1 Division of the National Toxicology Program, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, Durham, NC, USA.
2 National Institutes of Health Chemical Genomics Center, NIH Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA.
SLAS Discov. 2017 Jul;22(6):720-731. doi: 10.1177/2472555216689772. Epub 2017 Jan 31.
Estrogen-related receptor alpha (ERRα), the first orphan nuclear receptor discovered, is crucial for the control of cellular energy metabolism. ERRα and its coactivator, peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), are required for rapid energy production in response to environmental challenges. They have been implicated in the etiology of metabolic disorders such as type 2 diabetes and metabolic syndrome. ERRα also plays a role in the pathogenesis of breast cancer. Identification of compounds that modulate ERRα signaling may elucidate environmental factors associated with these diseases. Therefore, we developed stable cell lines containing an intact ERRα signaling pathway, with and without the coactivator PGC-1α, to use as high-throughput screening tools to detect ERRα modulators. The lentiviral PGC-1α expression constructs and ERRα multiple hormone response element (MHRE) reporters were introduced into HEK293T cells that express endogenous ERRα. A cell line expressing the reporter alone was designated "ERR." A second cell line expressing both reporter and PGC-1α was named "PGC/ERR." Initial screenings of the Library of Pharmacologically Active Compounds (LOPAC) identified 33 ERR and 22 PGC/ERR agonists, and 54 ERR and 15 PGC/ERR antagonists. Several potent ERRα agonists were dietary plant compounds (e.g., genistein). In conclusion, these cell lines are suitable for high-throughput screens to identify environmental chemicals affecting metabolic pathways and breast cancer progression.
雌激素相关受体 α（ERRα）是第一个被发现的孤儿核受体，对于控制细胞能量代谢至关重要。ERRα及其共激活因子过氧化物酶体增殖物激活受体 γ 共激活因子 1-α（PGC-1α）对于应对环境挑战时快速产生能量是必需的。它们与 2 型糖尿病和代谢综合征等代谢紊乱的病因有关。ERRα 在乳腺癌的发病机制中也发挥作用。鉴定调节 ERRα 信号的化合物可能阐明与这些疾病相关的环境因素。因此，我们开发了含有完整 ERRα 信号通路的稳定细胞系，包括有和没有共激活因子 PGC-1α 的细胞系，用作高通量筛选工具来检测 ERRα 调节剂。慢病毒 PGC-1α 表达构建体和 ERRα 多激素反应元件（MHRE）报告基因被引入表达内源性 ERRα 的 HEK293T 细胞。仅表达报告基因的细胞系被命名为“ERR”。表达报告基因和 PGC-1α 的第二个细胞系被命名为“PGC/ERR”。药理学活性化合物库（LOPAC）的初步筛选鉴定出 33 种 ERR 和 22 种 PGC/ERR 激动剂，以及 54 种 ERR 和 15 种 PGC/ERR 拮抗剂。几种有效的 ERRα 激动剂是膳食植物化合物（如染料木黄酮）。总之，这些细胞系适合用于高通量筛选，以鉴定影响代谢途径和乳腺癌进展的环境化学物质。
