A G-protein-coupled receptor-based sensor design platform for high throughput drug screening

基于 G 蛋白偶联受体的传感器设计平台，用于高通量药物筛选

基本信息

批准号：
10577416
负责人：
Kayla Elizabeth Kroning
金额：
$ 2.03万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10577416
关键词：
Address Adrenergic Receptor Agonist Binding Biological Assay Cell Culture Techniques Chemicals Collaborations Coupled Cyclic AMP Directed Molecular Evolution Dopamine D1 Receptor Drug Screening Drug Targeting Engineering Event FDA approved Family G-Protein-Coupled Receptors GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins Genomics Goals Human body Ligand Binding Ligands Mammalian Cell Measures Membrane Methods Michigan Modeling Molecular Conformation Monitor Noise Pathway interactions Pharmaceutical Preparations Pharmacologic Substance Physiological Processes Play Protein Engineering Radiolabeled Reagent Receptor Activation Research Role Signal Transduction Techniques Therapeutic Therapeutic Uses Universities Work antagonist beta-2 Adrenergic Receptors cost cost effective design drug use screening high throughput screening high-throughput drug screening improved kappa opioid receptors mu opioid receptors neuronal excitability novel novel therapeutics receptor recruit screening sensor therapeutic target

项目摘要

PROPOSAL SUMMARY G-protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors, comprising over 800 receptor types in the human body. GPCRs play crucial roles in various physiological processes. Consequently, GPCRs are the target of 34% of all FDA approved pharmaceuticals. Despite this significant number, more than half of GPCRs remain untargeted by FDA approved drugs. This is partially due to the lack of a cost-effective, high-throughput drug screening platform for monitoring the immediate signaling events following GPCR activation. To further explore the therapeutic potential of both the FDA drug-targeted and untargeted GPCRs, it is necessary to develop a new high-throughput GPCR drug screening platform with a high signal-to-noise. GPCR-based fluorescent sensors are advantageous for high-throughput drug screening because they do not require expensive reagents or substrates, enabling the screening of hundreds of compounds at low costs. I aim to engineer GPCR-based fluorescent sensors that can be used to screen drugs for both Gαi/o-coupled and Gαs- coupled GPCRs. To design and optimize this platform, I will engineer sensors for the kappa opioid receptor, mu opioid receptor, beta-2-adrenergic receptor, and the dopamine D1 receptor. I will use a combination of rational protein design and directed evolution to optimize these sensors. Then, I will perform a proof of principle drug screening with these sensors in collaboration with the Center of Chemical Genomics at the University of Michigan. I expect this platform to cost less and be easier to use than the current GPCR ligand drug screening platforms. The completion of this proposal will allow the engineering of a wide array of sensors for high throughput GPCR drug screening. This new fluorescent sensor design platform will enable the screening of novel drugs for GPCRs that can be used for therapeutic purposes.

提案摘要 G 蛋白偶联受体 (GPCR) 是最大的跨膜受体家族，包含 800 多种人体中的 GPCR 受体类型在各种生理过程中发挥着至关重要的作用。尽管这个数字很大，但 GPCR 是 34% 的 FDA 批准药物的目标。一半的 GPCR 仍未被 FDA 批准的药物靶向，部分原因是缺乏具有成本效益的药物。高通量药物筛选平台，用于监测 GPCR 后的即时信号事件为了进一步探索 FDA 药物靶向和非靶向 GPCR 的治疗潜力。有必要开发一种具有高信噪比的新型高通量GPCR药物筛选平台。基于 GPCR 的荧光传感器有利于高通量药物筛选，因为它们不需要我的目标是需要昂贵的试剂或底物，从而能够以低成本筛选数百种化合物。设计基于 GPCR 的荧光传感器，可用于筛选 Gαi/o 耦合和 Gαs- 药物为了设计和优化这个平台，我将为 kappa 阿片受体 mu 设计传感器。我将阿片受体、β2-肾上腺素受体和多巴胺D1受体合理组合使用。然后，我将进行原理药物的蛋白质设计和定向进化来优化这些传感器。与大学化学基因组学中心合作，使用这些传感器进行筛选我预计这个平台比目前的 GPCR 配体药物筛选成本更低且更易于使用。该提案的完成将允许设计各种用于高水平的传感器。这种新型荧光传感器设计平台将能够筛选新型药物。用于治疗目的的 GPCR。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A genetically encoded sensor with improved fluorescence intensity for opioid detection at cellular resolution.

一种基因编码传感器，具有改进的荧光强度，可在细胞分辨率下检测阿片类药物。

DOI：
发表时间：
2021-10-12
期刊：
影响因子：
0
作者：
Kroning, Kayla E;Li, Mingcheng;Petrescu, D Isabel;Wang, Wenjing
通讯作者：
Wang, Wenjing

A Modular Fluorescent Sensor Motif Used to Detect Opioids, Protein-Protein Interactions, and Protease Activity.

用于检测阿片类药物、蛋白质-蛋白质相互作用和蛋白酶活性的模块化荧光传感器基序。

DOI：
10.1021/acschembio.2c00364
发表时间：
2022-08-19
期刊：
ACS chemical biology
影响因子：
4
作者：
Kroning, Kayla E.;Li, Mingcheng;Shen, Jiaqi;Fiel, Hailey;Nassar, Manon;Wang, Wenjing
通讯作者：
Wang, Wenjing