Molecular characterization of congenital cataract

先天性白内障的分子特征

• 批准号: 8582345
• 负责人: Elena V Semina
• 金额: $ 22.5万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8582345
• 关键词: Adolescent; Affect; Age; Alleles; Biology; Birth; Blindness; Candidate Disease Gene; Cataract; Childhood; Classification; Counseling; Crystallins; Cytoskeletal Proteins; Data; Development; Diagnosis; Differentiation and Growth; Disease; Evaluation; Eye Development; Family; Family member; Frequencies; Gene Mutation; Genes; Genetic; Genetic screening method; Human; Knowledge; Lead; Lens Opacities; Membrane Proteins; Molecular; Molecular Diagnosis; Mutation; Patients; Phenotype; Recurrence; Reporting; Risk; Role; Sequence Analysis; Severities; Testing; Variant; Zebrafish; cohort; congenital cataract; exome; exome sequencing; genetic pedigree; improved; lens; mutant; new technology; novel; novel diagnostics; patient population; proband; public health relevance; research study; vision development

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

首席研究员/项目总监（最后、第一、中间）：Semina, Elena V. 描述：说明该应用程序的广泛、长期目标和具体目标，并参考该项目的健康相关性。描述 简明地阐述了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 与应用程序分开时，旨在作为对拟议工作的简洁而准确的描述。如果申请获得资助，这 描述将按原样成为公共信息。因此，请勿包含专有/机密信息。不要超出空间 假如。 每 10,000 名新生儿中就有 1-15 人患有儿童白内障。先天性白内障是最严重的 由于干扰正常视觉发育和永久性视力障碍的可能性而处于光谱末端 失明。遗传学研究已经发现许多与各种形式的疾病相关的基因突变。 白内障。先天性/青少年先天性疾病患者中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）是一种新技术，可用于识别已知和已知的致病突变。 新基因，因此可以显着促进儿童白内障的表征/分类 分子水平。在初步实验中，WES 用于筛选 22 个受影响家系的先证者 患有家族性显性先天性/青少年白内障，并在已知白内障中发现了致病突变 基因和一种新的晶状体蛋白基因存在于 45% (10/22) 的家族中。在本提案中，我们重点关注 通过分析鉴定与人类先天性/青少年白内障有关的新遗传因素 家族病例。第一个目标是确定之前十二种与白内障相关的基因突变 确定了已知白内障基因没有突变的家族和另外十二个受影响的家系。这 第一步是对其他先证者进行全外显子组测序/已知白内障基因分析，即 可能会识别出 40-50% 家庭的致病突变。家族中新遗传因素的发现 最初将通过评估在脊椎动物中具有已知作用的基因来接近无突变 晶状体发育/功能或位于先前报道的白内障位点内。接下来，WES 额外 突变阴性家系中的家庭成员及其外显子组变异的综合分析将 旨在识别新因素中的有害变异。第二个目的是验证 先天性/幼年性白内障的新因素，通过对新的候选白内障因素进行功能分析 斑马鱼和对一大群患有白内障和相关眼病的人类患者进行检查 这些基因突变的条件。与儿科相关的基因/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开发和利用 基因检测可准确诊断并预测表型严重程度和复发风险，从而提供 为受影响的家庭提供优质咨询。除此之外，对遗传机制的了解 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。 绩效站点（组织、城市、州） 儿科 儿童研究所 威斯康星医学院 8701水城栈道 密尔沃基, WI 53226 关键人员。请参阅说明。根据需要使用延续页面以如下所示的格式提供所需信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，姓氏在前。 名称 组织在项目中的角色 Elena V. Semina，博士 威斯康星医学院 PI 披露许可声明。仅适用于 SBIR/STTR。请参阅说明。是 否 PHS 398（修订版 05/01）第 _2___ 表格第 2 页