Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis
Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis
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DOI：
10.1164/ajrccm.163.7.2004001
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发表时间：
2001-06-01
影响因子：
24.7
通讯作者：
Bedwell, DM
中科院分区：
文献类型：
--
作者：
Clancy, JP;Bobök, Z;Bedwell, DM
在这里，我们报告庆大霉素治疗对囊性纤维化跨膜调节因子（CFTR）的生产和功能的CF气道细胞和CF患者与过早停止突变的影响。使用免疫细胞化学和功能[6-甲氧基-N-（3-磺丙基）喹啉（SPQ）为基础的]技术，离体暴露的气道细胞从停止突变CF患者导致识别表面定位CFTR的剂量依赖性的方式。接下来，5名具有终止突变的CF患者和5名CF对照受试者用肠胃外庆大霉素治疗1周，并进行CFTR功能的重复体内测量（鼻电位差[PD]测量和汗液氯化物[Cl-]测试）。治疗期间，与对照组相比，停止突变患者组中Cl-分泌方向的鼻PD读数增加约3倍（p < 0.001），5例CF停止突变患者中有4例在庆大霉素治疗期间至少有一个阅读，Cl-分泌反应超过-5 mV（超极化）。在任何CF对照受试者中均未观察到这种程度的反应（p < 0.05）。在一项旨在测试重复鼻PD检测野生型CFTR功能的能力的独立系列实验中，在88%的对照（非CF）鼻PD中观察到Cl-分泌的证据，71%的PD超极化超过-5 mV。总之，这些结果表明，庆大霉素治疗可以抑制CF患者气道细胞中的提前终止突变，并在体内产生CFTR Cl-电导（通过鼻PD测量）的小幅增加。
Here we report the effects of gentamicin treatment on cystic fibrosis transmembrane regulator (CFTR) production and function in CF airway cells and patients with CF with premature stop mutations. Using immunocytochemical and functional [6-methoxy-N-(3-sulfopropyl) quinolinium (SPQ)-based] techniques, ex vivo exposure of airway cells from stop mutation CF patients led to the identification of surface-localized CFTR in a dose-dependent fashion. Next, five patients with CF with stop mutations and five CF control subjects were treated with parenteral gentamicin for 1 wk, and underwent repeated in vivo measures of CFTR function (nasal potential difference [PD] measurements and sweat chloride [Cl-] testing). During the treatment period, the number of nasal PD readings in the direction of Cl- secretion was increased approximately 3-fold in the stop mutation patient group compared with controls (p < 0.001), and four of five stop mutation patients with CF had at least one reading during gentamicin treatment with a Cl- secretory response of more than -5 mV (hyperpolarized). A response of this magnitude was not seen in any of the CF control subjects (p < 0.05). In an independent series of experiments designed to test the ability of repeat nasal PDs to detect wild-type CFTR function, evidence of Cl- secretion was seen in 88% of control (non-CF) nasal PDs, and 71% were more than -5 mV hyperpolarized. Together, these results suggest that gentamicin treatment can suppress premature stop mutations in airway cells from patients with CF, and produce small increases in CFTR Cl- conductance (as measured by the nasal PD) in vivo.