Elucidating the role of disparities in the oxygen microenvironment in organoids and engineered tissues

阐明类器官和工程组织中氧微环境差异的作用

• 批准号: RGPIN-2022-03553
• 负责人: Ungrin, Mark
• 金额: $ 3.5万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=766033
• 关键词: Elucidating; role; disparities; oxygen; microenvironment

Local oxygen concentration is a primary driver of cell behaviour in vitro and in vivo, but is surprisingly difficult to assess and interpret even in simple cell culture systems. The role of oxygen in vivo has been extensively studied under normal conditions, and in disease states including myocardial and cerebral ischemia, wound healing and infection, although this is primarily at the tissue level. Considerably less is known about the hyper-local distribution of oxygen at the scale of the individual cell, how it affects their behaviour, and how oxygen delivery and consumption is impacted by microscopic structures such as epithelia, and extracellular matrices. In normal living tissues, blood vessels constantly constrict, relax, grow and are remodelled, while breathing and heart rates adjust continuously, and red blood cells are created and broken down - providing cells with a consistent and finely-tuned supply of oxygen. In contrast, cells in culture of the same type, in the same incubator, can experience oxygen levels ranging from far above to far below normal, due only to differences in density and medium volume. Just as water thrown on a fire provides a barrier between fuel and air, the medium in which most cell and tissue cultures are submerged significantly impacts their access to oxygen. Cell density, metabolic rate, and medium depth are critical variables - and these can all change over the course of an experiment. Cutting-edge research increasingly requires more accurate models of biological systems. 3D cultured tissues known as "organoids" are a powerful and popular tool for this purpose, however their complexity adds to both the challenge and importance of understanding the role of oxygen in their behaviour. The funding requested here will allow us to establish genetically-encoded oxygen reporter constructs, introduce them into diverse organoid types, calibrate them, and apply them to understand the role of oxygen in those organoids. We will generate very low-cost biological monitoring systems, and educate trainees in their use, and in the philosophy of research reproducibility that underlies them. Internationally, researchers in a wide range of disciplines have demonstrated the effectiveness of this strategy by adopting my approaches and "AggreWell" organoid manufacturing platform as standard methods. Our progress towards filling this knowledge gap will be broadly useful to this large and rapidly growing audience. This program is part of my long-term vision of making organoid culture systems widely accessible, consistent and reproducible, to accelerate research in the biological sciences around the world. In addition to publishing our findings, the novel tools we develop will be distributed globally, and benefit cell and tissue culture research (and the biomanufacturing industry being built upon it) in Canada and around the world, following in the footsteps of my previous successes at this level.

局部氧浓度是体外和体内细胞行为的主要驱动因素，但即使在简单的细胞培养系统中也难以评估和解释。氧在体内的作用已在正常条件下以及在包括心肌和脑缺血、伤口愈合和感染的疾病状态下被广泛研究，尽管这主要是在组织水平。关于氧在单个细胞尺度上的超局部分布，它如何影响它们的行为，以及氧的输送和消耗如何受到上皮细胞和细胞外基质等微观结构的影响，人们知之甚少。在正常的活组织中，血管不断收缩，放松，生长和重塑，同时呼吸和心率不断调整，红细胞被创造和分解-为细胞提供一致和微调的氧气供应。相比之下，在同一培养箱中培养的相同类型的细胞可以经历从远高于正常到远低于正常的氧水平，这仅仅是由于密度和培养基体积的差异。就像水在火上提供了燃料和空气之间的屏障一样，大多数细胞和组织培养物浸没在其中的介质会显著影响它们获得氧气的能力。细胞密度、代谢率和培养基深度是关键变量--这些都可能在实验过程中发生变化。尖端研究越来越需要更精确的生物系统模型。被称为“类器官”的3D培养组织是用于此目的的强大而流行的工具，然而它们的复杂性增加了理解氧气在其行为中的作用的挑战和重要性。这里要求的资金将使我们能够建立遗传编码的氧报告基因构建体，将它们引入不同的类器官类型，校准它们，并应用它们来了解氧在这些类器官中的作用。我们将生产成本非常低的生物监测系统，并教育学员如何使用它们，以及作为其基础的研究可重复性的哲学。在国际上，广泛学科的研究人员已经通过采用我的方法和“AggreWell”类器官制造平台作为标准方法来证明了这一策略的有效性。我们在填补这一知识空白方面取得的进展将对这一庞大且迅速增长的受众广泛有用。该计划是我长期愿景的一部分，即使类器官培养系统广泛可用，一致和可重复，以加速世界各地生物科学的研究。除了发表我们的研究结果，我们开发的新工具将在全球范围内分发，并有利于加拿大和世界各地的细胞和组织培养研究（以及建立在此基础上的生物制造业），追随我以前在这一水平上取得成功的脚步。