Højfeldt Jonas W, Cruz-Rodríguez Osvaldo, Imaeda Yasuhiro, Van Dyke Aaron R, Carolan James P, Mapp Anna K, Iñiguez-Lluhí Jorge A
Department of Chemistry (J.W.H.,Y.I., J.P.C., A.K.M.), University of Michigan, and Department of Pharmacology (O.C.-R., J.A.I.-L.), University of Michigan Medical School, Ann Arbor, Michigan 48109; and Department of Chemistry and Biochemistry (A.R.V.D.), Fairfield University, Fairfield, Connecticut 06824.
Mol Endocrinol. 2014 Feb;28(2):249-59. doi: 10.1210/me.2013-1343. Epub 2014 Jan 1.
Therapies based on conventional nuclear receptor ligands are extremely powerful, yet their broad and long-term use is often hindered by undesired side effects that are often part of the receptor's biological function. Selective control of nuclear receptors such as the glucocorticoid receptor (GR) using conventional ligands has proven particularly challenging. Because they act solely in an allosteric manner, conventional ligands are constrained to act via cofactors that can intrinsically partner with the receptor. Furthermore, effective means to rationally encode a bias for specific coregulators are generally lacking. Using the (GR) as a framework, we demonstrate here a versatile approach, based on bifunctional ligands, that extends the regulatory repertoire of GR in a deliberate and controlled manner. By linking the macrolide FK506 to a conventional agonist (dexamethasone) or antagonist (RU-486), we demonstrate that it is possible to bridge the intact receptor to either positively or negatively acting coregulatory proteins bearing an FK506 binding protein domain. Using this strategy, we show that extrinsic recruitment of a strong activation function can enhance the efficacy of the full agonist dexamethasone and reverse the antagonist character of RU-486 at an endogenous locus. Notably, the extrinsic recruitment of histone deacetylase-1 reduces the ability of GR to activate transcription from a canonical GR response element while preserving ligand-mediated repression of nuclear factor-κB. By providing novel ways for the receptor to engage specific coregulators, this unique ligand design approach has the potential to yield both novel tools for GR study and more selective therapeutics.
基于传统核受体配体的疗法极为强大,但其广泛且长期的使用常常受到不良副作用的阻碍,这些副作用往往是受体生物学功能的一部分。事实证明,使用传统配体对诸如糖皮质激素受体(GR)等核受体进行选择性调控极具挑战性。由于传统配体仅以别构方式起作用,它们只能通过可与受体内在结合的辅助因子发挥作用。此外,通常缺乏合理编码对特定共调节因子偏好的有效手段。在此,我们以GR为框架,展示了一种基于双功能配体的通用方法,该方法能以可控方式扩展GR的调控功能范围。通过将大环内酯类药物FK506与传统激动剂(地塞米松)或拮抗剂(RU - 486)相连,我们证明可以将完整的受体与带有FK506结合蛋白结构域的正向或负向作用的共调节蛋白相连接。运用这一策略,我们发现外源募集强大的激活功能可增强全激动剂地塞米松的效力,并在内源位点逆转RU - 486的拮抗剂特性。值得注意的是,外源募集组蛋白去乙酰化酶 - 1会降低GR从典型GR反应元件激活转录的能力,同时保留配体介导的对核因子 - κB的抑制作用。通过为受体提供与特定共调节因子结合的新方式,这种独特的配体设计方法有潜力为GR研究带来新工具,并开发出更具选择性的疗法。