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

主要研究者/项目负责人（最后，第一，中间）：Semina，Elena V。 说明：说明申请的广泛、长期目标和具体目标，并参考项目的健康相关性。描述 简要介绍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。这个抽象 是为了作为一个简洁和准确的描述拟议的工作时，从申请分开。如果申请得到资助， 这样的描述将成为公开信息。因此，不包括专有/机密信息。不要超过空间 提供了 每10 000名新生儿中就有1-15人患有儿童白内障。先天性白内障最严重 由于干扰正常的视觉发育和永久性的可能性， 失明遗传研究已经确定了与各种形式的遗传病相关的许多基因的突变。 白内障这些已知基因在先天性/青少年性乳腺癌患者中的确切突变频率 白内障是未知的，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，因此可以显着促进儿童白内障的表征/分类， 分子水平。在初步实验中，WES被用来筛选22个家系的先证者， 患有家族性显性先天性/青少年白内障，并在已知白内障中确定了致病突变 45%（10/22）的家系中发现了一个新的晶状体蛋白基因。在本提案中，我们重点关注 通过分析人类先天性/青少年性白内障新的遗传因素 家族案件。第一个目的是在先前的12个白内障患者中鉴定与白内障相关的基因突变。 确定了已知白内障基因中没有突变的家族和另外12个受影响的家系。的 第一步将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 可能在40-50%的家庭中发现致病突变。家族中新遗传因子的发现 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进行。 透镜发育/功能或位于先前报道的白内障位点内。接下来，WES的附加 突变阴性家系中的家庭成员及其外显子组变异的联合分析将是 以确定新因素中的有害变体。第二个目的是核实参与 先天性/青少年白内障的新因素，通过对新候选白内障因素的功能分析， 斑马鱼和受白内障和相关眼部疾病影响的更大人群的检查 这些基因突变的条件。与儿科疾病相关的基因/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白内障的开发和利用， 基因检测用于准确诊断和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上级咨询服务。除此之外，了解遗传机制， 白内障将提高我们对透镜生物学的认识， 所有年龄段的透镜混浊。 履约地点（组织、城市、州） 儿科 儿童研究所 威斯康星州医学院 沃特敦普兰克路8701号 密尔沃基，威斯康星州53226 关键人员。参见说明。根据需要使用续页以如下所示的格式提供所需信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，姓在前。 名称组织在项目中的角色 Elena V. Semina，博士威斯康星州医学院PI 披露许可声明。仅适用于SBIR/STTR。参见说明。是否 PHS 398（版本05/01）第_2页_表格第2页