An Innovative High-Throughput Screening Approach for Discovery of Small Molecules That Inhibit TNF Receptors.
An Innovative High-Throughput Screening Approach for Discovery of Small Molecules That Inhibit TNF Receptors.
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DOI：
10.1177/2472555217706478
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发表时间：
2017-09
期刊：
影响因子：
--
通讯作者：
Sachs JN
中科院分区：
文献类型：
--
作者：
Lo CH;Vunnam N;Lewis AK;Chiu TL;Brummel BE;Schaaf TM;Grant BD;Bawaskar P;Thomas DD;Sachs JN
肿瘤坏死因子受体1（TNFR 1）是一种跨膜受体，其结合肿瘤坏死因子或光敏素-α，并且在调节炎症反应中起关键作用。这些配体的上调与炎性和自身免疫性疾病相关。目前的治疗通过螯合游离配体来减轻症状，但这可能会通过无意中抑制配体与脱靶受体的结合而引起不良副作用。因此，需要特异性靶向受体而不是配体的新的小分子。在这里，我们开发了一种在活细胞中表达的TNFR 1 FRET生物传感器，以筛选NIH临床收藏的化合物。我们使用了一个创新的高通量荧光寿命筛选平台，具有精致的空间和时间分辨率，以确定两个小分子化合物，扎鲁司特和三氯苯达唑，抑制TNFR 1诱导的IκBα降解和NF-κB激活。使用生物化学和计算对接方法来显示扎鲁司特破坏TNFR 1前配体组装结构域（PLAD）之间的相互作用，而三氯苯达唑起变构作用。重要的是，这两种化合物都不能抑制配体结合，这首次证明了通过靶向TNF受体-受体相互作用来抑制受体活化是可能的。这种策略应该普遍适用于TNFR超家族的其他成员，以及一般的寡聚受体。
Tumor necrosis factor receptor 1 (TNFR1) is a transmembrane receptor that binds tumor necrosis factor or lymphotoxin-alpha and plays a critical role in regulating the inflammatory response. Upregulation of these ligands is associated with inflammatory and autoimmune diseases. Current treatments reduce symptoms by sequestering free ligands, but this can cause adverse side effects by unintentionally inhibiting ligand binding to off-target receptors. Hence, there is a need for new small molecules that specifically target the receptors, rather than the ligands. Here, we developed a TNFR1 FRET biosensor expressed in living cells to screen compounds from the NIH Clinical Collection. We used an innovative high-throughput fluorescence lifetime screening platform that has exquisite spatial and temporal resolution to identify two small-molecule compounds, zafirlukast and triclabendazole, that inhibit the TNFR1-induced IκBα degradation and NF-κB activation. Biochemical and computational docking methods were used to show that zafirlukast disrupts the interactions between TNFR1 pre-ligand assembly domain (PLAD), whereas triclabendazole acts allosterically. Importantly, neither compound inhibits ligand binding, proving for the first time that it is possible to inhibit receptor activation by targeting TNF receptor-receptor interactions. This strategy should be generally applicable to other members of the TNFR superfamily, as well as to oligomeric receptors in general.
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影响因子：
168.9
作者：
Shakoor, N;Michalska, M;Block, JA
通讯作者：
Block, JA
影响因子：
--
作者：
Cornea RL;Gruber SJ;Lockamy EL;Muretta JM;Jin D;Chen J;Dahl R;Bartfai T;Zsebo KM;Gillispie GD;Thomas DD
通讯作者：
Thomas DD
影响因子：
--
作者：
Gruber SJ;Cornea RL;Li J;Peterson KC;Schaaf TM;Gillispie GD;Dahl R;Zsebo KM;Robia SL;Thomas DD
通讯作者：
Thomas DD
影响因子：
2.3
作者：
Fricke, Franziska;Malkusch, Sebastian;Heilemann, Mike
通讯作者：
Heilemann, Mike
影响因子：
1.6
作者：
Muretta, Joseph M.;Kyrychenko, Alexander;Thomas, David D.
通讯作者：
Thomas, David D.