Drug Screening Assays for Charcot-Marie-Tooth Disease

描述（由申请人提供）：遗传性周围神经病（也称为遗传性运动感觉神经病，HMSN）是影响神经系统的最常见遗传性疾病之一。最温和形式的人类周围神经病，腓骨肌萎缩症（CMT），在受影响的个体中引起运动和感觉神经的进行性恶化、肌肉萎缩和慢性疼痛/疲劳。大多数遗传性外周髓鞘病是由一个关键的髓鞘基因，外周髓鞘蛋白22（PMP 22），这是分类为CMT 1A的重复。因此，治疗这种遗传性髓鞘病的最直接的途径之一是实现相对微妙（<2倍）的变化。 在基因调控中。最近使用候选化合物降低PMP 22表达水平的原理验证研究在CMT 1A的啮齿动物模型中显示出有益效果。然而，这些药剂尚未在临床试验中显示出有效性，因此关键在于开发可用于鉴定可实现治疗性降低PMP 22水平的化合物的有效筛选工具。我们最近对PMP 22调控的研究已经阐明了该基因中新的调控元件，本提案的目标是设计和功能性地 验证新的检测方法，以确定减少PMP 22表达的小分子。我们建议利用一种新的技术，涉及自定义锌指核酸酶，这将使我们能够嵌入一系列的orthopathic记者在本地Pmp 22位点。这些检测试剂盒将创建一系列互补检测试剂盒，用于一级和二级筛选检测试剂盒。提出了一系列验证试验，以确定报告基因试验是否受到适当监管，这些试验将适用于NIH化学基因组学中心的高通量筛选。 公共卫生相关性：影响神经系统的最常见的遗传性疾病之一是腓骨肌萎缩症，大约每3000人中就有1人患病。这种疾病最常见的原因是PMP 22基因的重复（导致CMT 1A），但尚未开发出有效的治疗方法来防止周围神经保护性髓鞘的进行性恶化。该提案旨在创建用于小分子筛选的新工具，旨在开发CMT 1A的新治疗方法。 免责声明：请注意，以下评论是由审查员在研究部分会议之前准备的，并且以基本上未经编辑的形式提供。 虽然审查人员有机会根据小组讨论情况更新或修订其书面评价，但不能保证在会议讨论之后更新了个人评论。 因此，这些评论可能并不完全反映小组讨论结束时个人评论员的最终意见或小组的最终多数意见。因此，讨论的简历和摘要是评审员在会议上实际认为关键的最后一句话。

DESCRIPTION (provided by applicant): Hereditary peripheral neuropathies (also known as hereditary motor sensory neuropathies, HMSN) are among the most common genetic diseases affecting the nervous system. The mildest form of human peripheral neuropathy, Charcot-Marie- Tooth (CMT) disease, causes progressive deterioration of both motor and sensory nerves, muscular atrophy, and chronic pain/fatigue in affected individuals. A majority of inherited peripheral myelinopathies are caused by duplication of a critical myelin gene, Peripheral Myelin Protein 22 (PMP22), which is classified as CMT1A. Therefore, one of the most straightforward avenues for treating this inherited myelinopathy is to achieve a relatively subtle (<2-fold) change in gene regulation. Recent proof-of-principle studies using candidate compounds to reduce PMP22 expression levels have shown beneficial effects in rodent models of CMT1A. However, these agents have not yet been shown to be effective in clinical trials, and therefore it is criticl to develop effective screening tools that can be used to identify compounds that can achieve a therapeutic reduction in PMP22 levels. Our recent studies of PMP22 regulation have elucidated novel regulatory elements in this gene, and the goal of this proposal is to design and functionally validate novel assays to identify small molecules that reduce PMP22 expression. We propose to utilize a novel technology involving custom zinc finger nucleases, which will allow us to embed a series of orthologous reporters in the native Pmp22 locus. The assays will create a series of complementary assays for use in both primary and secondary screening assays. A series of validation assays are proposed to determine if the reporter assays are appropriately regulated, and such assays will be adapted for high throughput screening at the NIH Chemical Genomics Center. PUBLIC HEALTH RELEVANCE: Relevance One of the most common inherited diseases affecting the nervous system is Charcot-Marie-Tooth disease, affecting approximately 1 in 3000 individuals. The most common cause of this disease is a duplication of the PMP22 gene (resulting in CMT1A), but no effective therapies have yet been developed to prevent the progressive deterioration the protective myelin sheath of peripheral nerves. This proposal is designed to create novel tools for small molecule screening aimed at developing novel treatments for CMT1A. Disclaimer: Please note that the following critiques were prepared by the reviewers prior to the Study Section meeting and are provided in an essentially unedited form. While there is opportunity for the reviewers to update or revise their written evaluation, based upon the group's discussion, there is no guarantee that individual critiques have been updated subsequent to the discussion at the meeting. Therefore, the critiques may not fully reflect the final opinions of th individual reviewers at the close of group discussion or the final majority opinion of the group. Thus the Resume and Summary of Discussion is the final word on what the reviewers actually considered critical at the meeting.