Limbal Stem Cell Fate and Corneal Specific Enhancers

角膜缘干细胞命运和角膜特异性增强剂

基本信息

批准号：
9243258
负责人：
GEORGE L SEN
金额：
$ 47.41万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9243258
关键词：
Address Affect Alpha Cell Animal Model Biological Biological Assay Biology Blindness Cell Differentiation process Cells Cellular biology ChIP-seq Characteristics Chromatin Cornea Cultured Cells DNA Development Disease Down-Regulation Enhancers Epigenetic Process Epithelial Epithelial Cells Epithelium Etiology Eye Eye Development Gene Expression Gene Expression Regulation Generations Genes Genetic Genetic Transcription Genetic Variation Genomics Goals High-Throughput Nucleotide Sequencing Histones Human Infection Inflammation Keratin Knowledge Mediating Medical Molecular Mutation Play Predisposition Reagent Regulation Regulatory Element Reporter Repression Resources Role Skin Specificity Stem cells Surface Testing Tissue-Specific Gene Expression Tissues To specify Transplantation Trauma Vision Visual impairment Work base cell type corneal epithelium epigenetic regulation genome-wide analysis human disease interdisciplinary approach keratinization limbal progenitor public health relevance stem cell biology stem cell fate tool transcription factor translational study

项目摘要

DESCRIPTION (provided by applicant): The cornea is a specialized epithelium, which protects the eye and maintains transparency for normal vision. Diseases of the cornea or its stem cells (limbal stem cells) affects millions of people worldwide and result from a spectrum of etiologies, ranging from genetic defects to infection, inflammation, or trauma. Vision loss often occurs as the result of transformation of the transparent cornea into a skin-like epithelium. Recent work from our labs and others now reveal that this critical transition may be precipitated by the loss of PAX6 expression. Downregulation of PAX6 causes the loss of cornea features and gene expression and the ectopic activation of skin-like epithelium. Conversely, introduction of PAX6 into skin cells induces many cornea-like features. These studies indicate a critical role of PAX6 for maintaining cornea epithelium, but the mechanisms, which determine cornea vs. skin identity, are still unknown. Our hypothesis is that PAX6 contributes to cornea identity through tissue-specific DNA regulatory elements called enhancers. Enhancers are responsible for coordinating tissue-specific gene expression and for the development of specific cell types. Enhancers are also thought to be of considerable medical importance as genetic variation in and epigenetic regulation of enhancers is thought to play a major role in susceptibility to disease. Despite their biological and medical importance, little is known about the enhancers of the cornea and disease. Advances in chromatin biology and high-throughput sequencing provide new, powerful tools to identify tissue-specific enhancers of the cornea and have the potential to expand our knowledge of cornea disease. Here we propose to (1) identify enhancers, which distinguish cornea from skin epithelium via ChIP-seq; (2) determine how tissue-specific enhancers are regulated in the cornea by chromatin analysis and reporter studies in cultured cells, animal model transplants; and (3) determine the role of PAX6 in the regulation of tissue-specific enhancers. To accomplish these goals, an interdisciplinary approach is needed. The combined expertise and resources of two labs (eye, skin) provides an ideal path to identifying cornea- vs. skin-specific regulation and will result in a better understanding of how ectopic skin features become activated in the cornea. This major effort will result in the identification of genome-wide enhancers in the cornea, critical to stem cell biology and translational studies, and the generation of new genetic reagents to study cornea disease.

描述（由适用提供）：角膜是一种专门的上皮，可保护眼睛并保持正常视力的透明度。角膜或其干细胞（边缘干细胞）的疾病会影响全球数百万的人，并由病因范围造成，范围从遗传缺陷到感染，感染或创伤。视力丧失通常是由于透明的角膜转化为皮肤样上皮的结果。现在，我们实验室和其他人的最新工作表明，由于失去PAX6表达，这种关键过渡可能是宝贵的。 PAX6的下调导致角膜特征和基因表达的丧失以及皮肤样上皮的生态激活。相反，将PAX6引入皮肤细胞会影响许多类似角膜的特征。这些研究表明，PAX6在维持角膜上皮方面的关键作用，但是确定角膜与皮肤身份的机制仍然未知。我们的假设是，PAX6通过组织特异性的DNA调节元件（称为增强子）有助于角膜身份。增强剂负责协调的组织特异性基因表达和特定细胞类型的发展。由于遗传变异的遗传变异，增强剂的表观遗传调节被认为在疾病易感性中起主要作用，因此增强子也被认为具有医学意义。尽管它们具有生物学和医学的重要性，但对角膜和疾病的增强子知之甚少。染色质生物学和高通量测序的进步提供了新的强大工具，可以识别角膜的组织特异性增强子，并有可能扩大我们对角膜疾病的了解。在这里，我们建议（1）识别增强子，这些增强剂通过芯片seq将角膜与皮肤上皮区分开；（2）确定如何通过染色质分析和在培养细胞（动物模型移植）中的染色质分析和报告基因研究来调节组织特异性增强子；（3）确定PAX6在组织特异性增强子调节中的作用。为了实现这些目标，需要一种跨学科的方法。两个实验室（眼，皮肤）的组合专业知识和资源为识别角膜与皮肤特定调节提供了理想的途径，并将更好地了解角膜中生态皮肤特征如何激活。这项重大努力将导致角膜中全基因组增强子的鉴定，对干细胞生物学和翻译研究至关重要，以及生成新的遗传试剂来研究角膜疾病。