MOLECULAR GENETIC STUDIES IN CORNEAL DYSTROPHIES

角膜营养不良的分子遗传学研究

• 批准号: 2668357
• 负责人: Richard W Yee
• 金额: $ 15万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2668357
• 关键词: artificial chromosomes; autosomal dominant trait; blood chemistry; clinical research; complementary DNA; cornea disorder; family genetics; gene mutation; genetic mapping; genetic markers; genetic polymorphism; genome; human genetic material tag; human subject; linkage mapping; lymphoblast; molecular cloning; molecular genetics; nucleic acid hybridization; polymerase chain reaction

The general objectives of this proposal are to gain an education and basic laboratory training in molecular genetic methods and human genetics, in general, and to apply these skills to ophthalmologic disorders. The purpose is to contribute to the understanding and treatment of inherited ocular disorders, with a special emphasis on corneal diseases. These objectives will be met by attending graduate level courses and conferences as described (Appendix B-1) and by performing gene linkage studies in families with Reis Bucklers Corneal Dystrophy (RBCD). The RBCD proposal is the platform for my training to be an independent physician-scientist in the field of molecular genetics. RBCD is a dominantly inherited corneal disorder, has an infantile onset, complete penetrance and variable expressivity, and affects primarily central vision causing <20/200 visual acuity, disabling photophobia, and sharp stabbing pain from recurrent erosions in severely affected individuals. The slit lamp appearance of the milder cases closely resembles the reticular appearance of post EXCIMER laser corneal haze. A basic molecular understanding of RBCD may shed light on the more common problem of corneal scarring, the most common cause for corneal transplantation in the American elderly. Once the RBCD gene is cloned and characterized, the knowledge gained about the pathophysiologic mechanisms may lead to a greater understanding of corneal wound healing, scarring and recurrent erosion syndrome. Briefly, the following techniques are employed to perform gene-linkage analysis. First, the pedigree is ascertained and blood is collected from significant family members for DNA preparation. If needed, lymphoblast cell lines are established. Then, using highly informative microsatellite DNA markers, the pedigree is examined. Linkage analysis is performed in an effort to establish the chromosomal location of the gene for RBCD. Once linkage to a chromosome is found, fine structure linkage mapping, in conjunction with physical mapping, will be used to narrow down the region containing the RBCD locus. Yeast artificial chromosome (YAC) contigs which span this region will be constructed (or identified). A number of techniques such as exon trapping (Duyk 1990, Bucklers 1991), solution hybridization (Hoffman 1987),and detection of Hpall tiny fragment (HTF) islands (Conneally 1984) will then be used to identify expressed sequences from the RBCD region. Once the RBCD gene is cloned and characterized, understanding the pathophysiologic process of this anterior basement membrane disorder will be helpful in illuminating mechanisms in other corneal related conditions. In addition, this knowledge will assist in rational planning of diagnostic and counseling services as well as preventative therapies and treatment for affected patients.

这项建议的总目标是获得教育， 分子遗传学方法和人类遗传学的基本实验室培训 遗传学，一般来说，并将这些技能应用于眼科 紊乱其目的是促进理解和 治疗遗传性眼部疾病，特别强调 角膜疾病这些目标将通过参加研究生 等级课程和会议（附录B-1）， 在Reis Bucklers角膜的家族中进行基因连锁研究 营养不良（RBCD）。区域局的建议是我接受培训的平台， 成为分子遗传学领域的独立医生科学家。 RBCD是一种显性遗传性角膜疾病，具有婴儿发病， 完全的恍惚和可变的表达，并影响主要是 中心视力，导致视力<20/200，致残性恐惧症，和 在严重受影响的患者中， 个体裂隙灯外观较温和的情况下密切 类似于EXCIMER激光后角膜混浊的网状外观。 对RBCD的基本分子理解可能有助于了解更常见的 角膜瘢痕形成的问题，最常见的原因是角膜 移植在美国老年人。一旦RBCD基因被克隆， 表征，获得的关于病理生理机制的知识 可能有助于更好地了解角膜伤口愈合、瘢痕形成 和复发性糜烂综合征。 简言之，采用以下技术进行基因连锁 分析.首先，确定谱系，并从 重要的家庭成员进行DNA制备。如果需要，淋巴母细胞 建立细胞系。然后，利用高信息量的微卫星 DNA标记，系谱检查。连锁分析在 努力确定RBCD基因的染色体定位。 一旦发现与染色体的连锁，精细结构连锁作图， 结合物理映射，将用于缩小 包含RBCD位点的区域。酵母人工染色体 将构建（或鉴定）跨越该区域的重叠群。一 许多技术，如外显子捕获（Duyk 1990，Bucklers 1991）， 溶液杂交（霍夫曼1987）和检测Hpall微小 碎片（HTF）岛（Conneally 1984）将用于识别 来自RBCD区域的表达序列。 一旦RBCD基因被克隆和鉴定， 这种前基底膜疾病的病理生理过程将 有助于阐明其他角膜相关的机制 条件此外，这些知识将有助于合理规划 诊断和咨询服务以及预防性治疗 以及对受影响患者的治疗。

项目成果

A novel mutation of the Keratin 12 gene responsible for a severe phenotype of Meesmann's corneal dystrophy.

角蛋白 12 基因的一种新突变导致了 Meesmann 角膜营养不良的严重表型。

• 发表时间: 2007
• 期刊: Molecular vision
• 影响因子: 2.2
• 作者: Sullivan,LoriS;Baylin,EricB;Font,Ramon;Daiger,StephenP;Pepose,JayS;Clinch,ThomasE;Nakamura,Hisashi;Zhao,XinpingC;Yee,RichardW
• 通讯作者: Yee,RichardW