Spatiotemporal signaling and trafficking of the mu-opioid receptor

mu-阿片受体的时空信号传导和运输

基本信息

批准号：
10895814
负责人：
Ruth Huttenhain
金额：
$ 60.41万
依托单位：
STANFORD UNIVERSITY
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：
2022-07-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10895814
关键词：
Accidents Address Adverse effects Agonist Analgesics Biochemical Biological Assay Biology CRISPR interference CRISPR screen Cell Line Cell membrane Cell model Cells Cessation of life Chemicals Clinical Clustered Regularly Interspaced Short Palindromic Repeats Complex Cyclic AMP Data Down-Regulation Endosomes Epidemic GTP-Binding Proteins Gene Expression Regulation Genes Genetic Screening Genetic Transcription Genomics Goals Golgi Apparatus Human Induced pluripotent stem cell derived neurons Knock-out Knowledge Label Ligand Binding Ligands Location MAPK Signaling Pathway Pathway Maps Mass Spectrum Analysis Mediating Membrane Modality Molecular Neurons Opiate Addiction Opioid Opioid Receptor Opioid agonist Overdose Pain Pathway interactions Peptides Pharmaceutical Preparations Phenotype Phosphorylation Proteins Proteome Proteomics Receptor Activation Receptor Signaling Recycling Reporter Role Signal Pathway Signal Transduction Synaptic Vesicles Testing Transducers Validation Ventilatory Depression Work abuse liability addiction antagonist beta-arrestin data integration design experimental study functional genomics gene discovery insight knock-down mu opioid receptors new therapeutic target novel novel therapeutic intervention opioid abuse opioid epidemic opioid use phosphoproteomics presynaptic protein function receptor receptor internalization response side effect spatiotemporal synaptic inhibition trafficking

项目摘要

SUMMARY Opioids are the most effective analgesics but are associated with severe side effects including respiratory depression, tolerance, and addiction. These factors helped cause the opioid abuse epidemic in the US, making drug overdose the leading cause of accidental death in the US. Thus, the identification of safer analgesics with diminished side effects and abuse potential is critical to address the ongoing crisis. Clinically used opioids predominantly exert both their analgesic and adverse effects through their action on the µ-opioid receptor (MOR). While several approaches were taken towards safer analgesics, these efforts are limited by a lack of understanding the complex biochemical networks engaged and activated by MOR in response to ligand binding. This proposal builds on recent evidence suggesting that (1) MOR signaling is dependent on the interplay between subcellular localization and membrane trafficking in a ligand-specific manner and (2) MOR shows ligand- dependent effects on its protein interaction network and the signaling pathways it activates. Thus, delineating the MOR-initiated signaling pathways for endogenous peptides and addictive opioids and how these are coordinated by receptor location and trafficking provides potential new strategies for therapeutic modalities and safer analgesics. The overarching goal of this proposal is to combine quantitative proteomics, functional genomics, and opioid receptor biology to systematically discover and characterize regulators of MOR signaling and trafficking in human induced pluripotent stem cell-derived neurons. We will combine proximity labeling mass spectrometry and quantitative phosphoproteomics to systematically delineate interaction networks that MOR engages and map the signaling pathways it activates. To study the functional role of proteomic targets in MOR signaling and trafficking, we will develop and apply reporter assays for receptor signaling and trafficking in CRISPRi gene regulation screens. Finally, we will test mechanistic hypotheses from proteomic and genetic screens on how novel regulators of trafficking and signaling fine tune the cellular response of MOR activation. Our proposed approach will yield mechanistic insights into MOR-initiated signaling pathways and how these are regulated by receptor trafficking. Identifying key regulators of MOR activation will fill a critical gap for designing safer, pathway selective analgesics and treatments for opioid addiction.

概括阿片类药物是最有效的镇痛药，但与严重的副作用有关，包括呼吸抑郁，容忍和成瘾。这些因素有助于导致美国阿片类药物滥用流行，使在美国，药物过量导致意外死亡的主要原因。那，鉴定更安全的镇痛药副作用减少和滥用潜力对于解决持续的危机至关重要。临床使用的阿片类药物主要通过对µ-阿片受体（MOR）的作用发挥其镇痛作用和不良反应。虽然采用了几种方法用于安全镇痛药，但这些努力受到缺乏的限制了解MOR参与和激活配体结合的复杂生化网络。该提议建立在最近的证据的基础上，表明（1）MOR信令取决于亚细胞定位和以配体特异性方式进行的膜运输，（2）MOR显示配体对其蛋白质相互作用网络及其激活的信号通路的依赖性影响。那是描绘的内源性肽和添加剂阿片类药物的MOR引发的信号通路以及它们如何由接收者的位置和贩运协调为治疗方式提供了潜在的新策略更安全的镇痛药。该提案的总体目标是结合定量蛋白质组学基因组学和阿片受体生物学系统地发现和表征MOR信号的调节剂并贩运人类引起的多能干细胞衍生的神经元。我们将结合接近标签质量光谱和定量磷蛋白质组学系统地描绘了相互作用网络参与并绘制其激活的信号通路。研究蛋白质组学靶标在MOR中的功能作用信号和贩运，我们将开发和应用记者测定法进行接收器信号和贩运 CRISPRI基因调节筛选。最后，我们将测试蛋白质组学和遗传学的机械假设屏幕介绍了新颖的运输和信号调节器如何微调MOR激活的细胞反应。我们提出的方法将产生对MOR引发的信号通路以及它们的机械见解由接收器贩运监管。识别MOR激活的关键调节器将填补设计的关键空白更安全的途径选择性镇痛药和阿片类药物成瘾的治疗方法。