Pharmacological Characterization of Low Molecular Weight Biased Agonists at the Follicle Stimulating Hormone Receptor.
Pharmacological Characterization of Low Molecular Weight Biased Agonists at the Follicle Stimulating Hormone Receptor.
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促卵泡激素受体的低分子量偏性激动剂的药理学表征。
DOI：
10.3390/ijms22189850
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发表时间：
2021-09-12
影响因子：
5.6
通讯作者：
Reiter E
中科院分区：
文献类型：
--
作者：
De Pascali F;Ayoub MA;Benevelli R;Sposini S;Lehoux J;Gallay N;Raynaud P;Landomiel F;Jean-Alphonse F;Gauthier C;Pellissier LP;Crépieux P;Poupon A;Inoue A;Joubert N;Viaud-Massuard MC;Casarini L;Simoni M;Hanyaloglu AC;Nataraja SG;Yu HN;Palmer SS;Yvinec R;Reiter E
卵泡刺激素受体（FSHR）通过激活多种信号通路在生殖过程中发挥关键作用。低分子量（LMW）配体由偏向激动剂性质组成，是破译复杂信号传导机制的非常有价值的工具，因为它们允许选择性激活离散信号传导级联。然而，可用的LMW FSHR配体尚未完全表征。在这种情况下，我们探讨了药理多样性的三个苯甲酰胺和两个噻唑烷酮衍生物相比，FSH。在活细胞中使用BRET测定法生成Gαs、Gαq、Gαi、β-抑制蛋白2募集和cAMP产生的浓度/活性曲线。通过Western印迹分析ERK磷酸化，并使用荧光素酶报告基因测定评估CRE依赖性转录。所有测定均在野生型、Gαs或β-抑制蛋白1/2 CRISPR敲除HEK 293细胞中进行。通过使用操作模型计算每对读数的偏倚因子。我们的研究结果表明，与FSH相比，每种配体都表现出离散的药理学功效，范围从β-arrestin 2募集的超级激动剂到纯Gαs偏倚。有趣的是，LMW配体产生不同于FSH的动力学曲线（即，更快、更慢或瞬时，取决于配体），并与CRE依赖性转录相关。此外，在Gαs或β-arrestin基因缺失的细胞中观察到明显的系统偏倚。这种LMW特性是有用的药理学工具，可以更好地剖析FSHR激活的多种信号传导途径，并评估它们在细胞和生理病理水平上的相对贡献。
Follicle-stimulating hormone receptor (FSHR) plays a key role in reproduction through the activation of multiple signaling pathways. Low molecular weight (LMW) ligands composed of biased agonist properties are highly valuable tools to decipher complex signaling mechanisms as they allow selective activation of discrete signaling cascades. However, available LMW FSHR ligands have not been fully characterized yet. In this context, we explored the pharmacological diversity of three benzamide and two thiazolidinone derivatives compared to FSH. Concentration/activity curves were generated for Gαs, Gαq, Gαi, β-arrestin 2 recruitment, and cAMP production, using BRET assays in living cells. ERK phosphorylation was analyzed by Western blotting, and CRE-dependent transcription was assessed using a luciferase reporter assay. All assays were done in either wild-type, Gαs or β-arrestin 1/2 CRISPR knockout HEK293 cells. Bias factors were calculated for each pair of read-outs by using the operational model. Our results show that each ligand presented a discrete pharmacological efficacy compared to FSH, ranging from super-agonist for β-arrestin 2 recruitment to pure Gαs bias. Interestingly, LMW ligands generated kinetic profiles distinct from FSH (i.e., faster, slower or transient, depending on the ligand) and correlated with CRE-dependent transcription. In addition, clear system biases were observed in cells depleted of either Gαs or β-arrestin genes. Such LMW properties are useful pharmacological tools to better dissect the multiple signaling pathways activated by FSHR and assess their relative contributions at the cellular and physio-pathological levels.
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影响因子：
5.6
作者：
Nataraja S;Yu H;Guner J;Palmer S
通讯作者：
Palmer S
影响因子：
5.8
作者：
Danesi, R;LaRocca, RV;Myers, CE
通讯作者：
Myers, CE
影响因子：
3.6
作者：
Dias, James A.;Campo, Brice;Poli, Sonia M.
通讯作者：
Poli, Sonia M.
影响因子：
16.6
作者：
Klein Herenbrink C;Sykes DA;Donthamsetti P;Canals M;Coudrat T;Shonberg J;Scammells PJ;Capuano B;Sexton PM;Charlton SJ;Javitch JA;Christopoulos A;Lane JR
通讯作者：
Lane JR
影响因子：
4.8
作者：
Cottom, J;Salvador, LM;Hunzicker-Dunn, M
通讯作者：
Hunzicker-Dunn, M