Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), Università degli Studi di Palermo, Viale delle Scienze Ed. 16-17, 90128 Palermo, Italy.
Centro di OncoBiologia Sperimentale (COBS), via San Lorenzo Colli, 90145 Palermo, Italy.
Int J Mol Sci. 2020 Sep 3;21(17):6420. doi: 10.3390/ijms21176420.
Cystic fibrosis (CF) patients develop a severe form of the disease when the cystic fibrosis transmembrane conductance regulator (CFTR) gene is affected by nonsense mutations. Nonsense mutations are responsible for the presence of a premature termination codon (PTC) in the mRNA, creating a lack of functional protein. In this context, translational readthrough-inducing drugs (TRIDs) represent a promising approach to correct the basic defect caused by PTCs. By using computational optimization and biological screening, we identified three new small molecules showing high readthrough activity. The activity of these compounds has been verified by evaluating CFTR expression and functionality after treatment with the selected molecules in cells expressing nonsense-CFTR-mRNA. Additionally, the channel functionality was measured by the halide sensitive yellow fluorescent protein (YFP) quenching assay. All three of the new TRIDs displayed high readthrough activity and low toxicity and can be considered for further evaluation as a therapeutic approach toward the second major cause of CF.
囊性纤维化（CF）患者在囊性纤维化跨膜电导调节因子（CFTR）基因受到无义突变影响时会发展为严重形式的疾病。无义突变导致 mRNA 中出现提前终止密码子（PTC），从而导致缺乏功能性蛋白质。在这种情况下，翻译通读诱导药物（TRIDs）代表了一种纠正由 PTC 引起的基本缺陷的有前途的方法。通过使用计算优化和生物筛选，我们鉴定出三种具有高通读活性的新型小分子。通过在用表达无义-CFTR-mRNA 的细胞中用选定的分子处理后评估 CFTR 表达和功能，验证了这些化合物的活性。此外，通过卤化物敏感的黄色荧光蛋白（YFP）猝灭测定测量通道功能。这三种新型 TRIDs 均表现出高通读活性和低毒性，可考虑进一步评估作为 CF 的第二大主要病因的治疗方法。
