CAREER: Decoding the Developmental Extracellular Niche for Instructing Lung Tissue Engineering

我们如何设计人体组织的灵感通常来自于这些组织在胚胎发育期间如何在体内被诱导，以及它们在受伤后如何再生。本CAREER项目的目的是揭示在肺组织发育和手术切除诱导再生过程中支持肺组织成熟的细胞外生物分子环境的动态变化。这些信息将用于指导新型细胞外生物材料的工程设计，以实现人类诱导多能干细胞（hiPSCs）在实验室环境下的定向肺组织成熟。通过识别和验证必需的细胞外因子，该项目将为获得功能性人肺组织奠定基础，以帮助终末期肺部疾病患者，并将激发促进损伤后肺修复和再生的新疗法。CAREER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课堂上的主动学习和自动学生行为分析，通过举办教育海报会议，制定外展计划，鼓励和激励当地高中生，并通过与当地一家专注于生物艺术的博物馆合作，向公众推广生物医学工程研究和教育。研究者的长期研究目标是设计出满足独特生理和功能需求的肺组织。为了实现这一目标，本CAREER项目的目标是对原生肺泡形成过程中细胞外基质（ECM）糖蛋白的动态合成和分泌进行分类，并利用这些信息指导hiPSCs诱导肺泡细胞和组织。新出现的证据表明，发育调节的ECM沉积对肺泡细胞的成熟和适当的肺组织形态发生至关重要。然而，由于检测新ECM蛋白产生的灵敏度有限，在整个肺发育和再生过程中对新ECM合成的全面调查一直具有挑战性。为了解决这一瓶颈，本研究将开发并实施一种新的新合成ECM （newsECM）分析方法，以揭示伴随肺泡形成的动态ECM合成程序。这种策略，通过选择性标记和富集新的ECM蛋白，使新的ECM合成具有前所未有的每日时间分辨率的超灵敏鉴定。这解决了当前ECM蛋白质组学的关键限制，非常适合研究高动态过程，如肺泡形成。在这项技术创新的基础上，这个CAREER项目将(1)解码原生肺泡形成（新生儿和成人肺切除术后）过程中动态ECM的产生，(2)利用发育启发的ECM支架设计hipsc衍生的肺泡形成。从hipsc中获得定向肺泡规格将为肺发育提供基本见解，并促进患者特异性肺组织的产生，用于药物发现和再生医学。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。

How we engineer human tissues is usually inspired by how these tissues are induced in the body during embryonic development and how they regenerate following an injury. The objective of this CAREER project is to uncover the dynamic changes in the extracellular biomolecular environment that support lung tissue maturation during development and during surgical resection induced regeneration. This information will then be used to guide the engineering of novel extracellular biomaterials to enable directed lung tissue maturation in the laboratory setting from human induced pluripotent stem cells (hiPSCs). By identifying and validating the essential extracellular factors, this project will lay the foundation for deriving functional human lung tissues to help patients suffering from end-stage lung diseases and will inspire novel therapeutics promoting lung repair and regeneration following injuries. The research efforts of the CAREER project are integrated with educational and outreach objectives to promote active learning and automated student behavior analysis in the biomedical engineering classroom, to develop an outreach program to encourage and inspire local high school students by hosting educational poster sessions, and to promote biomedical engineering research and education towards the general public through collaboration with a local museum focusing on biological arts.The investigator's long-term research goal is to engineer lung tissues that meet unique physiological and functional requirements. Towards this goal, the objective of this CAREER project is to catalog the dynamic synthesis and secretion of extracellular matrix (ECM) glycoproteins over native lung alveologenesis and use this information to instruct the induction of alveolar cells and tissues from hiPSCs. Emerging evidence suggests that the developmentally regulated ECM deposition is pivotal to the maturation of alveolar cells and proper lung tissue morphogenesis. However, a comprehensive survey of de novo ECM synthesis throughout lung development and regeneration has been challenging to obtain, due to the limited sensitivity in detecting the production of new ECM proteins. To address this bottleneck, this research will develop and implement a novel newly synthesized ECM (newsECM) profiling approach for uncovering the dynamic ECM synthetic programs accompanying alveologenesis. This strategy, by selective labeling and enrichment of newsECM proteins free from the bulk pre-existing ECM, will enable ultrasensitive identification of new ECM synthesis with unprecedented daily temporal resolution. This addresses the critical limitation of current ECM proteomics and is well suited for studying highly dynamic processes such as alveologenesis. Building on this technological innovation, this CAREER project will (1) decode the dynamic ECM production during native lung alveologenesis (neonatal and adult post-pneumonectomy), and (2) engineer hiPSC-derived alveologenesis using developmentally inspired ECM scaffolds. Achieving directed alveolar specification from hiPSCs will provide fundamental insights regarding lung development and facilitate the generation of patient-specific lung tissues for drug discovery and regenerative medicine.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.