Trp63 in limbal stem cell deficiency

项目摘要 转录因子tp63(人)/trp63(小鼠)，在本申请中被称为p63，是一个关键 复层上皮细胞发育的调节剂。完全缺乏p63表达的小鼠无法 形成复层上皮，包括角膜上皮。然而，p63也需要维持 并在出生后修复复层上皮。这一点在患者的临床表型中得到了强调。 患有指外翻、外胚层发育不良和唇腭裂综合征[欧共体；OMIM#604292]，a 由p63基因突变引起的疾病。EEC患者发生角膜缘干细胞缺乏症(LSCD)， 以进行性角膜侵蚀、角膜结膜形成和终末期为特征的一种状态 失明。除了p63异常和LSCD之间有明确的因果联系外，基本上没有什么是 已知依赖突变体p63的分子和细胞机制 病理学。此外，还不知道眼睛中的哪个细胞间隔(S)受到EEC的影响。我们 假设维持角膜缘干细胞(LSC)和驱动正常的P63功能是必需的 将这些细胞分化为角膜上皮细胞。为了检验这一假设，我们建议使用 基因工程小鼠模型和体外细胞培养模型相结合鉴定突变 眼上皮细胞中p63调控的基因调控通路。这一方法不仅将推动 角膜上皮干细胞生物学的基本知识，它还将指出可能 靶向治疗P63功能受损所致的LSCD患者。

Project Summary The transcription factor TP63 (human)/Trp63 (mouse), referred to as “p63” in this application, is a key regulator for the development of stratified epithelia. Mice that completely lack p63 expression fail to develop stratified epithelia, including the corneal epithelium. However, p63 is also required to maintain and repair stratified epithelia postnatally. This is underscored by the clinical phenotype of patients suffering from ectrodactyly, ectodermal dysplasia, and cleft lip/palate syndrome [EEC; OMIM #604292], a disorder caused by mutations in the p63 gene. EEC patients develop limbal stem cell deficiency (LSCD), a condition that is characterized by progressive corneal erosions, corneal conjunctivalization, and ultimate loss of vision. Besides a clear causal link between p63 abnormalities and LSCD, essentially nothing is known regarding the molecular and cellular mechanisms that underlie the mutant p63-dependent pathology. Further, it is not known which cell compartment(s) in the eye are affected by EEC. We hypothesize that normal p63 function is required to maintain limbal stem cells (LSC) and to drive proper differentiation of these cells into corneal epithelial cells. To test this hypothesis, we propose to use a combination of genetically engineered mouse models and in vitro cell culture models to identify mutant p63-controlled gene regulatory pathways in the ocular epithelium. This approach will not only advance basic knowledge of cornea epithelial stem cell biology, it will also point to molecular pathways that could be targeted therapeutically in patients suffering from LSCD caused by impaired p63 function.