In silico analysis and theratyping of an ultra-rare CFTR genotype (W57G/A234D) in primary human rectal and nasal epithelial cells.
In silico analysis and theratyping of an ultra-rare CFTR genotype (W57G/A234D) in primary human rectal and nasal epithelial cells.
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DOI：
10.1016/j.isci.2023.108180
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发表时间：
2023-11-17
期刊：
影响因子：
5.8
通讯作者：
Sorio, Claudio
中科院分区：
文献类型：
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作者：
Kleinfelder, Karina;Lotti, Virgini;Eramo, Adriana;Amato, Felice;Lo Cicero, Stefania;Castelli, Germana;Spadaro, Francesca;Farinazzo, Alessia;Dell'Orco, Daniele;Preato, Sara;Conti, Jessica;Rodella, Luca;Tomba, Francesco;Cerofolini, Angelo;Baldisseri, Elena;Bertini, Marina;Volpi, Sonia;Villella, Valeria Rachela;Esposito, Speranza;Zollo, Immacolata;Castaldo, Giuseppe;Laudanna, Carlo;Sorsher, Eric J.;Hong, Jeong;Joshi, Disha;Cutting, Garry;Lucarelli, Marco;Melotti, Paola;Sorio, Claudio
囊性纤维化（CF）的突变靶向治疗仍然不适合所有CF受试者，特别是对于携带CFTR基因型W57 G/A234 D（c.169T>G/c.701C>A）等罕见变异的病例。我们对这些变体对蛋白质稳定性的影响进行了计算机模拟分析，我们使用结肠样和重编程的鼻上皮细胞对其进行了功能表征。突变对囊性纤维化跨膜传导调节因子（CFTR）蛋白的影响通过蛋白质印迹、毛喉素诱导肿胀（FIS）和Ussing室分析进行分析。我们检测到在用CFTR调节剂VX 661 ± VX 445 ± VX 770处理后增加的残余CFTR功能，其在模型之间相关，并且与用CFTR校正剂处理后增加的CFTR蛋白水平相关。用VX 770进行的体内治疗将汗液氯化物浓度降低至非CF水平，增加了CFTR依赖性汗滴的数量，并在治疗27周后诱导预测的FEV1%绝对增加6%，表明CF中患者来源细胞的治疗相关性。非常罕见的CFTR变体/复杂基因型应作为1例中的N例进行研究患者源性鼻和结肠细胞中的治疗提供了重叠结果FIS和Ussing室试验提供了有关CFTR功能的补充信息A234 D变体负责对CFTR调节剂的反应治疗;人体标本;分子医学;细胞生物学的综合方面;药物信息学;计算机生物学
Mutation targeted therapy in cystic fibrosis (CF) is still not eligible for all CF subjects, especially for cases carrying rare variants such as the CFTR genotype W57G/A234D (c.169T>G/c.701C>A). We performed in silico analysis of the effects of these variants on protein stability, which we functionally characterized using colonoids and reprogrammed nasal epithelial cells. The effect of mutations on cystic fibrosis transmembrane conductance regulator (CFTR) protein was analyzed by western blotting, forskolin-induced swelling (FIS), and Ussing chamber analysis. We detected a residual CFTR function that increases following treatment with the CFTR modulators VX661±VX445±VX770, correlates among models, and is associated with increased CFTR protein levels following treatment with CFTR correctors. In vivo treatment with VX770 reduced sweat chloride concentration to non-CF levels, increased the number of CFTR-dependent sweat droplets, and induced a 6% absolute increase in predicted FEV1% after 27 weeks of treatment indicating the relevance of theratyping with patient-derived cells in CF. Very rare CFTR variants/complex genotypes should be studied as N-of-1 case Theratyping in patient-derived nasal and colon cells provides overlapping results FIS and Ussing chamber assays provide complementary information on CFTR function A234D variant is responsible for the response to CFTR modulators Therapy; Human specimen; Molecular medicine; Integrative aspects of cell biology; Pharmacoinformatics; In silico biology
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影响因子：
5
作者：
Matos AM;Jordan P;Matos P
通讯作者：
Matos P
影响因子：
10
作者：
Derichs, Nico;Sanz, Javier;Ballmann, Manfred
通讯作者：
Ballmann, Manfred
影响因子：
8.8
作者：
Berkers, Gitte;van Mourik, Peter;van der Ent, Cornelis K.
通讯作者：
van der Ent, Cornelis K.
影响因子：
8
作者：
Han, Sangwoo T.;Rab, Andras;Cutting, Garry R.
通讯作者：
Cutting, Garry R.
DOI：
10.1038/nrg3849
发表时间：
2015-01
期刊：
Nature reviews. Genetics
影响因子：
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作者：
通讯作者：
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