3D Bioprinting Engineering Artificial Respiratory Tract Tissue

建议的项目适合优先领域：前沿医疗保健背景：需要开发和改进与人类相关的工具，以便研究人类生物学中尚未完全理解的机制。目前的体内模型几乎完全利用非灵长类动物，这些动物受到人类和动物系统之间频繁的差相关性的限制。理想的模型将允许在不同的实验条件下直接询问和比较基因相同的人体组织。在体外培养人体组织方面的进展，导致了体外人体器官培养的发展。迄今为止，这主要集中在产生用于移植的人体器官上。然而，这些技术的发展也为人类生物学的体外研究提供了革命性的潜力。虽然脱细胞支架有望用于移植，但对人类或动物组织产生支架的要求限制了该技术在科学发现等其他领域的大规模应用。3D生物打印是创建空间控制细胞模式的过程，其中生物组织的行为可以被复制。这理想地扩展到打印完整的，可行的器官体外模型，组织修复和器官移植。打印可行的器官目前还不可能，因为器官非常复杂。大多数打印的组织结构不能存活很长时间，打印后可打印的细胞负载生物链接和结构灌注仍然是开放的问题。尽管存在诸多挑战，但利用生物3D打印技术已经制造出了具有功能的微型器官，具有进一步发展的前景。研究计划：该项目的目的是开发和优化工程人工支架，适用于离体人体呼吸道器官培养。我们建议研究利用先进的3D生物打印技术，在试管中实现器官生长的方法，最终目标是显著减少或完全取代动物器官在研究中的使用。具体而言，研究计划将包括：(i)打印人造人上皮组织1.2影响总结（最多1500个字符）1.3数据共享（最多2000个字符）2第一步是开发可打印的水凝胶和细胞负载生物墨水，用于制造气道组织。在气液界面模型中，我们将打印和培养成纤维细胞间质和上皮组成的多细胞结构的人工气道，以获得完全分化的上皮。将通过组织学分析和生物标志物分析研究打印支架内的细胞自组装，并将其与来自供体组织的人肺支架进行比较。（ii） 3D打印微型化全器官支架项目第二阶段将打印整合气道的人工肺组织，与在人体全器官支架上的人体细胞自组装相比。基于组织学、转录组学和蛋白质生物标志物的分析将用于比较使用微型支架生成的人工组织与整个供体组织。本研究中生成的组织将使用先前描述的组织功能介质进行测试（例如，在肺组织模型中辛伐他汀抑制上皮细胞气道粘液产生，或在肠道模型中白细胞介素1 β in抑制乙酰胆碱诱导的肠道运动）。所研究的特定功能介质将根据(i)和（ii）的结果进行定义。

The proposed project fits in Priority Area: Leading Edge HealthcareBackground:There is a need to develop and improve human relevant tools enabling the study of mechanisms not yet fully understood in human biology. Current in vivo models almost exclusively utilise non primate animals which are limited by frequent poor correlation between human and animal systems. The ideal model would allow the direct interrogation and comparison of genetically equivalent human tissues under different experimental conditions. Advances in growing human tissues ex vivo, have led to the development of ex vivo human organ culture. To date this has focused on generating human organs for transplantation. However, the techniques developed also offer the potential to revolutionise in vitro studies of human biology. Whilst decellularised scaffolds hold promise for transplantation, the requirement for human or animal tissue to generate scaffolds limits the broad scale application of the technology to other areas such as science discovery.3D bio-printing is the process of creating spatially-controlled cell patterns, in which the behaviour of biological tissues can be reproduced. This ideally extends to printing complete, viable organs for in vitro model, tissue repair and organ transplant. Printing viable organs is not possible as of yet, as organs are very complex. Most printed tissue constructs are not viable for very long, printable cell-laden bioink and perfusion of the construct after printing are still open issues. Despite challenges, the functioning micro- organs have been produced by 3D bioprinting, promising for further development.Research Plan:The aim of this project is to develop and optimise engineered artificial scaffolds amenable to ex vivo human respiratory tract organ culture. We propose to investigate approaches to enable the growth of organs in test tubes, by making use of advances in 3D bioprinting technologies with an ultimate goal to significantly reduce/entirely replace the use of animal organs in research. Specifically the programme of research would consist of:(i) Printing artificial human epithelial tissue1.2 IMPACT SUMMARY (Up to 1500 characters)1.3 DATA SHARING (Up to 2000 characters)2The first step is to develop printable hydrogels and cell loaded bio-ink for fabricating airway tissue. Artificial airway with multicellular structure consisting of fibroblast stroma with epithelium will be printed and cultured for growing a fully differentiated epithelium in air-liquid interface models. Cellular self-assembly within printed scaffold will be studied and compared with human lung scaffolds from donor tissue through histological analysis and biomarker profiling.(ii) 3D printing miniaturised whole organ scaffoldsThe 2nd stage of the project will print artificial lung tissue with integration of airway, compared with human cell self-assembly on human whole organ scaffolds. Histological, transcriptomic and protein biomarker based profiling would be used to compare artificial tissue generated using miniaturised scaffolds to whole donor tissue.(iii) Functional studies of miniaturised artificial tissueThe tissue generated within this study would be tested using previously described mediators of tissue function (e.g. inhibition of airway mucus production of epithelial cells by simvastatin in a lung tissue model, or inhibition of acetylcholine induced intestinal motility by interleukin 1 beta in in a gut model). The specific functional mediators investigated would be defined based on the outcome of (i) and (ii).