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

我们如何设计人类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。这个职业项目的目标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。这些信息将被用于指导新型细胞外生物材料的工程设计，使人诱导多能干细胞(HiPSCs)能够在实验室环境下定向成熟肺组织。通过鉴定和验证必要的细胞外因子，该项目将为获得功能性人类肺组织以帮助终末期肺部疾病患者奠定基础，并将启发促进损伤后肺修复和再生的新疗法。该职业项目的研究努力与教育和推广目标相结合，以促进生物医学工程课堂上的主动学习和自动化学生行为分析，开发一个推广计划，通过举办教育海报会议来鼓励和启发当地高中生，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。研究人员的长期研究目标是设计满足独特生理和功能要求的肺组织。为了达到这一目标，本职业项目的目标是对天然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从HiPSCs诱导肺泡细胞和组织。新的证据表明，发育调节的细胞外基质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，由于检测新的ECM蛋白产生的灵敏度有限，对整个肺发育和再生过程中从头合成ECM的全面调查一直是具有挑战性的。为了解决这一瓶颈，本研究将开发和实现一种新的合成ECM(新闻ECM)剖析方法，以揭示伴随着肺泡形成的动态ECM合成程序。这一策略通过选择性地标记和浓缩不含大量预先存在的ECM的新闻ECM蛋白，将能够以前所未有的每日时间分辨率超灵敏地识别新的ECM合成。这解决了目前ECM蛋白质组学的关键局限性，非常适合于研究高度动态的过程，如肺泡形成。在这项技术创新的基础上，这个职业项目将(1)解码自然肺泡形成(新生儿和成人全肺切除后)过程中动态ECM的产生，以及(2)利用发育灵感的ECM支架设计HiPSC衍生的肺泡形成。从HiPSC获得定向的肺泡规范将提供关于肺发育的基本见解，并促进为药物发现和再生药物生成患者特定的肺组织。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。

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.