Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, CEINGE - Biotecnologie Avanzate, Naples, Italy.
Cell Biology and Disease Mechanisms Program, Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Italy.
Hum Mutat. 2019 Jun;40(6):742-748. doi: 10.1002/humu.23741. Epub 2019 Mar 28.
Pharmacological rescue of mutant cystic fibrosis transmembrane conductance regulator (CFTR) in cystic fibrosis (CF) depends on the specific defect caused by different mutation classes. We asked whether a patient with the rare p.Gly970Asp (c.2909G>A) mutation could benefit from CFTR pharmacotherapy since a similar missense mutant p.Gly970Arg (c.2908G>C) was previously found to be sensitive to potentiators in vitro but not in vivo. By complementary DNA transfection, we found that both mutations are associated with defective CFTR function amenable to pharmacological treatment. However, analysis of messenger RNA (mRNA) from patient's cells revealed that c.2908G>C impairs RNA splicing whereas c.2909G>A does not perturb splicing and leads to the expected p.Gly970Asp mutation. In agreement with these results, nasal epithelial cells from the p.Gly970Asp patient showed significant improvement of CFTR function upon pharmacological treatment. Our results underline the importance of controlling the effect of CF mutation at the mRNA level to determine if the pharmacotherapy of CFTR basic defect is appropriate.
在囊性纤维化（CF）中，突变型囊性纤维化跨膜电导调节因子（CFTR）的药物恢复依赖于不同突变类引起的特定缺陷。我们询问了一位罕见的 p.Gly970Asp（c.2909G>A）突变患者是否可以从 CFTR 药物治疗中获益，因为先前发现类似的错义突变 p.Gly970Arg（c.2908G>C）在体外对增敏剂敏感，但在体内不敏感。通过 cDNA 转染，我们发现这两种突变都与可通过药物治疗纠正的 CFTR 功能缺陷有关。然而，对患者细胞的信使 RNA（mRNA）的分析表明，c.2908G>C 损害 RNA 剪接，而 c.2909G>A 不会干扰剪接，导致预期的 p.Gly970Asp 突变。与这些结果一致，p.Gly970Asp 患者的鼻上皮细胞在药物治疗后 CFTR 功能显著改善。我们的结果强调了在 mRNA 水平控制 CF 突变的影响以确定 CFTR 基本缺陷的药物治疗是否合适的重要性。
