Molecular characterization of congenital cataract

首席研究员/项目主任(最后、第一、中间)：塞米纳、埃琳娜·V。 说明：说明应用程序的广泛、长期目标和具体目标，并参考该项目与健康相关的内容。描述 简明扼要地介绍实现这些目标的研究设计和方法。避免总结过去的成就和使用第一人称。这篇摘要 是为了在脱离应用程序时对提议的工作进行简洁而准确的描述。如果应用程序得到资助，则此 原样的描述将成为公开信息。因此，不包括专有/机密信息。不要超过空格 但前提是。 每10,000名新生儿中有1-15人患上儿童白内障。先天性白内障是最严重的 由于干扰正常的视觉发育和永久性的可能性而导致的光谱结束 失明。遗传学研究已经确定了与各种形式的 白内障。先天/青少年患者这些已知基因突变的确切频率 白内障尚不清楚，许多病例仍在等待分子诊断。全外显子组测序 (WES)是一种新的技术，可用于鉴定已知和 新的基因，因此可以显著促进儿童白内障的特征/分类 分子水平。在初步实验中，使用WES从22个受影响的家系中筛选出先证者 家族性显性先天性/幼年性白内障和已知白内障的致病突变 45%(10/22)的家系携带有新的晶体蛋白基因和1个新的晶体蛋白基因。在这份提案中，我们重点关注 分析人类先天性/幼年性白内障新的遗传因素 家族性病例。第一个目标是在之前的12个项目中确定与白内障相关的基因突变 确定了在已知白内障基因中没有突变的家庭和另外12个受影响的家系。这个 第一步是对额外的先证者进行完整的外显子组测序/已知的白内障基因分析，这是 可能在40%-50%的家庭中发现致病突变。家系中新的遗传因素的发现 如果没有突变，最初将通过评估在脊椎动物中具有已知作用的基因来实现 晶状体发育/功能或位于先前报道的白内障部位内。接下来，WES of Additional 突变阴性家系的家庭成员及其外显子变异的联合分析将是 致力于识别新因素中的有害变种。第二个目的是核实 先天性/青少年白内障新候选因素的功能分析 斑马鱼和对更多患有白内障和相关眼病的人类患者的检查 这些基因突变的条件。儿科疾病相关基因/突变等位基因的鉴定 白内障及其对疾病的具体贡献的确定将指导白内障的开发和利用 用于准确诊断和预测表型严重性和复发风险的基因测试，从而提供 为受影响的家庭提供优质的咨询服务。除此之外，对糖尿病的遗传机制的理解 白内障将提高我们对晶状体生物学的了解，并导致更好的治疗和管理 所有年龄段的晶状体混浊。 演出现场(S)(组织、市、州) 儿科 儿童研究所 威斯康星医学院 水城普兰克路8701号 密尔沃基，威斯康星州53226 关键人员。请参阅说明。根据需要使用续页，以如下所示的格式提供所需信息。 从首席调查员开始。按字母顺序列出所有其他关键人员，姓氏在前。 命名项目中的组织角色 埃琳娜·V·塞米纳，威斯康星医学院博士 披露许可声明。仅适用于SBIR/STTR。请参阅说明。是，不是 小灵通398(05/01版)第_2页_表第2页

Principal Investigator/Program Director (Last, first, middle): Semina, Elena V. DESCRIPTION: State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project. Describe concisely the research design and methods for achieving these goals. Avoid summaries of past accomplishments and the use of the first person. This abstract is meant to serve as a succinct and accurate description of the proposed work when separated from the application. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. Pediatric cataracts are observed in 1-15 per 10,000 births. Congenital cataract represents the most severe end of the spectrum due to interference with normal visual development and the possibility of permanent blindness. Genetic studies have identified mutations in numerous genes associated with various forms of cataracts. The exact frequencies of mutations in these known genes in patients with congenital/juvenile cataract are not known and many cases are still awaiting molecular diagnosis. Whole exome sequencing (WES) is a novel technology that can be utilized for identification of causative mutations in both known and novel genes and thus could significantly facilitate characterization/classification of pediatric cataracts at a molecular level. In preliminary experiments, WES was used to screen probands from 22 pedigrees affected with familial dominant congenital/juvenile cataract and identified causative mutations in known cataract genes and one novel crystallin gene in 45% (10/22) of families. In this proposal, we focus on the identification of novel genetic factors involved in human congenital/juvenile cataract through analysis of familial cases. The first aim is to identify genetic mutations associated with cataract in the twelve previously identified families without mutations in known cataract genes and twelve additional affected pedigrees. The first step will be whole exome sequencing/known cataract gene analysis of the additional probands, which is likely to identify causative mutations in 40-50% of the families. Discovery of novel genetic factors in families without mutations will initially be approached by evaluation of genes which have a known role in vertebrate lens development/function or are located within a previously reported cataract loci. Next, WES of additional family members in mutation-negative pedigrees and combined analysis of their exome variation will be undertaken to identify deleterious variants in novel factors. The second aim is to verify the involvement of novel factors in congenital/juvenile cataract by functional analysis of the new candidate cataract factors in zebrafish and examination of a larger group of human patients affected with cataract and related ocular conditions for mutations in these genes. Identification of the genes/mutant alleles associated with pediatric cataract and determination of their specific contributions to disease will guide development and utilization of genetic tests for accurate diagnosis and predictions of phenotypic severity and recurrence risks, thus offering superior counseling to affected families. In addition to this, understanding of the genetic mechanisms of cataracts will improve our knowledge about lens biology and lead to better treatment and management of lens opacities at all ages. PERFORMANCE SITE(S) (organization, city, state) Department of Pediatrics Children's Research Institute Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226 KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name Organization Role on Project Elena V. Semina, PhD Medical College of Wisconsin PI Disclosure Permission Statement. Applicable to SBIR/STTR Only. See instructions. Yes No PHS 398 (Rev. 05/01) Page _2___ Form Page 2