Nanobody- and mini-G protein-enabled molecular pharmacology of HCAR1

HCAR1 的纳米抗体和迷你 G 蛋白分子药理学

• 批准号: 10666999
• 负责人: William I Weis
• 金额: $ 15.46万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10666999
• 关键词: Acids; Address; Adipocytes; Affinity; Agonist; Antibodies; Automobile Driving; Binding; Biological Assay; Biosensor; Cell Proliferation; Cells; Central Nervous System; Complex; Coupling; Disease; Drug Design; Drug Screening; Energy Metabolism; Engineering; Foundations; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Immunoglobulin Fragments; Immunologic Surveillance; Interferometry; Kinetics; Libraries; Ligands; Lipid Bilayers; Lipids; Lipolysis; Malignant Neoplasms; Metabolic; Metabolism; Molecular; Molecular Conformation; Molecular Sieve Chromatography; Pharmacology; Physiological; Physiological Processes; Physiology; Production; Role; Signal Transduction; Signaling Molecule; Specificity; Stimulation of Cell Proliferation; Structure; Tumor Promotion; Vascularization; Waste Products; Work; Yeasts; angiogenesis; antagonist; cancer cell; cancer imaging; cancer therapy; cancer type; candidate identification; design; drug discovery; drug structure; extracellular; insight; nanobodies; nanodisk; novel; novel strategies; obesity treatment; pharmacologic; protein complex; receptor; reconstitution; small molecule; tool; tumor; tumor growth; tumor progression

Project Summary/Abstract Separate from its roles in cellular energy metabolism, the metabolite lactate influences multiple physiological processes and contributes to cancer progression by acting as a signaling molecule. Cancer cells often undergo metabolic reprogramming which results in increased lactate production and secretion. Extracellular lactate activates hydroxycarboxylic acid receptor 1 (HCAR1), an understudied G protein-coupled receptor that promotes tumor growth by driving cancer cell proliferation, stimulating tumor vascularization, and inhibiting immune surveillance. Inhibiting HCAR1 activity represents a promising strategy for cancer treatment, but the lack of selective pharmacological tools targeting HCAR1 hampers our ability to target this receptor in disease and limits our understanding of HCAR1 function in physiology. Our long-term goal is to understand the molecular basis for ligand recognition by HCAR1 and to use this information to design selective small molecules targeting this receptor. This project will lay the foundation for future structure-based drug discovery efforts by determining the ability of known HCAR1 ligands to promote binding between purified HCAR1 and engineered G proteins (mini-G proteins) or camelid single-chain antibody fragments (nanobodies). In Aim 1, we will examine the allosteric coupling between currently available HCAR1 agonists and a panel of mini-G proteins and investigate the mechanism of action of potential HCAR1 antagonists. In Aim 2, we will identify conformationally selective nanobodies that recognize HCAR1 and determine their selectivity and binding mode at HCAR1. In both aims, we will determine conditions to form stable HCAR1 complexes for future structural studies of HCAR1 in multiple states. This work will provide insight into the molecular pharmacology of currently available HCAR1 ligands, discover novel nanobodies to probe HCAR1 function, and enable future structure-based design of HCAR1 ligands for cancer treatment.

项目总结/摘要 除了其在细胞能量代谢中的作用，代谢物乳酸盐影响多种细胞能量代谢。 它通过充当信号分子参与生理过程并促进癌症进展。癌细胞 通常经历代谢重编程，这导致乳酸产生和分泌增加。 细胞外乳酸激活羟基羧酸受体1（HCAR 1），一种研究不足的G蛋白偶联的 通过驱动癌细胞增殖、刺激肿瘤血管形成和 抑制免疫监视。抑制HCAR 1活性代表了癌症治疗的有希望的策略， 但是缺乏针对HCAR 1的选择性药理学工具阻碍了我们靶向这种受体的能力， 疾病和限制了我们对HCAR 1在生理学中功能的理解。我们的长期目标是了解 HCAR 1识别配体的分子基础，并利用这些信息设计选择性小分子 针对这个受体。该项目将为未来基于结构的药物发现工作奠定基础， 确定已知HCAR 1配体促进纯化的HCAR 1和工程化的G 蛋白质（mini-G蛋白）或骆驼科动物单链抗体片段（纳米抗体）。在目标1中，我们将研究 目前可用的HCAR 1激动剂和一组mini-G蛋白之间的变构偶联， 研究潜在HCAR 1拮抗剂的作用机制。在目标2中，我们将确定构象 识别HCAR 1并确定其在HCAR 1处的选择性和结合模式的选择性纳米抗体。无论是 目的，我们将确定条件，以形成稳定的HCAR 1复合物，为未来的HCAR 1结构研究， 多个州。这项工作将提供深入了解目前可用的HCAR 1的分子药理学 配体，发现新的纳米抗体来探测HCAR 1功能，并使未来基于结构的设计成为可能。 用于癌症治疗的HCAR 1配体。