Biosensor for Screening specific Interactions with AhR

了解化合物和药物与芳烃受体（AhR）和雌激素受体（ER）信号通路相互作用的机制对于识别潜在的环境和内分泌干扰物以及筛选用于治疗的化合物都很重要。AhR和ER之间的串扰的存在导致某些AhR配体在不与ER结合的情况下发挥雌激素性内分泌干扰活性。 研究这些机制的现有方法繁琐且耗时，并且对于基于细胞的方法，通过细胞代谢的化合物降解显著影响结果。 最近，IA已经开发了生物传感器，用于研究在激动剂和拮抗剂存在下ER-α和-β与雌激素核反应元件（ERE）之间的实时缔合和解离动力学。 IA建议扩展这项技术，以开发化学和方法，使生物传感器能够应用于研究复杂的AhR介导的信号通路。 在第一阶段，将开发用于评估AhR与配体、辅助蛋白和二恶英反应元件（DRE）相互作用的生物传感器。 在第二阶段，生物传感器将用于分析ER和AhR之间的串扰，并创建基于hER和AhR机制预测化合物效果的筛选方法。 拟议的商业应用： 生物传感器，允许研究化合物（激动剂，拮抗剂，辅助蛋白）的AhR的实时结合动力学将为研究人员提供强大的工具，筛选环境内分泌干扰物和设计，开发和测试新的药物治疗癌症。

Understanding the mechanisms by which chemical compounds and drugs interact with both the aryl hydrocarbon receptor (AhR) and estrogen receptor (ER) signaling pathways is important both for identifying potential environmental and endocrine disruptors and for screening compounds for therapeutic use. The existence of cross-talk between AhR and ER causes certain AhR ligands to exert estrogenic endocrine disrupting activity in the absence of binding to ER. Existing methods for studying these mechanisms are cumbersome and time consuming, and for cell-based methods, compound degradation by cellular metabolism significantly affects results. Recently, IA has developed biosensors for studying real-time association and dissociation kinetics between ER-alpha & -beta, and estrogenic nuclear response element (ERE) in the presence of agonists and antagonists. IA proposes to extend this technology to develop chemistries and methods which will allow the biosensors to be applied to study the complex AhR-mediated signaling pathways. During Phase I, biosensors for assessing the interaction of AhR with ligands, accessory proteins, and dioxin response element (DRE) will be developed. During Phase II, biosensors will be used to analyze cross-talk between ER and AhR, and to create screening methods for predicting effects of compounds based on hER & AhR mechanisms. PROPOSED COMMERCIAL APPLICATIONS: Biosensors which allow studying the real-time binding kinetics of compounds (agonists, antagonists, accessory proteins) to the AhR will provide researchers with powerful tools for screening environmental endocrine disruptors and for designing, developing, and testing new pharmaceuticals for treatment of cancer.