Gene Discovery in Mouse Models for Secondary Cataracts

描述（由申请人提供）：上皮细胞向间充质细胞转化（EMT）是白内障手术后的主要问题之一。粘附于透镜囊的透镜上皮细胞可以生长并分化成间充质细胞，导致后囊膜混浊（PCO）。这个过程导致继发性白内障，必须再次治疗。鉴定促进透镜上皮细胞转分化的因子对于PCO的可能治疗是至关重要的。不幸的是，这一领域的实验受到缺乏实验模型的阻碍，这些模型可以应用强大的遗传技术。我们已经发现，当在小鼠中进行囊外手术时，囊上剩余的透镜上皮细胞迅速增殖，并用正常外观的透镜纤维填充囊袋。还可以看到“再生”透镜的正常形态，具有建立的弓形区域。再生透镜的形态取决于手术和透镜囊的完整性。然而，沿着向透镜纤维的正常分化，透镜上皮细胞也可以转分化为间充质细胞，特别是在透镜再生的早期阶段。因此，这种透镜再生小鼠模型可以成为研究PCO不可缺少的和长期寻求的动物模型。利用这样的动物模型系统，我们建议研究全球基因表达模式和基因发现的过程中。为了实现这一点，我们将研究透镜再生的小鼠与突变的基因重要的EMT和PCO。同时，这些小鼠经历（或不经历）透镜再生或EMT的能力将与使用全小鼠基因组微阵列的基因表达相关。关键基因的表达和作用将在使用囊袋的功能测定中验证。将建立的数据库进行相互比较，并与从发育中、成人和白内障晶状体建立的数据库进行比较。这项研究将最终确定参与EMT和PCO以及正常透镜纤维分化的已知和新基因的基因模式。本研究旨在为继发性白内障的治疗和后囊膜混浊的消除建立新的方法。此外，这项研究将解决透镜上皮细胞的双能性，以进行EMT，这导致继发性白内障，或透镜纤维分化，这是一个正常的修复过程。

DESCRIPTION (provided by applicant): Epithelial to Mesenchymal Cell Transition (EMT) is one of the major problems following cataract surgery. Lens epithelial cells adherent to the lens capsule can grow and differentiate into mesenchymal cells resulting in posterior capsule opacification (PCO). This process leads to secondary cataracts that have to be treated again. Identification of factors that promote transdifferentiation of the lens epithelial cells is of paramount importance for possible therapies of PCO. Unfortunately, experiments in this field are hindered by the lack of experimental models where the powerful genetic techniques can be applied. We have found that when extracapsular surgery is performed in the mouse, the remaining lens epithelial cells on the capsule proliferate rapidly and fill the bag with normal looking lens fibers. Normal morphology of the "regenerated" lens can also be seen, with established bow regions. The morphology of the regenerated lens depends on the surgery and the integrity of the lens capsule. However, along with the normal differentiation to lens fibers, lens epithelial cells can also transdifferentiate to mesenchymal cells, especially at the early stages of lens regeneration. This lens regeneration mouse model can become, therefore, an indispensable and long-sought animal model to study PCO. Taking advantage of such an animal model system we propose to study global gene expression patterns and gene discovery during the process. In order to achieve this we will examine lens regeneration in mice with mutations in genes important to EMT and PCO. At the same time the ability of these mice to undergo (or not) lens regeneration or EMT will be correlated with gene expression using whole mouse genome microarrays. Expression and roles of key genes will be verified in functional assays using capsular bags. The databases that will be established will be compared with each other and with databases established from developing, adult and cataractous lenses. This research will eventually identify gene patterns of known and novel genes that are involved in EMT and PCO as well as in normal lens fiber differentiation. This research aims to establish new ways for the treatment of secondary cataracts and the elimination of posterior capsule opacification. Also, this research will address the bipotency of lens epithelial cells to undergo EMT, which results in secondary cataracts, or lens fiber differentiation, which is a normal repair process.