Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice.
Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice.
复制标题
具有缩短的 R 结构域的囊性纤维化跨膜电导调节剂可挽救 CFTR-/- 小鼠的肠道表型。
DOI：
10.1073/pnas.1019752108
复制
发表时间：
2011
影响因子：
11.1
通讯作者：
Welsh,MichaelJ
中科院分区：
文献类型：
--
作者：
Ostedgaard,LyndaS;Meyerholz,DavidK;Vermeer,DanielW;Karp,PhilipH;Schneider,Lindsey;Sigmund,CurtD;Welsh,MichaelJ
基因转移为囊性纤维化的治疗提供了新的途径，腺相关病毒（AAV）是一种很有前途的载体。然而，AAV的包装能力限制了全长囊性纤维化跨膜传导调节因子（CFTR）cDNA连同其它调节和结构元件的包含。为了克服AAV大小的限制，我们最近开发了一种缺失R结构域N端部分（残基708-759，CFTRΔR）的缩短的CFTR，并发现它在体外保留了受调节的阴离子通道活性。为了验证CFTRΔR可以纠正体内缺陷的假设，我们产生了携带转基因的CFTR −/−小鼠，该转基因具有脂肪酸结合蛋白启动子，可驱动人CFTRΔR在肠道中的表达（CFTR−/−;TgΔR）。我们发现CFTR −/−;TgΔ R小鼠的肠隐窝表达CFTRΔR，并且肠在组织学上与WT小鼠相似。此外，与全长CFTR转基因一样，CFTR Δ R转基因产生CFTR Cl−电流并挽救CFTR −/−肠表型。这些结果表明，CFTR R结构域的N-末端部分对于体内肠生理学是不稳定的。因此，CFTRΔR可用于CF中的AAV介导的基因转移。
Gene transfer could provide a novel therapeutic approach for cystic fibrosis (CF), and adeno-associated virus (AAV) is a promising vector. However, the packaging capacity of AAV limits inclusion of the full-length cystic fibrosis transmembrane conductance regulator (CFTR) cDNA together with other regulatory and structural elements. To overcome AAV size constraints, we recently developed a shortened CFTR missing the N-terminal portion of the R domain (residues 708–759, CFTRΔR) and found that it retained regulated anion channel activity in vitro. To test the hypothesis that CFTRΔR could correct in vivo defects, we generatedCFTR−/−mice bearing a transgene with a fatty acid binding protein promoter driving expression of human CFTRΔR in the intestine (CFTR−/−;TgΔR). We found that intestinal crypts ofCFTR−/−;TgΔRmice expressed CFTRΔR and the intestine appeared histologically similar to that of WT mice. Moreover, like full-lengthCFTRtransgene, theCFTRΔRtransgene produced CFTR Cl−currents and rescued theCFTR−/−intestinal phenotype. These results indicate that the N-terminal part of the CFTR R domain is dispensable for in vivo intestinal physiology. Thus, CFTRΔR may have utility for AAV-mediated gene transfer in CF.