Mechanisms Program in the Division of Integrative Organismal 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.

肺部损伤可能会造成毁灭性后果，最近的新冠肺炎大流行就是一个例子。 然而，人们对肺部自我修复的细胞和分子机制仍知之甚少。 该 RECODE 项目的目标是利用新颖且复杂的生物工程方法来更好地定义肺部发育和修复的细胞和分子途径。 该项目的重点是参与肺部主要功能的细胞：为身体提供氧气的气体交换。 这些信息将为新型可调节生物材料的开发提供信息，以指导肺细胞的发育。 RECODE 项目的研究工作与教育和外展目标相结合，以促进生物医学工程和生物科学本科生的主动学习，制定外展计划，通过举办教育研讨会海报来鼓励和启发当地高中科学、工程和数学科学学生会议，并在佛蒙特州、科罗拉多州和爱荷华州的每个参与地点促进对公众的生物医学工程研究和教育。仍然迫切需要更好地了解肺部发育和修复的基本细胞和分子机制，特别是尊重肺泡上皮，气体交换的基本组成部分。当前的体外模型系统，包括类器官培养物，已经提供了重要信息，但无法完全再现天然组织结构或相关环境影响，例如细胞外基质（ECM）组成或硬度。该 RECODE 项目的中心愿景是设计和验证一个强大的系统，用于描述 ECM 成分和硬度调节肺泡 2 型上皮细胞 (AT2s) 向肺泡 1 型上皮细胞 (AT1s) 分化的机制。 利用源自人诱导多能干细胞 (iAT2) 的 AT2，将开发复杂的组织工程方法，结合源自脱细胞人肺的肺泡富集区 (aECM) 的水凝胶，以评估生理相关 ECM 成分和硬度对 AT2 至 AT1 定向的影响差异化。将并行部署计算机建模来指导经验研究并开发整体差异化控制框架。这些方法将在具体的定向目标中进行评估： 1) 确定调节原代干性与 iAT2 干性并驱动 AT1 分化的特定 ECM 成分； 2) 研究动态可调微环境刚度对原代干性、iAT2 干性和 AT1 分化的影响； 3) 利用基于主体的统计模型来预测成分和刚度对初级分化与 iAT2 至 AT1 分化的组合影响。 这些独特和创新的方法涉及材料科学、肺再生医学、肺干细胞生物学和计算机建模的多学科和多机构组合。 此外，所产生的范式和方法将对理解细胞-ECM相互作用产生更广泛的影响和适用性，从而使细胞在更广泛的器官系统中分化。该RECODE项目由化学部工程生物学和健康集群联合资助，生物工程、环境和运输系统、刺激竞争研究既定计划（EPSCoR）、生理机制和生物力学计划以及动物发育机制计划综合有机系统。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。