Reconstructing native soluble cues in vascularized whole lung scaffolds

重建血管化全肺支架中的天然可溶性线索

• 批准号: 10449810
• 负责人: Yifan Yuan
• 金额: $ 12.43万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-10 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449810
• 关键词: Acute Respiratory Distress Syndrome; Adult; Alveolus; BMP5 gene; Biological Models; Biomimetics; Bioreactors; Blood Vessels; COVID-19; Capillary Leak Syndrome; Cell Maturation; Cells; Characteristics; Cues; Data; Data Set; Development; Disease; Disease model; Distal; Drug Screening; Endothelial Cells; Endothelium; Engineering; Environment; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; Functional disorder; Genomics; Homeostasis; Human; In Vitro; Inflammation; Inhalation; Injury; Interferon Type II; Lipopolysaccharides; Lung; Lung diseases; Microfluidics; Modeling; Organ; Organoids; Paracrine Communication; Phase; Phenotype; Physiological; Physiology; Property; Publishing; Sepsis; Signal Transduction; Structure; System; TNF gene; Thrombus; Trees; Vascular Permeabilities; Work; base; computerized tools; cytokine; drug candidate; drug mechanism; drug testing; hemodynamics; improved; novel; novel coronavirus; paracrine; primary pulmonary hypertension; pulmonary vascular disorder; scaffold; shear stress; single-cell RNA sequencing; solute; tissue culture; tool; vascular factor

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.

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（COVID-19）， 由于缺乏功能性离体模型，难以研究。传统的培养系统通常 它们在疾病和药物机制研究中代表人类病理生理学的能力有限。的 该提案的总体目标是开发一种模拟肺动脉高压的实验平台。 微血管，并且可以在体外模拟天然细胞表型和功能，在正常 体内平衡和疾病状态如严重炎症和败血症期间。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成分都是 对于调节内皮成熟和维持血管内环境稳定至关重要。整个器官 去细胞化打开了一扇门， 一整棵血管树此外，利用单细胞RNA-seq（scRNAseq），我们开发了 计算工具来识别人类远端肺中的旁分泌信号。在这项研究中，我将利用这些 在功能性肺微血管生态位中鉴定新的、重要的局部作用可溶性因子的工具， 在无细胞肺支架中改善肺微血管成熟。在本提案的K99阶段， 我将首先利用我们已发表的scRNAseq计算工具来评估原生的scRNAseq数据集。 成年人的肺我将确定一组重要的和新颖的可溶性因子， 内皮成熟然后，我将理性地在我们的内皮再生肺平台上， 添加来源于天然微血管环境的相关可溶性因子。在R 00阶段，我将 利用前期建立的血管平台，开始开发疾病建模系统，研究 炎症对药物测试的影响。这项工作将导致创建一个新的平台，不像 先前的微血管平台特别类似于天然肺的许多生理方面 环境这种平台可用于肺血管疾病建模和药物测试。