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.

项目摘要/摘要与其在细胞能量代谢中的作用分开，代谢物乳酸会影响多种生理过程，并通过充当信号分子而导致癌症进展。癌细胞通常会经过代谢重编程，从而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），一种研究的G蛋白偶联通过驱动癌细胞增殖，刺激肿瘤血管形成和促进肿瘤生长的受体抑制免疫监测。抑制HCAR1活性代表了癌症治疗的有前途的策略，但是缺乏针对HCAR1的选择性药理学工具阻碍了我们在疾病和限制了我们对HCAR1生理功能的理解。我们的长期目标是了解 HCAR1识别配体识别的分子基础，并使用此信息设计选择性小分子靶向该受体。该项目将为以后的基于结构的药物发现努力奠定基础确定已知的HCAR1配体促进纯化HCAR1和工程G之间结合的能力蛋白质（迷你G蛋白）或骆驼单链抗体片段（纳米化）。在AIM 1中，我们将检查当前可用的HCAR1激动剂与一系列迷你G蛋白和一系列的变构耦合研究潜在的HCAR1拮抗剂的作用机理。在AIM 2中，我们将在构象上确定识别HCAR1并在HCAR1处确定其选择性和结合模式的选择性纳米结论。在这两个中目的，我们将确定形成稳定的HCAR1复合物的条件多个状态。这项工作将洞悉目前可用的HCAR1的分子药理学配体，发现新颖的纳米构造以探测HCAR1功能，并启用以后的基于结构的设计 HCAR1配体用于癌症治疗。