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可通过诱导和促进多巴胺神经传递 中唇多巴胺能途径，该途径修改了与奖励相关的行为，并且与 许多疾病的发展，包括精神分裂症，亨廷顿氏病，认知障碍和 帕金森氏病。越来越多的证据表明，这种重要的信号通路是建设性的 由多巴胺受体（DRS）调节。那使该膜蛋白家族的成员高度高 在临床前和临床研究的支持下，有希望的治疗靶标。 尽管人类DR（HDR）家族成员中的某些受体激动剂（例如多巴胺D1样 受体）正在深入研究治疗干预，其成功受到了极大的阻碍 由于依从性和效率差，或者由于相关的副作用。相关，多药理学 在最近的研究中发现了多巴胺D1样受体和其他HDR。基于 D1样受体（D1R和D5R）的结构不仅会揭示其信号传导机制，还将提供 可以探索的新理解以寄主有理药物设计。 HDR属于G蛋白偶联受体（GPCR）家族，该家族因产生困难而臭名昭著 衍射质量晶体对于通过X射线测定高分辨率结构至关重要 晶体学。该研发提案旨在为结构和 多巴胺D1样受体和HDR与G蛋白的复合物的功能研究，用于筛查高 靶向这些受体的亲和力纳米体，抗体和配体。该平台将涉及多个步骤 通过向前和向后的反馈系统紧密互连和循环。 PI现在有 还与其他研究小组建立了强有力的合作 方法，包括X射线无电子激光器，微晶电子衍射技术等。 提出了三个具体目的：（1）人类D1R（HD1R）在非活性状态下的结构/功能研究，（2） 使用X射线晶体学和计算生物学方法研究HD1R的假定活性状态， （3）建立优化的方法来确定与G蛋白的HD1R复合物的结构 使用EM成像的合作伙伴。这项研究的意义在多巴胺信号通路和 相关的药物发现研究：1）我们将深入了解多巴胺D1样受体功能和 变构调制，2）我们将能够广泛筛选以识别HDRS的新的高亲和力配体，3） 表征不同多巴胺接收器之间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）为在困难中应用冷冻技术铺平道路 膜蛋白将来的靶标。