Genotypic analysis of respiratory mucous sulfation defects in cystic fibrosis.
Genotypic analysis of respiratory mucous sulfation defects in cystic fibrosis.
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囊性纤维化呼吸道粘液硫酸盐化缺陷的基因型分析。
DOI：
10.1172/jci118372
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Engelhardt,JF
中科院分区：
文献类型：
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作者：
Zhang,Y;Doranz,B;Yankaskas,JR;Engelhardt,JF
囊性纤维化跨膜电导调节器(CFTR)的细胞内功能障碍已被提出改变内膜酸化。这一缺陷最广泛研究的后果是囊性纤维化(CF)呼吸道粘液糖蛋白的生化特性改变。然而，由于缺乏体内动物模型来验证这些假说，在没有慢性细菌感染的继发性影响的情况下，证实CF存在粘液加工缺陷的研究一直受到阻碍。人支气管异种移植模型在无继发性疾病影响(如杯状细胞增生)的情况下，可用于评估CF型和非CF型呼吸道上皮之间的病理生理学差异。在这项研究中，我们试图比较来自慢性支气管炎和非慢性支气管炎的人异种支气管移植物分泌的粘液糖蛋白的硫酸盐化程度。从13个独立的CF组织样本产生的异种移植的累积结果显示，与非CF配对的对照相比，有统计学意义的更高的硫酸盐化水平(1.7+/-0.18，P<；0.026)。这些发现增加了越来越多的知识，即CF呼吸道粘蛋白存在原发的硫酸盐化缺陷。基因型与粘液硫酸盐化程度的相关性显示，两种类型的CF组织的粘液硫化程度有统计学上的差异。这些研究的结果显示，与非CF型对照(P<；0.004，n=10)相比，来自五种明确的CF型的异种移植产生的粘液硫酸盐化水平高出2.0%/0.15%。有趣的是，与匹配的非CF对照组(n=3)相比，三个突变等位基因仍未确定的CF样本(Deltaoff8/未知或G551D/未知)在硫酸盐化水平上没有统计学差异。在26个最常见的CF突变的筛查中，这一未知的等位基因还没有被发现。这些结果为CFR群体内的等位基因变异提供了初步证据，这可能开始阐明CFTR与细胞内粘液加工缺陷有关的结构和功能。
Intracellular dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) has been proposed to alter endosomal acidification. The most widely studied consequence of this defect has been alterations in the biochemical properties of cystic fibrosis (CF) respiratory mucus glycoproteins. However, studies confirming the existence of mucous processing defects in CF have been hindered by the lack of in vivo animal models by which to test these hypotheses in the absence of secondary effects of chronic bacterial infection. The human bronchial xenograft model has been useful in evaluating the pathophysiologic differences between CF and non-CF airway epithelium, in the absence of secondary disease effects such as goblet cell hyperplasia. In this study we sought to compare the extent of sulfation within secreted mucus glycoproteins from CF and non-CF human bronchial xenografts. Cumulative results of xenografts generated from 13 independent CF tissue samples demonstrated a statistically significant higher level of sulfation (1.7 +/- 0.18, P < 0.026) as compared to non-CF paired controls. Such findings add to the growing body of knowledge that primary defects in sulfation exist in CF respiratory mucin. Correlation of genotype with the extent of mucus sulfation revealed two categories of CF tissues with statistically different mucus sulfation profiles. Results from these studies demonstrated a 2.0 +/- 0.15-fold higher level of mucus sulfation produced from xenografts of five defined CF genotypes as compared to non-CF controls (P < 0.004, n= 10). Interestingly, three CF samples for which one mutant allele remained undefined (deltaoff8/unknown or G551D/unknown) demonstrated no statistical difference in the level of sulfation as compared with matched non-CF controls (n= 3). This as yet unknown allele was not identified within a screen for the 26 most common CF mutations. These results provide preliminary evidence for allelic variation within the CF population which may begin to elucidate the structure-function of CFTR with regards to intracellular mucus processing defects.Images