Nonselective TRPC channel inhibition and suppression of aminoglycoside-induced premature termination codon readthrough by the small molecule AC1903.
Nonselective TRPC channel inhibition and suppression of aminoglycoside-induced premature termination codon readthrough by the small molecule AC1903.
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DOI：
10.1016/j.jbc.2021.101546
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Bon RS
中科院分区：
文献类型：
--
作者：
Baradaran-Heravi A;Bauer CC;Pickles IB;Hosseini-Farahabadi S;Balgi AD;Choi K;Linley DM;Beech DJ;Roberge M;Bon RS
无义突变发生在约11%的遗传性疾病患者中，其引入导致截短蛋白质并促进无义介导的mRNA衰变的提前终止密码子（PTC）。氨基糖苷类如G418允许PTC通读，因此可用于解决该问题。然而，它们的效果在患者之间是可变的，使得氨基糖苷类的临床使用具有挑战性。在这项研究中，我们测试了TRPC非选择性阳离子通道是否有助于通过控制细胞摄取氨基糖苷类药物的可变PTC通读效应。事实上，最近报道的选择性TRPC 5抑制剂AC 1903在DMS-114癌细胞系和大疱性表皮交界（JEB）患者衍生的角质形成细胞中始终抑制G418摄取和G418诱导的PTC通读。有趣的是，AC 1903在DMS-114细胞中的作用被非选择性TRPC抑制剂模拟，但没有被TRPC 145（Pico 145，GFB-8438）或TRPC 3/6/7（SAR 7334）的充分表征的抑制剂模拟，这表明AC 1903可能通过其他或未定义的靶点起作用。事实上，在我们的实验中，AC 1903抑制多种TRPC通道，包括TRPC 3、TRPC 4、TRPC 5、TRPC 6、TRPC 4-C1和TRPC 5-C1，以及A498肾癌细胞中的内源性TRPC 1：C4通道，所有通道的微摩尔IC 50值均较低（1.8-18 μM）。我们还表明，AC 1903抑制TRPV 4通道，但对TRPV 1有微弱或无影响，对非选择性阳离子通道PIEZO 1无影响。我们的研究表明，AC 1903具有以前未识别的靶点，在解释该化合物的实验结果时需要考虑这些靶点。此外，我们的数据加强了非选择性钙通道参与氨基糖苷类摄取的假设。
Nonsense mutations, which occur in ∼11% of patients with genetic disorders, introduce premature termination codons (PTCs) that lead to truncated proteins and promote nonsense-mediated mRNA decay. Aminoglycosides such as G418 permit PTC readthrough and so may be used to address this problem. However, their effects are variable between patients, making clinical use of aminoglycosides challenging. In this study, we tested whether TRPC nonselective cation channels contribute to the variable PTC readthrough effect of aminoglycosides by controlling their cellular uptake. Indeed, a recently reported selective TRPC5 inhibitor, AC1903, consistently suppressed G418 uptake and G418-induced PTC readthrough in the DMS-114 cancer cell line and junctional epidermolysis bullosa (JEB) patient-derived keratinocytes. Interestingly, the effect of AC1903 in DMS-114 cells was mimicked by nonselective TRPC inhibitors, but not by well-characterized inhibitors of TRPC1/4/5 (Pico145, GFB-8438) or TRPC3/6/7 (SAR7334), suggesting that AC1903 may work through additional or undefined targets. Indeed, in our experiments, AC1903 inhibited multiple TRPC channels including TRPC3, TRPC4, TRPC5, TRPC6, TRPC4–C1, and TRPC5–C1, as well as endogenous TRPC1:C4 channels in A498 renal cancer cells, all with low micromolar IC50 values (1.8–18 μM). We also show that AC1903 inhibited TRPV4 channels, but had weak or no effects on TRPV1 and no effect on the nonselective cation channel PIEZO1. Our study reveals that AC1903 has previously unrecognized targets, which need to be considered when interpreting results from experiments with this compound. In addition, our data strengthen the hypothesis that nonselective calcium channels are involved in aminoglycoside uptake.
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DOI：
10.1002/anie.201411511
发表时间：
2015-03-16
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Akbulut Y;Gaunt HJ;Muraki K;Ludlow MJ;Amer MS;Bruns A;Vasudev NS;Radtke L;Willot M;Hahn S;Seitz T;Ziegler S;Christmann M;Beech DJ;Waldmann H
通讯作者：
Waldmann H
影响因子：
4.1
作者：
通讯作者：
--
DOI：
10.1164/ajrccm.163.7.2004001
发表时间：
2001-06-01
影响因子：
24.7
作者：
Clancy, JP;Bobök, Z;Bedwell, DM
通讯作者：
Bedwell, DM
影响因子：
14.9
作者：
BURKE, JF;MOGG, AE
通讯作者：
MOGG, AE
影响因子：
14.9
作者：
Baradaran-Heravi A;Balgi AD;Hosseini-Farahabadi S;Choi K;Has C;Roberge M
通讯作者：
Roberge M