AMINOGLYCOSIDE SUPPRESSION OF PREMATURE STOP MUTATIONS IN CFTR

许多人类疾病是由过早终止翻译引起的。 突变。由于这类突变的流行，一种 防止在提前停止时终止翻译的机制 突变将有可能减少或消除这种疾病 一群患有多种遗传病的个体， 包括癌症，糖尿病，泰·萨克斯，β-地中海贫血， 高胆固醇血症，以及其他许多疾病。另一个例子是这种疾病 囊性纤维化(CF)。Cf是由编码基因突变引起的。 CF跨膜电导调节器(CFTR)。而最多的 CFTR基因的常见突变是苯丙氨酸缺失 残留物508(Delta F508)，大约10%的CF患者携带 至少有一次过早停止突变。在初步研究中，我们获得了 有证据表明，在CF患者中发现的过早停止突变可能是 被氨基糖苷类抗生素抑制，导致表达 全长、功能性CFTR。这些结果表明，临床上 采用氨基糖苷类药物治疗可能恢复CFTR功能 Cf患有这类突变的患者。然而，重要的是 我们首先更详细地了解翻译的机制 终止以及氨基糖苷类如何颠覆这一过程。至 探讨以氨基糖苷类药物为基础的治疗方法的可行性 抑制致病的过早停止突变，我们建议 以下是具体目标： 特异性目的1：研究氨基糖苷类药物对血管内皮细胞生长的抑制作用 Cftr基因过早停止突变。 特异性目标2：研究氨基糖苷类化合物抑制A细胞的能力 人类CFTR基因自然发生的过早停止突变 在转基因小鼠模型中表达。 具体目标3：确定序列上下文如何影响效率 在人类细胞中的翻译终止。 具体目标4：描述翻译终止的机制 并研究氨基糖苷类化合物如何改变哺乳动物的这一过程 细胞。

Many human diseases are caused by premature translation termination mutations. Because of the prevalence of this class of mutation, a mechanism that prevented translation termination at premature stop mutations would have the potential to reduce or eliminate the disease of a subset of individuals afflicted with a wide range of genetic diseases, including cancer, diabetes, Tay Sachs, beta-thalassemia, hypercholesterolemia, and many others. Another example is the disease cystic fibrosis (CF). CF is caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR). While the most common mutation in the CFTR gene is the deletion of a phenylalanine at residue 508 (delta F508), roughly 10 percent of all CF patients carry at least one premature stop mutation. In preliminary studies, we obtained evidence that premature stop mutations found in CF patients can be suppressed by aminoglycoside antibiotics, resulting in the expression of full-length, functional CFTR. These results suggest that a clinical strategy utilizing aminoglycoside treatment may restore CFTR function in CF patients with this class of mutation. However, it is important that we first understand in greater detail the mechanism of translation termination and how aminoglycosides act to subvert this process. To explore the feasibility of using aminoglycoside-based therapy to suppress disease-causing premature stop mutations, we propose the following Specific Aims: SPECIFIC AIM 1: Characterize the aminoglycoside-mediated suppression of premature stop mutations in the CFTR gene. SPECIFIC AIM 2: Investigate the ability of aminoglycosides to suppress a naturally-occurring premature stop mutation in the human CFTR gene expressed in a transgenic mouse model. SPECIFIC AIM 3: Determine how sequence context affects the efficiency of translation termination in human cells. SPECIFIC AIM 4: Characterize the mechanism of translation termination and investigate how aminoglycosides alter this process in mammalian cells.