Exploring the link between mutant RNA expression and trinucleotide repeat disease

项目摘要/摘要 Fuchs内皮性角膜营养不良(FECD)是最常见的扩张型 重复障碍，影响了4%的美国人口。FECD导致进行性失明 由于早衰和角膜内皮细胞的凋亡。目前，唯一的 治疗FECD的方法是角膜移植。该项目旨在开发一种 在分子水平上了解疾病以指导疾病的发展 治疗。在70%的晚发性FECD中，CTG三核苷酸扩增位于 TCF4基因产生突变的CUG RNA，可在细胞核内的病灶中发现 角膜内皮细胞。这些突变的rna焦点被认为会导致细胞功能障碍。 FECD中的角膜内皮细胞。这个项目试图回答一个问题，即如何 少数几个分子可以对细胞生理学产生巨大影响。第一个目标是 使用质谱仪(MS)鉴定与突变的CUG-重复RNA结合的蛋白质。这个 第二个目标是确定类粘虫蛋白的翻译后修饰 (MBNL)在FECD患者来源的细胞中。MBNL与之前的 研究人员对其CUG重复RNA的倾向。MBNL是一个剪接因子，它的缺失 功能缺陷可以解释FECD中的剪接缺陷。第三个目标是验证 Aim One和Aim One中产生的候选蛋白质和翻译后修饰结果 目标二。 科里实验室的环境是协作的，学员受益于 科里博士的指导，以及几位资深研究人员的协助。这 该项目与德克萨斯大学西南眼科大学的维诺德·穆萨博士合作 部门。本次团契培训有四个主要目标：1) 了解RNA生物学和RNA相关疾病的文献，2)成为 熟练而成功的资助金和手稿作者，3)成为熟练的实验者， 4)在所有听众面前成为一个口齿清晰、引人入胜的演讲者。所获得的技能 通过这些目标将有助于最终实现成为一名首席调查员 在一个研究实验室里。

Project Summary/Abstract Fuchs’ endothelial corneal dystrophy (FECD) is the most common expanded repeat disorder, affecting 4% of the US population. FECD causes progressive blindness due to premature senescence and apoptosis in corneal endothelium. Currently, the only curative treatment for FECD is a corneal transplant. This project aims to develop an understanding of the disease at a molecular level to inform the development of treatments. In 70% of late onset FECD, a CTG trinucleotide expansion in intron 2 of the TCF4 gene produces a mutant CUG RNA which can be found in foci in the nucleus of corneal endothelial cells. These mutant RNA foci are thought to cause cellular dysfunction in FECD corneal endothelial cells. This project seeks to answer the question of how a handful of molecules can have a dramatic impact on cell physiology. The first aim is to identify proteins that bind mutant CUG-repeat RNA using mass spectrometry (MS). The second aim is to identify post-translational modifications on Muscleblind-like protein (MBNL) in a FECD patient-derived cells. MBNL has been implicated by previous researchers for its propensity to CUG repeat RNA. MBNL is a splicing factor and its loss of function may explain splicing defects seen in FECD. The third aim is to validate the candidate proteins and post-translational modification results generated in aim one and aim two. The environment in the Corey laboratory is collaborative and trainees benefit from Dr. Corey’s mentorship and also the assistance of several senior researchers. This project is in collaboration with Dr. Vinod Mootha in the UT Southwestern Ophthalmology Department. There are four principal goals for this fellowship training period: 1) to be knowledgeable of the literature in RNA biology and RNA related diseases, 2) to be a proficient and successful grant and manuscript writer, 3) to be a proficient experimentalist, and 4) to be an articulate and engaging speaker to all audiences. The skills obtained through these goals will aid in the ultimate achievement of becoming a lead investigator in a research laboratory.