Functional Analysis of Optineurin in Glaucoma

Optineurin 在青光眼中的功能分析

• 批准号: 6774271
• 负责人: Mansoor Sarfarazi
• 金额: $ 31.33万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6774271
• 关键词: clinical research; developmental genetics; developmental neurobiology; disease /disorder etiology; gene expression; gene mutation; genetic screening; genetic susceptibility; genotype; glaucoma; human subject; immunocytochemistry; in situ hybridization; laboratory mouse; messenger RNA; patient oriented research; phenotype; protein localization; protein protein interaction; protein quantitation /detection; protein structure function; transmission electron microscopy; yeast two hybrid system

DESCRIPTION (provided by applicant): Glaucoma is an optic neuropathy that affects over 67 million people worldwide. This condition has broad clinical subtypes ranging from birth to very late in life. The most common form, Primary Open Angle Glaucoma (POAG) has a prevalence of about 1% of white population over 40 years of age. Several genetic loci have been identified for POAG but so far only mutations in Myocilin gene are reported in juvenile-onset and certain other adult-onset cases. Recently, we identified a gene that is primarily involved in a subgroup of Adult-Onset POAG, commonly known as Normal Tension Glaucoma (NTG). This gene that we named Optineurin (OPTN) is mutated in 16.7% of our hereditary NTG families, maps to the GLC1E locus on 10p14, has 13 coding exons and encodes for a 577-amino acids protein (~66-kDa). Our OPTN protein studies showed co-localization with Golgi, secretion into aqueous humor and expression in many ocular and non-ocular tissues. Cloning of both mouse and monkey genes showed very similar patterns of mRNA and protein expression to human OPTN. It has also been shown by other investigators that OPTN interacts with Ad E3-14.7K, Huntingtin, TFIIIA, RAB8 and 2 other unknown kinases. Although existing evidence suggest that OPTN through its interaction with other proteins may be utilizing TNF-a or Fas-Ligand pathways to mediate apoptosis, inflammation or vasoconstriction but, as yet, these is no clear indication on how OPTN mutations lead to either NTG or POAG. Therefore, as an initial step towards understanding the ocular function of this gene and its protein products we propose the followings. 1)-Screen a large number of glaucoma patients for OPTN mutations in order to establish a genotype/phenotype correlation; 2)-Use OPTN as "bait" and search for new interacting proteins by Yeast Two Hybrid System and to identify motifs that are important for OPTN function; 3)-Determine ultracellular localization of OPTN in normal and glaucomatous eyes by immunogold labeling; 4)-Study OPTN distribution in normal and glaucomatous eyes by immunohistochemistry and to determine its potential differential tissue expression patterns; and 5)-Use In Situ hybridization to study developmental expression patterns of Optn in mouse embryos. At the conclusion of this investigation, it is anticipated that our study will provide essential scientific information that one day may contribute to the design of innovative and futuristic drug intervention for this group of optic neuropathies.

描述(申请人提供)：青光眼是一种视神经病变，全世界有超过6700万人受到影响。这种疾病有广泛的临床亚型，从出生到晚年不等。最常见的形式是原发性开角型青光眼(POAG)，在40岁以上的白人人口中约有1%的患病率。POAG有几个基因位点已被确定，但到目前为止，只有在青少年发病和某些其他成人发病病例中报道了myocin基因突变。最近，我们发现了一个基因，该基因主要与成人型POAG的一个亚群有关，通常被称为正常眼压性青光眼(NTG)。我们命名为Optineurin(OPTN)的这个基因在16.7%的遗传性NTG家族中发生突变，定位于10p14的GLC1E基因座，有13个编码外显子，编码577个氨基酸的蛋白质(~66 kDa)。我们的OPTN蛋白研究表明，OPTN蛋白与高尔基体共定位，分泌到房水中，并在许多眼和非眼组织中表达。克隆小鼠和猴子的基因显示出与人类OPTN非常相似的mRNA和蛋白质表达模式。其他研究人员也发现，OPTN与AdE3-14.7K、Huntingtin、TFIIIA、Rab8等2种未知蛋白激酶相互作用。虽然现有的证据表明，OPTN通过与其他蛋白质的相互作用，可能利用肿瘤坏死因子-a或Fas-配体途径来介导细胞凋亡、炎症或血管收缩，但目前还没有明确的迹象表明OPTN是如何突变的 通向NTG或POAG。因此，作为了解该基因及其蛋白产物的眼功能的第一步，我们提出如下建议。1)-筛选大量青光眼患者的OPTN突变，以建立基因/表型相关性；2)以OPTN为“诱饵”，通过酵母双杂交系统寻找新的相互作用蛋白，并确定对OPTN功能至关重要的基序；3)通过免疫金标记法确定OPTN在正常和青光眼眼中的超细胞定位；4)通过免疫组织化学方法研究OPTN在正常和青光眼眼中的分布并确定其潜在的差异组织表达模式；以及5)使用原位杂交技术研究OPTN在小鼠胚胎中的发育表达模式。在本次调查结束时，预计我们的研究将提供基本的科学信息，有朝一日可能有助于为这一组视神经疾病设计创新和未来的药物干预措施。