Structural Biology of Dopamine Signaling

多巴胺信号传导的结构生物学

• 批准号: 10543124
• 负责人: Wei Liu
• 金额: $ 31.98万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10543124
• 关键词: Address; Adherence; Affinity; Agonist; Antibodies; Area; Arizona; Attentional deficit; Behavior; Binding; Clinical Research; Cognition Disorders; Collaborations; Complex; Computational Biology; Cryoelectron Microscopy; Crystallization; Development; Disease; Docking; Dopamine; Dopamine D1 Receptor; Dopamine Receptor; Drug Design; Family; Family member; Feedback; Foundations; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Human; Huntington Disease; Hyperactivity; Image; Laboratories; Ligand Binding; Ligands; Membrane Proteins; Mental Depression; Molecular; Nerve; Nervous System; Neurons; Neurotransmitters; Parkinson Disease; Pathway interactions; Play; Process; Protein Family; Proteins; Receptor Signaling; Research; Resolution; Rewards; Roentgen Rays; Role; Schizophrenia; Signal Pathway; Signal Transduction; Signaling Protein; Site-Directed Mutagenesis; Solid; Structure; Structure-Activity Relationship; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Therapeutic Studies; Universities; Work; X-Ray Crystallography; addiction; antagonist; computer studies; design; dopamine D3 receptor; drug discovery; electron diffraction; insight; knowledge base; medical schools; member; molecular modeling; nanobodies; nervous system disorder; neurotransmission; novel therapeutics; positive allosteric modulator; preclinical study; protein complex; protein structure; rational design; receptor; receptor binding; research and development; screening; side effect; structural biology; success; therapeutic target; three dimensional structure; x-ray free-electron laser

Abstract Human dopamine signaling pathway induces and facilitates dopamine neurotransmission through the mesolimbic dopaminergic pathway, which modifies reward-related behaviors and is associated with the development of many diseases, including schizophrenia, Huntington's disease, cognitive disorders and Parkinson’s disease. Mounting evidence suggests that this important signaling pathway is constructively regulated by the dopamine receptors (DRs). Thus, making members of this membrane protein family highly promising therapeutic targets as supported by both pre-clinical and clinical studies. Although some agonists of receptors in the human DR (hDR) family members (e.g. for dopamine D1-like receptors) are being intensively studied for therapeutic intervention, their success has been greatly hampered due to poor adherence and efficacy, or due to associated side effects. Relatedly, the polypharmacology of dopamine D1-like receptor and other hDRs have been discovered in recent studies. New knowledge based on structures of D1-like receptors (D1R and D5R) will not only reveal their signaling mechanisms, but also provide new understanding that can be exploited to facilitate rational drug design. hDRs belong to the G protein-coupled receptor (GPCR) family which is notorious for difficulties in generating diffraction-quality crystals that are essential for the determination of high-resolution structures by X-ray crystallography. This R&D proposal aims to develop a comprehensive and robust platform for structural and functional studies of dopamine D1-like receptors and complexes of hDRs with G proteins, for screening high- affinity nanobodies, antibodies, and ligands targeting these receptors. This platform will involve multiple steps that are closely interconnected and looped through a forward and backward feedback system. The PI has now also established strong collaborations with other research groups with different expertise as alternative approaches, including X-ray free electron laser, Microcrystal Electron Diffraction technique, etc. Three specific aims are proposed: (1) Structure/function studies of human D1R (hD1R) in the inactive state, (2) Using X-ray crystallography and computational biology approaches to study the putative active state of hD1R, and (3) Establishing optimized approaches for determining the structure of complexes of hD1R with G protein partners using EM imaging. The significance of this study is multi-fold on dopamine signaling pathway and related drug discovery studies: 1) we will gain insights into dopamine D1-like receptor functionalities and allosteric modulations, 2) we will be able to screen extensively to identify new high-affinity ligands for hDRs, 3) characterize the mechanisms of DR signaling and ligand selection between different dopamine receptor subfamilies, 4) stimulate hDR structure-based drug design, 5) examine hDRs/G protein complex signaling and reveal the activation mechanism, and 6) pave the road for the application of cryoEM technology on difficult membrane protein targets in the future.

抽象的 人类多巴胺信号通路通过以下途径诱导和促进多巴胺神经传递 中脑边缘多巴胺能通路，它改变奖励相关的行为并与 许多疾病的发展，包括精神分裂症、亨廷顿舞蹈病、认知障碍和 越来越多的证据表明这一重要的信号通路具有建设性。 因此，该膜蛋白家族的成员受到多巴胺受体（DR）的调节。 有前景的临床前和临床研究支持的治疗目标。 尽管人类 DR (hDR) 家族成员中的一些受体激动剂（例如多巴胺 D1 样受体） 受体）正在被深入研究用于治疗干预，但它们的成功受到了极大的阻碍 由于依从性和疗效不佳，或由于相关的副作用。 最近的研究中发现了多巴胺D1样受体和其他hDRs的新知识。 D1 样受体（D1R 和 D5R）的结构不仅将揭示其信号传导机制，而且还提供 新的认识可用于促进合理的药物设计。 hDR 属于 G 蛋白偶联受体 (GPCR) 家族，该家族因难以生成而臭名昭著。 衍射质量晶体对于 X 射线确定高分辨率结构至关重要 该研发计划旨在开发一个全面且强大的结构和晶体学平台。 多巴胺 D1 样受体和 hDR 与 G 蛋白复合物的功能研究，用于筛选高 该平台将涉及多个步骤。 它们通过前向和后向反馈系统紧密相连和循环。 还与具有不同专业知识的其他研究小组建立了强有力的合作作为替代方案 方法，包括X射线自由电子激光、微晶电子衍射技术等。 提出了三个具体目标：（1）非活性状态下人类 D1R（hD1R）的结构/功能研究，（2） 使用 X 射线晶体学和计算生物学方法来研究 hD1R 的假定活性状态， (3) 建立确定 hD1R 与 G 蛋白复合物结构的优化方法 这项研究对于多巴胺信号通路的意义是多方面的。 相关药物发现研究：1）我们将深入了解多巴胺 D1 样受体功能和 变构调节，2) 我们将能够广泛筛选以确定新的 hDR 高亲和力配体，3) 表征不同多巴胺受体之间 DR 信号传导和配体选择的机制 亚家族，4) 刺激基于 hDR 结构的药物设计，5) 检查 hDRs/G 蛋白复合物信号传导和 揭示激活机制，6）为冷冻电镜技术在疑难问题上的应用铺平道路 膜蛋白是未来的靶标。