Genetically-Targeted Photo-Pharmacology for Native Opioid Receptors

天然阿片受体的基因靶向光药理学

基本信息

批准号：
10397653
负责人：
Joshua Levitz
金额：
$ 45.03万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10397653
关键词：
Acute Agonist Analgesics Antibodies Antidepressive Agents Behavior Behavioral Behavioral Assay Binding Biology Brain Brain region Calcium Cells Chemicals Chronic stress Complex Conflict (Psychology)Coupling Dangerousness Dependence Development Disease Disinhibition Drug Addiction Drug Targeting Electrophysiology (science)Engineering Family Fentanyl Functional disorder G-Protein-Coupled Receptors GTP-Binding Proteins Genes Image In Vitro Interneurons Label Ligands Measures Medial Mediating Mental Depression Mental disorders Methodology Methods Modification Molecular Molecular Probes Mood Disorders Moods Morphine Mus N-terminal Naloxone Nervous system structure Neuromodulator Neurons Opioid Opioid Receptor Opioid agonist Optics Pain management Pathway interactions Peptide Signal Sequences Perception Pharmaceutical Preparations Pharmacology Pharmacology Study Phase Prefrontal Cortex Process Receptor Activation Rewards Role S-nitro-N-acetylpenicillamine Sampling Sensory Process Signal Transduction Slice Somatostatin Specificity Structure Synapses System Techniques Testing Therapeutic Time Work addiction antagonist antidepressant effect azobenzene base beta-arrestin brain cell cell type chemical synthesis desensitization design expectation experimental study improved in vivo insight interest molecular dynamics mood regulation mu opioid receptors nanobodies nervous system disorder neural circuit neuronal excitability neuroregulation pain sensation photoactivation rational design receptor receptor internalization reward processing side effect small molecule spatiotemporal targeted treatment temporal measurement two-photon

项目摘要

PROJECT SUMMARY In the nervous system, G protein-coupled receptors (GPCRs) sense neuromodulators to initiate intracellular signaling cascades that control a plethora of brain functions. Of particular importance are the opioid receptors which contribute to pain sensation, reward processing and mood regulation. Consistent with these roles, opioid receptors serve as major drug targets for a variety of disorders. Agonists of the mu-opioid receptor (MOR) are common analgesics and also have potential as antidepressants. However, a challenge with opioid- based treatment is the propensity for addiction, tolerance and dangerous side effects. Unfortunately, limitations in the precision of pharmacological approaches have hampered our ability to dissect the molecular, cellular and circuit level mechanisms of MOR-mediated disease treatment and develop improved therapeutic strategies. We previously established methodologies for optical control of GPCRs using photopharmacology, which we will adapt for the MOR in the R61 phase. In aim 1, we will develop a range of new photoswitchable compounds for the MOR using a combination of structure-based prediction, chemical synthesis and functional analysis. In aim 2, we will adapt these compounds to make photoswitchable orthogonal remotely-tethered ligands (PORTLs) which covalently attach to target receptors with a labeling tag, such as SNAP, CLIP or Halo. To enable targeting of native receptors, we will further extend our system to develop nanobody-photoswitch conjugates (NPCs) which bind to native receptors and deliver a PORTL for reversible optical control. NPCs can either be genetically- encoded to permit cell type-targeting or can be purified in vitro and directly applied to the sample in a gene-free approach. Together, this will provide a new, widely-applicable toolset for MORs while also developing an engineering framework for extension of this approach across different receptor types. In the R33 phase, we will harness our toolset in vivo to probe the basis of MOR-mediated antidepressant effects in mice (aim 3). We hypothesize that activation of MORs localized on interneurons of the medial prefrontal cortex (mPFC) induce disinhibition that leads to normalization of dysfunctional prefrontal circuits. PORTLs and NPCs will enable targeting to the cell types and brain regions of interest with sufficient spatiotemporal precision to probe the relationship between receptor activation and behavioral modulation. We will use behavioral assays of relevance to depression, as well as measures of reward and dependence with the expectation that targeted photo-activation can produce antidepressant effects with minimal side effects. To gain further mechanistic insight, we will perform slice electrophysiology and in vivo 2-photon calcium imaging to measure the effects of MOR activation on mPFC activity. This work will validate our toolset in vivo and provide a key step toward understanding the mechanism of MOR modulation for depression treatment.

项目摘要在神经系统中，G蛋白偶联受体（GPCR）感觉神经调节剂启动控制大脑功能的细胞内信号传导级联。阿片类药物特别重要有助于疼痛感，奖励加工和情绪调节的受体。与这些一致角色，阿片受体是多种疾病的主要药物靶标。 mu阿片受体的激动剂（MOR）是常见的镇痛药，也具有抗抑郁药的潜力。但是，阿片类药物的挑战 - 基于治疗是成瘾，耐受性和危险副作用的倾向。不幸的是，局限性从药理学方法的精确度中，我们妨碍了我们剖析分子，细胞和 MOR介导的疾病治疗的电路水平机制，并发展了改进的治疗策略。我们以前使用光肢体学对GPCR的光学控制建立了方法论，我们将在R61阶段适应MOR。在AIM 1中，我们将开发一系列新的可拍打化合物使用基于结构的预测，化学合成和功能分析的组合。在 AIM 2，我们将适应这些化合物，以使照片开关的正交遥控配体（portls）共价连接到具有标签标签的目标受体上，例如快照，夹子或光环。启用定位在天然受体中，我们将进一步扩展我们的系统以开发纳米型 - 光旋转偶联物（NPC）与天然受体结合并提供可逆光控制的端口。 NPC可以是遗传的编码以允许细胞类型靶向或可以在体外纯化并直接应用于无基因的样品方法。一起，这将为MOR提供新的，广泛的可应用工具集，同时还为用于扩展这种方法跨不同受体类型的工程框架。在R33阶段，我们将在体内利用我们的工具集来探测MOR介导的抗抑郁药的基础在小鼠中的影响（AIM 3）。我们假设在内侧前额叶中间神经元上的MOR激活皮层（MPFC）引起的抑制作用，导致前额叶功能障碍的正常化。端口和 NPC将以足够的时空精度为目标靶向感兴趣的细胞类型和大脑区域探测受体激活与行为调制之间的关系。我们将使用行为分析与抑郁症相关，以及奖励和依赖的度量与目标的期望光激活可以产生抗抑郁作用，其副作用最少。获得进一步的机械洞察力，我们将执行切片的电生理学和体内2-光子钙成像，以测量 MOR激活MPFC活性。这项工作将在体内验证我们的工具集，并为朝着了解MOR调制抑郁治疗的机制。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Bioinspired Synthesis of (-)-PF-1018.

DOI：
10.1002/anie.201912452
发表时间：
2020-03-23
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Quintela-Varela, Hugo;Jamieson, Cooper S.;Shao, Qianzhen;Houk, K. N.;Trauner, Dirk
通讯作者：
Trauner, Dirk