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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要除了在细胞能量代谢中的作用之外，代谢物乳酸还影响多种生理过程并通过充当信号分子促进癌症进展。癌细胞经常经历代谢重编程，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 (HCAR1)，这是一种正在研究的 G 蛋白偶联受体通过驱动癌细胞增殖、刺激肿瘤血管化来促进肿瘤生长的受体抑制免疫监视。抑制 HCAR1 活性代表了一种有前途的癌症治疗策略，但缺乏靶向 HCAR1 的选择性药理学工具阻碍了我们靶向该受体的能力疾病并限制了我们对 HCAR1 生理学功能的理解。我们的长期目标是了解 HCAR1 配体识别的分子基础并利用该信息设计选择性小分子靶向该受体。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 HCAR1 配体促进纯化 HCAR1 和工程化 G 之间结合的能力蛋白（迷你 G 蛋白）或骆驼单链抗体片段（纳米抗体）。在目标 1 中，我们将检查目前可用的 HCAR1 激动剂和一组 mini-G 蛋白之间的变构耦合研究潜在 HCAR1 拮抗剂的作用机制。在目标 2 中，我们将识别构象识别 HCAR1 并确定其在 HCAR1 上的选择性和结合模式的选择性纳米抗体。在两者中目标，我们将确定形成稳定的 HCAR1 复合物的条件，用于未来 HCAR1 的结构研究多个州。这项工作将深入了解目前可用的 HCAR1 的分子药理学配体，发现新型纳米抗体来探测 HCAR1 功能，并实现未来基于结构的设计用于癌症治疗的 HCAR1 配体。