Reconstructing native soluble cues in vascularized whole lung scaffolds

项目摘要 肺血管疾病，包括毛细血管渗漏综合征/急性呼吸窘迫综合征 (ARDS)、吸入性损伤、原发性肺动脉高压和新型冠状病毒病-19(新冠肺炎)， 由于缺乏有功能的体外模型，很难进行研究。传统的文化体系通常是 它们代表人类病理生理学研究疾病和药物机制的能力有限。这个 这一提议的总体目标是开发一个模拟肺组织的实验平台 微血管系统，可以在体外模拟正常情况下的自然细胞表型和功能 动态平衡和疾病状态期间，如严重炎症和败血症。肺微血管 生态位特征，如旁分泌因子、血流动力学和细胞外基质组成都是 对调节血管内皮细胞成熟和维持血管内环境稳定至关重要。整个器官 脱细胞打开了一扇门，以提供一种重塑衬底结构和成分的结构 一棵完整的维管树。此外，利用单细胞RNA-seq(ScRNAseq)，我们开发了 识别人类远端肺旁分泌信号的计算工具。在本研究中，我将利用这些 在功能性肺微血管生态位中识别新的、重要的局部作用的可溶性因子的工具 促进无细胞肺支架内肺微血管成熟。在本提案的K99阶段， 我将首先利用我们发布的scRNAseq计算工具在本机上评估scRNAseq数据集 成人的肺。我将确定一组重要的和新的可溶性因子，它们可以改善 血管内皮细胞成熟。然后，我将理性地迭代我们的内皮重新填充的肺平台 添加来自天然微血管环境的相关可溶性因子。在R00阶段，我将 利用已有的AIMS建立的血管平台，着手开发疾病建模系统，以研究 炎症对药检的影响。这项工作将导致创建一个新的平台，不同于 以前的微血管平台，特别类似于天然肺的许多生理方面 环境。该平台可用于肺血管疾病的建模和药物测试。

Project Abstract Diseases of the lung vasculature, including capillary leak syndrome/acute respiratory distress syndrome (ARDS), inhalation injury, primary pulmonary hypertension, and the novel coronavirus disease-19 (COVID-19), are difficult to study due to the lack of functional ex vivo models. Conventional culture systems are typically limited in their ability to represent human pathophysiology for the study of disease and drug mechanisms. The overall objective of this proposal is to develop an experimental platform that mimics the pulmonary microvasculature, and that can simulate native cellular phenotypes and functions in vitro, during normal homeostasis and during disease states such as severe inflammation, and sepsis. The lung microvascular niche characteristics, such as paracrine factors, hemodynamics, and extracellular matrix composition are all of pivotal importance for regulating endothelial maturation and maintaining vascular homeostasis. Whole organ decellularization opens a door to provide a construct that recapitulates the substrate structure and components of an entire vascular tree. Additionally, leveraging single-cell RNA-seq (scRNAseq), we developed computational tools to identify the paracrine signals in human distal lungs. In this study, I will leverage these tools to identify novel, important locally acting soluble factors in a functional lung microvascular niche to improve pulmonary microvascular maturation in acellular lung scaffolds. During the K99 phase of this proposal, I will first leverage our published scRNAseq computational tools to evaluate the scRNAseq dataset on native adult human lungs. I will determine a group of important and novel soluble factors that could improve endothelial maturation. Then, I will rationally iterate on our endothelial repopulated lung platform with the addition of relevant soluble factors derived from the native microvascular milieu. During the R00 phase, I will use the vascular platform established in prior aims and start to develop a disease modeling system to study the impact of inflammation for drug testing. This work will lead to the creation of a novel platform which, unlike previous microvascular platforms, specifically resembles many physiological aspects of the native lung environment. Such a platform could be used for pulmonary vascular disease modeling and drug testing.