Functional Genomics of G Protein-Coupled Receptors

G 蛋白偶联受体的功能基因组学

• 批准号: 10010909
• 负责人: Sriram Kosuri
• 金额: $ 24.69万
• 依托单位: OCTANT, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2020-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10010909
• 关键词: Affect; Agonist; Amino Acids; Amplifiers; Bar Codes; Binding; Biological Assay; Blindness; Calcium; Calcium Signaling; Calcium-Sensing Receptors; Cell Line; Cell physiology; Cells; Chemicals; Complex; Cyclic AMP; Cyclic AMP Receptors; DNA biosynthesis; DNA sequencing; Disease; Drug Targeting; Engineering; Environment; Epitopes; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genetic; Genetic Transcription; Genetic Variation; Genome engineering; Health; Hormones; Human; Human Cell Line; Individual; Informatics; Libraries; Mammalian Cell; Measures; Mediator of activation protein; Melanocortin 4 Receptor; Membrane; Methodology; Methods; Missense Mutation; Molecular; Mutation; Mutation Analysis; Natural Product Drug; Nonsense Mutation; Obesity; Odors; Oligonucleotides; Output; Pathway interactions; Pharmaceutical Preparations; Pharmacogenomics; Phase; Physiological; Physiology; Play; Population; Proteins; Proxy; Receptor Signaling; Reporter; Role; Running; Signal Pathway; Signal Transduction; Stimulus; System; Technology; Therapeutic Intervention; United States Food and Drug Administration; Variant; alpha-Melanocyte stimulating hormone; base; burden of illness; design; functional genomics; gain of function; gene function; genetic variant; genome editing; improved; innovation; insight; loss of function; mutant; next generation; next generation sequencing; novel; novel therapeutics; protective effect; receptor; receptor expression; receptor function; response; targeted treatment; technology development

G protein-coupled receptors (GPCRs) are central mediators of mammalian cells’ ability to sense and respond to their environment. The 813 human GPCRs are the largest class of membrane receptors, are central mediators of cell physiology, and are the target of ~34% of all U.S. Food and Drug Administration (FDA)-approved drugs and ~60% of prescriptions. Here we will leverage recent advances in DNA synthesis, genome editing, next-generation DNA sequencing, and informatics develop a platform to build thousands of individual mutations to GPCRs and experimentally characterize their effects in a novel assay that can be done in a simple pooled format in human cell lines. We will develop methodologies to characterize how these mutational libraries functionally signal through the two major pathways that GPCRs signal through, the cyclic AMP and calcium signaling pathways. The profiles will give us the ability to better understand the functional consequences of genetic variation in GPCRs and how they might be impacting diseases and drug responses. In addition, analysis of these mutational profiles will give us insights on how to target these receptors for making new drugs. The technologies developed here should be broadly and directly applicable to the vast majority of GPCRs, as well as other important classes of drug targets where function can be assessed by transcriptional reporters in human cell lines.

G 蛋白偶联受体 (GPCR) 是哺乳动物细胞感知和响应能力的核心介质 他们的环境。 813 人类 GPCR 是最大的一类膜受体，是中枢性的 细胞生理学的介质，是美国食品和药物管理局约 34% 的目标 (FDA) 批准的药物和约 60% 的处方。在这里我们将利用 DNA 合成的最新进展， 基因组编辑、下一代 DNA 测序和信息学开发了一个平台来构建数千个 GPCR 的个体突变并通过一种新的检测方法通过实验表征其影响 在人类细胞系中以简单的汇集形式。我们将开发方法来描述这些 突变文库通过 GPCR 信号传导的两条主要途径（循环）发挥功能性信号传导 AMP 和钙信号通路。这些配置文件将使我们能够更好地理解功能 GPCR 遗传变异的后果以及它们如何影响疾病和药物反应。 此外，对这些突变谱的分析将使我们了解如何靶向这些受体 制造新药。这里开发的技术应该广泛、直接适用于广大的领域。 大多数 GPCR 以及其他重要类别的药物靶标，其功能可以通过以下方式评估 人类细胞系中的转录报告基因。