3D Bioprinting Engineering Artificial Respiratory Tract Tissue

拟议的项目符合优先领域：前沿医疗背景：有必要开发和改进人类相关的工具，使人类生物学尚未完全理解的机制的研究。目前的体内模型几乎完全利用非灵长类动物，这受到人类和动物系统之间频繁的不良相关性的限制。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等同的人体组织。离体培养人体组织的进展导致了离体人体器官培养的发展。到目前为止，这一直集中在产生用于移植的人体器官。然而，开发的技术也提供了彻底改变人类生物学体外研究的潜力。虽然脱细胞支架有望用于移植，但人类或动物组织生成支架的要求限制了该技术在其他领域的广泛应用，例如科学发现。3D生物打印是创建空间控制的细胞模式的过程，其中生物组织的行为可以复制。这理想地扩展到打印完整的、可存活的器官，用于体外模型、组织修复和器官移植。打印可存活的器官目前还不可能，因为器官非常复杂。大多数打印的组织构建体不能存活很长时间，可打印的载有细胞的生物墨水和打印后构建体的灌注仍然是悬而未决的问题。尽管面临挑战，但功能性微器官已经通过3D生物打印产生，有希望进一步发展。研究计划：本项目的目的是开发和优化适合体外培养人类呼吸道器官的工程人工支架。我们建议通过利用3D生物打印技术的进步，研究在试管中生长器官的方法，最终目标是显着减少/完全取代研究中使用的动物器官。具体而言，研究计划将包括：（i）打印人造人类上皮组织1.2影响概要（最多1500个字符）1.3数据共享（最多2000个字符）2第一步是开发用于制造气道组织的可打印水凝胶和载有细胞的生物墨水。将打印和培养具有由具有上皮的成纤维细胞基质组成的多细胞结构的人工气道，以在气液界面模型中生长完全分化的上皮。将研究打印支架内的细胞自组装，并通过组织学分析和生物标志物分析与来自供体组织的人肺支架进行比较。(ii)该项目的第二阶段将打印具有气道整合的人工肺组织，与人体细胞在人体整个器官支架上的自组装相比。将使用基于组织学、转录组学和蛋白质生物标志物的分析来比较使用固定化支架产生的人工组织与整个供体组织。(iii)人造组织的功能研究本研究中产生的组织将使用先前描述的组织功能介质（例如，在肺组织模型中辛伐他汀对上皮细胞的气道粘液产生的抑制，或在肠模型中白细胞介素1 β对乙酰胆碱诱导的肠运动的抑制）进行测试。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).