P2X4受体参与酒精使用障碍的结构基础及新型分子发现

酒精使用障碍（alcohol use disorders, AUDs）因千百年来多国流行的饮酒文化已逐渐成为全球人群中高发的精神疾病之一。目前用于治疗AUDs的药物少且有效性低，因此迫切需要开发新的高效药物。越来越多的证据表明P2X4受体在酒精使用障碍中扮演重要角色，然而乙醇抑制P2X4的分子基础仍未明确。本项目前期的预实验结果揭示乙醇抑制P2X4完全依赖于生理浓度下的胞外钙离子引起的处于变构抑制状态的P2X4，并发现和验证了钙调节乙醇结合的关键位点，进而拟综合运用分子生化、膜生物物理等手段，研究P2X4、钙、乙醇三者相互作用的机制，从而揭示P2X4参与AUDs的结构基础；通过已验证的钙调节乙醇关键位点的基因敲入小鼠及行为学实验，进一步验证P2X4参与AUDs的分子基础；并拟通过寻找新的化合物用于移除钙离子的作用从而阻断酒精对P2X4的抑制作用，以期为AUDs提供新的治疗策略。

Due to the drinking cultures in many countries, alcohol use disorders (AUDs) become one of the most common mental illnesses in the global population. The scarcity of current drugs therapy for AUDs and their limited efficacy have made it urgent to seek new effective drugs. Growing evidence supports that purinergic P2X4 receptors play important roles in AUDs, however, the structural basis for the interaction between the alcohol and P2X4 remains unclear. Our preliminary studies demonstrated that P2X4 can be only modulated by the ethanol at the allosteric inhibition state caused by extracellular Ca2+. And the key site involved in extracellular Ca2+-mediated regulation on the ethanol’s binding was also identified. Therefore, by a combination of using molecular, biochemical and biophysical approaches, our project is aiming to uncover the mechanism underlying the crosstalk between the ethanol and calcium in P2X4 receptors, which will be helpful to elucidate the role of P2X4 in AUDs. Further combining the behavioral analysis of knockin mice with a mutation in the Ca2+-binding site, we intend to further verify the molecular basis of P2X4 receptors in the AUDs. Based on this information, we further intend to discover and develop novel small molecules to interrupt the interactions among the alcohol, Ca2+ and P2X4 receptors. This project is expected to provide new treatment strategies for AUDs.