Deciphering molecular pathways and cellular events involved in uveal coloboma

什么是Coloboma？Coloboma指的是一种影响大约1：10,000儿童的眼部疾病。当眼睛在出生前发育时，它通常会闭合，形成一个球体。在缺陷症的情况下，球形眼睛没有完全形成，在其结构上留下了较小或更大的缝隙，通常是在眼睛的底部。这可能会导致儿童失明或部分视力。缺陷症的病因尚不完全清楚，但已知环境和遗传因素都是造成这种情况的原因。然而，大多数缺陷症似乎是由胚胎发育过程中的错误引起的，这种错误被称为突变，我们的遗传成分被称为基因。然而，仍然有许多缺陷症的致病基因尚未确定，正在投入巨大的努力来确定与这种情况有关的更多基因。由于缺陷起源于胎儿时期，研究胚胎发育过程中细胞之间的相互作用以及哪些基因参与了闭眼，有助于我们了解这种缺陷的原因。研究目的本研究的主要目的是找出可能导致人类缺陷的新基因和细胞事件。老鼠是研究人类遗传疾病的一个很好的模型，因为它与人类有许多共同的基因。在许多情况下，这些共享的基因包含人类和小鼠的错误(突变)，导致类似的缺陷。人类和小鼠的一些与缺陷相关的基因也是如此。目标-我们的第一个目标是从小鼠胚胎的眼睛底部分离出不同的基因集。这些基因对闭眼有潜在的关键作用。我们的策略将比较正常胚胎和FOXG1基因突变胚胎样本中表达的基因，这些胚胎有严重的缺陷。这种方法将产生大量的缺陷性候选基因，我们将需要优先处理这些基因。接下来将遵循几个步骤来缩小感兴趣基因的范围。在爱丁堡大学生物信息学家的帮助下，我们的结果将根据两组研究之间的显着差异进行排名。我们将把我们的结果与筛查缺陷症患者的结果进行比较。这将与爱丁堡MRC-人类遗传学研究所的缺陷症专家菲茨帕特里克教授合作实现。3.我们将调查在发育中的小鼠眼睛的哪些区域发现了我们最好的候选基因。然后，我们将研究阻断基因功能或增加这些候选基因的剂量对小鼠发育眼培养的影响。将从这项研究中产生的可能的缺陷性候选基因将在人类缺陷症患者的样本上进行进一步的突变测试。这将再次与Fitzpatrick教授的团队合作实现。-我们最近观察到，在FOXG1突变小鼠中，当我们减去另一种名为Wnt8b的基因时，我们能够改善缺陷性缺陷。这表明Wnt8b参与了缺陷的形成，在哺乳动物中减少该基因的数量可能会修复缺陷。我们的第二个目标是了解Wnt8b基因在眼睛发育过程中的作用，它与FOXG1以及其他导致小鼠和人类皮肤缺损的基因的关系，如Pax2基因。这项研究的好处我们期望了解修复修复背后的基本生物学事件，并识别与人类修复修复有关的新基因。这一发现将对更广泛的科学界具有重要意义。我们的基础研究将产生数据，这些数据将在未来帮助眼科医生、儿科医生和遗传学家为有缺陷症儿童的家庭提供更好的信息和支持。

What is Coloboma?Coloboma refers to an eye disorder that affects approximately 1:10,000 children. As the eye develops before birth it will normally close forming a sphere. In the case of coloboma the spherical eye does not fully form leaving small or bigger gaps in its structure, usually in the bottom part of the eye. These can result in the child being blind or partially sighted.Aetiology of Coloboma The causes of coloboma are not fully understood, but it is known that both environmental and genetic factors contribute to it. However, most cases of coloboma seem to be caused during embryonic development by errors, known as mutations, in our hereditary components known as genes. However, there are still many cases of coloboma for which the causative gene has not yet been identified and great effort is being invested in identifying additional genes involved in this condition. As coloboma originates during foetal life, investigating how the cells interact and what genes participate in the closure of the eye during embryonic development can help us understand the causes of this disorder.Research AimThe main aim of this research proposal is to identify new genes and cellular events that may be responsible for coloboma in humans.To achieve this aim we will study eye development using the mouse as our model organism. The mouse is a good model for studying human genetic disorders as it shares many common genes with humans. In many cases, these shared genes contain errors (mutations) in both human and mouse which result in similar defects. This is also the case for some of the genes associated with coloboma in both human and mouse.Objectives- Our first objective is to isolate different sets of genes from the bottom part of the eye of mouse embryos. These genes are potentially crucial for eye closure. Our strategy will compare genes expressed in samples of normal embryos and embryos that are mutant for the Foxg1 gene, which have severe coloboma. This approach will produce a lot of coloboma-candidate genes which we will need to prioritise. Several steps will be followed for narrowing down genes of interest.1. With help from Bioinformaticians from the University of Edinburgh our results will be ranked based on significant differences between our two groups of study.2. We will compare our results with those generated from a screening of patients with coloboma. This will be achieved in collaboration with Prof FitzPatrick, an expert in coloboma at the MRC-Human Genetics Unit in Edinburgh. 3. We will investigate in which areas of the developing mouse eye our best candidate genes are found.4. We will then investigate what are the effects on the mouse of blocking the gene function or increasing the dose of these candidate genes on cultures of mouse developing eyes.5. The genes that will arise from this study as plausible candidates for coloboma will be further tested for mutations on human samples of patients with coloboma. This, again, will be achieved in collaboration with Prof FitzPatrick's group.- We have recently observed that in the Foxg1 mutant mouse we are able to ameliorate the coloboma defect when we subtract another gene known as Wnt8b. This indicates that Wnt8b is involved in coloboma formation and reducing the amount of this gene in mammals may recover the defect. Our second objective is to understand how the Wnt8b gene acts during eye development, its relationship with Foxg1 and with other genes that cause coloboma in mice and human, such as the Pax2 gene. Benefits from the studyWe anticipate to understand the basic biological events behind coloboma and to identify novel genes responsible for human coloboma. Such a discovery will be of great importance to the wider scientific community. Our basic research will generate data that in the future will help ophthalmologists, paediatricians and geneticists to provide better information and support to families with children with coloboma.