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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，但会带来严重的副作用，包括呼吸系统疾病抑郁、耐受和成瘾。这些因素导致了美国阿片类药物滥用的流行，使得药物过量是美国意外死亡的主要原因。因此，确定更安全的镇痛药减少副作用和滥用可能性对于解决当前危机至关重要。临床上使用的阿片类药物主要通过作用于μ-阿片受体（MOR）来发挥镇痛和不良作用。尽管采取了多种方法来获得更安全的镇痛药，但这些努力因缺乏了解 MOR 响应配体结合而参与和激活的复杂生化网络。该提议建立在最近的证据之上，表明 (1) MOR 信号传导取决于之间的相互作用以配体特异性方式进行亚细胞定位和膜运输，并且 (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影响。由此，划定 MOR 启动的内源性肽和成瘾性阿片类药物的信号传导途径以及它们是如何发生的通过受体位置和贩运进行协调，为治疗方式和方法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将定量蛋白质组学、功能基因组学和阿片受体生物学，系统地发现和表征 MOR 信号调节因子以及人类诱导多能干细胞衍生神经元的贩运。我们将结合接近标记质量光谱测定法和定量磷酸化蛋白质组学系统地描绘了 MOR 的相互作用网络参与并绘制其激活的信号通路。研究蛋白质组靶标在 MOR 中的功能作用信号传导和贩运，我们将开发并应用受体信号传导和贩运的报告基因检测 CRISPRi 基因调控筛选。最后，我们将测试蛋白质组学和遗传学的机制假设筛选新的运输和信号调节因子如何微调 MOR 激活的细胞反应。我们提出的方法将产生对 MOR 启动的信号通路及其如何发生的机制见解受受体运输调节。确定 MOR 激活的关键调节因子将填补设计的关键空白更安全的途径选择性镇痛药和治疗阿片类药物成瘾的方法。