An integrated experimental and theoretical approach to understanding corneal epithelial maintenance

了解角膜上皮维护的综合实验和理论方法

• 批准号: BB/J015172/1
• 负责人: John West
• 金额: $ 34.36万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ015172%2F1
• 关键词: integrated; experimental; theoretical; approach; understanding

Maintenance of the cornea, the transparent front surface of the eye that acts as our window on the world, is necessary for normal vision. There are many ways in which it can get permanently damaged, leading to opacity and potential loss of sight. In spite of intense study by many laboratories, major questions relating to the maintenance of the corneal surface remain. For example, although a large body of evidence exists which implies that stem cells have a major role, the actual location of these stem cells, and to what extent and under what circumstances they are active, remains highly controversial. Similarly, while it is known that cell migration occurs throughout life in the cornea, its importance and even its direction have been disputed. Because maintenance of the corneal surface is fundamental to our eyesight it is essential that we understand the basic biology about how the cornea functions normally and reacts, in the short and long term, to injury.This project will address the significant issues about stem cell localisation and activity, cell production, migration and loss, within the corneal surface epithelium. We will build upon a computer model that we have already developed that potentially explains some of the patterns of cell migration and loss that we have previously observed. We will refine that model using new biological data, and then use it to make testable predictions about how the cornea is maintained normally, how it is affected by ageing and genetic defects and how it responds to injury. We have an array of 'reporter' mice which, because they express fluorescent or other chemical markers in subsets of their corneal epithelial cells, can be used to trace patterns of cell production and migration in the cornea throughout life, starting during gestation and continuing into the ageing adult. Using these mice, we will be able to model genetic abnormalities and corneal injuries that test the predictions of the computer model. The mice will be used to show where the stem cells are localised in the cornea - knowledge that is essential to our understanding of how the epithelium is maintained. They will show how normal patterns of cell production and migration in the corneal surface are changed by injury and genetic defects, and also reveal whether these changes are long or short term. Changes in stem cell activity and migration patterns with ageing will also be determined using these mice - it is known for example that ageing human corneas heal less quickly than those from younger people. The refined model will have lasting benefits, as we will be able to use it to predict how different types of abnormalities occur in different corneal diseases. It will also contribute to reducing future animal experiments. Overall the project will generate key fundamental understanding of the processes that underlie corneal maintenance during normal life. We will be able to determine where the stem cells are, what they do and how replacement cells get to sites of injury. This knowledge will also help us understand why the corneal surface sometimes suffers degeneration and opacification that is difficult to treat. If new insights from our animal studies are applicable to humans this may also improve the management of corneal diseases or complications arising from operations such as laser eye surgery.

角膜是眼睛透明的前表面，是我们观察世界的窗口，对其进行维护对于正常视力至关重要。有多种方式可能导致其永久性损坏，从而导致混浊和潜在的失明。尽管许多实验室进行了大量研究，但与角膜表面维护相关的主要问题仍然存在。例如，尽管存在大量证据表明干细胞具有重要作用，但这些干细胞的实际位置以及它们活跃的程度和情况仍然存在很大争议。同样，虽然众所周知细胞迁移发生在角膜的整个生命周期，但其重要性甚至方向一直存在争议。由于角膜表面的维护对我们的视力至关重要，因此我们必须了解角膜如何正常运作以及在短期和长期内如何对损伤做出反应的基本生物学。该项目将解决角膜表面上皮内干细胞定位和活性、细胞产生、迁移和损失的重大问题。我们将建立在我们已经开发的计算机模型的基础上，该模型可能解释我们之前观察到的一些细胞迁移和损失的模式。我们将利用新的生物数据完善该模型，然后用它来做出可测试的预测，了解角膜如何保持正常状态、它如何受到衰老和遗传缺陷的影响以及它如何对损伤做出反应。我们有一系列“报告”小鼠，因为它们在角膜上皮细胞的子集中表达荧光或其他化学标记，因此可用于追踪整个生命过程中角膜中细胞产生和迁移的模式，从妊娠期间开始一直持续到老年。使用这些小鼠，我们将能够模拟遗传异常和角膜损伤，以测试计算机模型的预测。这些小鼠将被用来显示干细胞在角膜中的定位——这对于我们了解上皮细胞的维持方式至关重要。他们将展示角膜表面细胞生成和迁移的正常模式如何因损伤和遗传缺陷而改变，并揭示这些变化是长期还是短期的。干细胞活性和迁移模式随衰老的变化也将通过这些小鼠来确定——例如，众所周知，衰老的人类角膜愈合速度比年轻人的角膜慢。完善的模型将具有持久的好处，因为我们将能够使用它来预测不同角膜疾病中不同类型的异常是如何发生的。它还将有助于减少未来的动物实验。总体而言，该项目将对正常生活中角膜维护的过程产生关键的基本理解。我们将能够确定干细胞在哪里、它们的作用以及替代细胞如何到达损伤部位。这些知识还将帮助我们理解为什么角膜表面有时会出现难以治疗的变性和混浊。如果我们的动物研究的新见解适用于人类，这也可能改善角膜疾病或激光眼科手术等手术引起的并发症的治疗。

项目成果

Abnormal corneal epithelial maintenance in mice heterozygous for the micropinna microphthalmia mutation Mp.

• DOI: 10.1016/j.exer.2016.05.021
• 发表时间: 2016-08
• 期刊: Experimental eye research
• 影响因子: 3.4
• 作者: Douvaras P;Dorà NJ;Mort RL;Lodge EJ;Hill RE;West JD
• 通讯作者: West JD

Identifying stem cell locations in the mouse corneal and limbal epithelia by lineage tracing

通过谱系追踪鉴定小鼠角膜和角膜缘上皮细胞中的干细胞位置

• 发表时间: 2015
• 期刊: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
• 影响因子: 4.4
• 作者: Dora Natalie J.

Computer simulation of neutral drift among limbal epithelial stem cells of mosaic mice.

• DOI: 10.1016/j.scr.2018.05.003
• 发表时间: 2018-07
• 期刊: Stem cell research
• 影响因子: 1.2
• 作者: West JD;Mort RL;Hill RE;Morley SD;Collinson JM
• 通讯作者: Collinson JM

Lineage tracing in the adult mouse corneal epithelium supports the limbal epithelial stem cell hypothesis with intermittent periods of stem cell quiescence.

• DOI: 10.1016/j.scr.2015.10.016
• 发表时间: 2015-11
• 期刊: Stem cell research
• 影响因子: 1.2
• 作者: Dorà NJ;Hill RE;Collinson JM;West JD
• 通讯作者: West JD

Evaluating alternative stem cell hypotheses for adult corneal epithelial maintenance.

• DOI: 10.4252/wjsc.v7.i2.281
• 发表时间: 2015-03
• 期刊: World journal of stem cells
• 影响因子: 4.1
• 作者: J. West;Natalie J. Dorà;J. Collinson