DEVELOPMENT OF CLINICAL MARKER FOR PERILYMPH FISTULA

外淋巴瘘临床标志物的开发

• 批准号: 3550481
• 负责人: STEVEN A TELIAN
• 金额: $ 26.1万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-15 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3550481
• 关键词: audiometry; biomarker; cerebrospinal fluid; clinical trials; cochlea; ear disorder; ear disorder diagnosis; ear surgery; guinea pigs; hearing disorders; high performance liquid chromatography; human subject; human therapy evaluation; human tissue; ion exchange chromatography; labyrinth; labyrinth disorder; labyrinthectomy; mathematical model; middle ear; model design /development; monoclonal antibody; perilymph; postmortem; protein purification; protein sequence; synthetic peptide; western blottings

Issues surrounding the diagnosis of perilymph fistula (PLF) present many challenges to the otologist. Since definitive criteria for the preoperative diagnosis and intraoperative confirmation of PLF have not been established, case definition criteria are uncertain. Prior clinical studies raise the suggestion that PLF may be more common than previously appreciated, but are characterized by a lack of prospective design and appropriate control groups. Phase I of this study proposes to utilize the unique constituency of perilymph proteins to develop a clinically applicable marker that is specific for the intraoperative diagnosis of PLF, and sensitive enough to detect perilymph in quantities as low as 2-5 microliters despite contamination by other adventitious fluids. Phase II clinical trials are also proposed: 1) assuming that a sensitive marker for PLF is developed, a multi- institutional prospective observational study is anticipated to study the prevalence of verifiable PLF and identify diagnostic prediction criteria in patients with auditory and vestibular dysfunction. 2) to address uncertainties regarding potential false negative results from intermittent PLF and microfistulae, a prospective randomized clinical trial involving patients with vestibular dysfunction is proposed to compare the efficacy of surgical treatment for presumed PLF with a control group treated in vestibular rehabilitation therapy. The aim of the research proposed is to understand the developmental mechanisms by which the cells of the retina achieve their specialized sub-types. glass mutations specifically remove the photoreceptor neurons of the Drosophila visual system; these cells begin to develop as neurons but fail to express photoreceptor specific genes, and later die. glass encodes a protein with five Zinc-finger domains; such proteins have been showm in other organisms to act as transcription factors. Thus glass may act directly to regulate photoreceptor cell-specific gene expression. We have shown that the glass protein binds in-vitro to sequences from enhancer elements of one such gene (a rhodopsin). glass is expressed in all the cells of the developing retina, but it is only active in the developing photoreceptors (by showing the affect on a reporter gene of glass DNA binding sites). Thus the regulation of glass activity is critical for photoreceptor cell development and this occurs at two levels: at transcription, and after translation (by incoming positional signals). To pursue this dual regulation we will conduct two projects: 1) To understand glass transcriptional regulation we will undertake a functional analysis of the glass gene promoter by mutational studies. 2) To identify novel genes that interact with glass we will use a series of genetic screens and the first will be for dominant enhancers and suppressors of weak glass areles. Preliminary screens have successfully tested the feasibility of this approach (we have recovered three such mutations). Once isolated we will characterize the effects of our novel mutations and in the long term, clone the genes responsible to study their molecular functions.

围绕外淋巴瘘（PLF）的诊断存在许多问题 对耳科医生的挑战。 由于确定的标准 术前诊断和术中确认PLF尚未 已经确定，病例定义标准尚不确定。 事先的 临床研究表明，PLF 可能比 以前受到赞赏，但其特点是缺乏前瞻性 设计和适当的对照组。 本研究的第一阶段建议利用独特的选区 外淋巴蛋白开发临床适用的标记物 对PLF术中诊断具有特异性，且足够灵敏 检测外淋巴量低至 2-5 微升，尽管 其他外来液体的污染。 还提出了 II 期临床试验： 1) 假设开发了 PLF 敏感标记， 机构前瞻性观察研究预计将研究 可验证的 PLF 患病率并确定诊断预测 听觉和前庭功能障碍患者的标准。 2) 解决潜在假阴性结果的不确定性 间歇性 PLF 和微瘘，一项前瞻性随机 涉及前庭功能障碍患者的临床试验是 建议比较手术治疗对假定的 PLF 的疗效 对照组接受前庭康复治疗。 拟议研究的目的是了解发育 视网膜细胞实现其特化的机制 子类型。 玻璃突变专门去除了感光神经元 果蝇视觉系统；这些细胞开始发育为神经元，但是 无法表达光感受器特异性基因，然后死亡。玻璃 编码具有五个锌指结构域的蛋白质；此类蛋白质已被 在其他生物体中显示出作为转录因子的作用。 因此玻璃 可能直接调节感光细胞特异性基因 表达。 我们已经证明玻璃蛋白在体外结合 来自此类基因（视紫红质）的增强子元件的序列。玻璃 在发育中的视网膜的所有细胞中表达，但仅 活跃于发育中的光感受器（通过显示对光感受器的影响） 玻璃 DNA 结合位点的报告基因）。 因此玻璃的调节 活性对于感光细胞的发育至关重要，并且这种情况发生 在两个层面上：转录时和翻译后（通过传入 位置信号）。 为了实现这种双重监管，我们将进行两项 项目： 1）为了了解玻璃转录调控，我们将进行 通过突变研究对玻璃基因启动子进行功能分析。 2）为了识别与玻璃相互作用的新基因，我们将使用一系列 遗传筛选，第一个将是显性增强子和 弱玻璃区域的抑制剂。 初步筛选已成功 测试了这种方法的可行性（我们已经恢复了三个这样的 突变）。 一旦孤立，我们将描述我们小说的效果 突变，从长远来看，克隆负责研究的基因 它们的分子功能。