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)诊断的问题有许多 对耳科医生的挑战。因为最终标准是 PLF的术前诊断和术中确认尚未得到证实 虽然案件的定义标准已经确定，但仍不确定。之前 临床研究提示，PLF可能比 以前受到赞赏，但特点是缺乏前瞻性 设计和适当的控制组。 这项研究的第一阶段建议利用 外淋巴蛋白开发一种临床适用的标记 对于PLF的术中诊断具有特异性，并且足够敏感 检测低至2-5微升的外淋巴液 受到其他外来流体的污染。 还建议进行第二阶段临床试验： 1)假设开发了一种用于PLF的敏感标记，则多个 机构前瞻性观察性研究预计将研究 可验证的PLF的患病率和识别诊断预测 听力和前庭功能障碍患者的标准。 2)处理有关潜在假阴性结果的不确定性 来自间歇性PLF和微瘘的前瞻性随机研究 涉及前庭功能障碍患者的临床试验 建议对推定的PLF进行手术治疗的疗效比较 并与对照组进行前庭康复治疗。 这项研究的目的是为了了解儿童的发展状况 视网膜细胞实现特化的机制 子类型。 玻璃突变特异地移除了 果蝇的视觉系统；这些细胞开始发育为神经元，但 不能表达光感受器特异的基因，并在以后死亡。玻璃杯 编码一个含有五个锌指结构域的蛋白质；这样的蛋白质已经 在其他生物体中表现为转录因子。因此，玻璃 可能直接作用于调节感光细胞特异性基因 表情。我们已经证明玻璃蛋白在体外结合到 一个这样的基因(视紫红质)的增强子元件的序列。玻璃杯 在发育中的视网膜的所有细胞中都有表达，但只有 活跃在发育中的光感受器(通过显示对 玻璃DNA结合位点的报告基因)。因此，对玻璃的监管 活性是光感受器细胞发育的关键，这发生在 在两个层面上：转录时和翻译后(通过输入 位置信号)。为了推行这一双重监管，我们将实施两项 项目： 1)为了了解玻璃转录调控，我们将进行 玻璃基因启动子突变的功能分析。 2)为了识别与玻璃相互作用的新基因，我们将使用一系列 第一个将是针对显性增强子和 弱玻璃的抑制者。初步筛查成功 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)。一旦被孤立，我们将描述我们小说的影响 突变，并从长远来看，克隆负责研究的基因 它们的分子功能。