Targeting GPCRs in amygdalar and cortical neural ensembles to treat pain aversion

靶向杏仁核和皮质神经群中的 GPCR 来治疗疼痛厌恶

• 批准号: 10055582
• 负责人: Gregory Scherrer
• 金额: $ 280.29万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10055582
• 关键词: Acute Pain; Affect; Affective; Agonist; American; Amygdaloid structure; Analgesics; Animal Model; Animals; Anterior; Anxiety; Area; Behavior; Behavioral Assay; Bioinformatics; Brain; Breathing; Catalogs; Cells; Clinical; Code; Collaborations; Complement; Coupled; Disease; Drug Modulation; Drug Targeting; Electrophysiology (science); Emotional; Emotions; Fluorescence-Activated Cell Sorting; Fluorescent in Situ Hybridization; G-Protein-Coupled Receptors; Genetic; Genomics; Human; In Situ Hybridization; Label; Lateral; Light; Mechanics; Mediating; Mental Depression; Methods; Molecular; Monitor; Mus; Neurons; Nociception; Opioid; Opioid Analgesics; Opioid Receptor; Pain; Pain quality; Pathway interactions; Patients; Pattern; Peripheral nerve injury; Pharmaceutical Preparations; Population; Property; Proteins; Pyramidal Cells; RNA; Reflex action; Reporting; Research; Rewards; Rodent; Role; SSTR2 gene; Safety; Science; Sensory; Shapes; Slice; Somatostatin Receptor; Stimulus; Testing; Tracer; Translating; Validation; Viral; Whole Body Plethysmography; Wild Type Mouse; Withdrawal; allodynia; base; brain tissue; cell cortex; chronic pain; chronic pain patient; cingulate cortex; cingulotomy; comorbidity; designer receptors exclusively activated by designer drugs; dosage; experience; hippocampal pyramidal neuron; human tissue; imaging study; in vivo; in vivo imaging; mouse genetics; negative affect; neural circuit; neurophysiology; novel; novel therapeutics; pain patient; pain relief; painful neuropathy; promoter; receptor; relating to nervous system; response; reward processing; side effect; single-cell RNA sequencing; spontaneous pain

PROJECT SUMMARY Pain is a multidimensional experience with sensory and affective components. The aversive quality of pain, i.e. its inherent unpleasantness, causes a majority of chronic pain patients’ suffering and often leads to comorbid disorders such as anxiety and depression. Despite their addictive qualities, opioid analgesics remain clinically useful since they can profoundly dampen pain affect. Thus, discovering targets that could alter neural activity selectively in neural circuits that generate pain aversion, but not in the reward or breathing circuits that opioids also alter, is an attractive strategy to develop novel, safer analgesics. Recently, by combining in vivo imaging and chemogenetic manipulations of neural dynamics in the basal and lateral amygdala (BLA) of freely behaving mice encountering noxious stimuli, our collaboration discovered a distinct neural ensemble in the BLA that encodes the negative affective valence of pain (Corder et al., Science, 2019). Chemogenetic inhibition of this nociceptive coding ensemble using Gi/o-protein-coupled-DREADDs alleviated pain affective behaviors without altering withdrawal reflexes, anxiety or reward. Moreover, our functional studies of this nociceptive ensemble revealed its causal role in the phenomenon of allodynia. Based on these exciting findings, we now seek to identify novel targets to treat pain by determining the molecular identity of these BLA nociceptive cells via in situ hybridization and single cell RNA-sequencing (scRNA-seq). Our preliminary scRNA-seq studies of BLA nociceptive cells suggest they express dozens of Gi/o protein-coupled receptors (Gi/o-GPCRs) that could be targeted for anti-nociception against pain affect. Further, our tracing studies have revealed a set of layer V pyramidal cells in anterior cingulate cortex (ACC) that project onto BLA nociceptive neurons, consistent with the fact cingulotomy can be used to treat intractable chronic pain. Resolving the molecular identity of these ACC nociceptive cells could also reveal new targets to treat pain affect. Thus, here we propose to catalog candidate Gi/o-GPCR targets in BLA and ACC (Aim 1, Discovery), test their utility to treat pain (Aim 2, Validation), and verify these new targets have no effect in the brain’s reward and breathing circuitry (Aim 3, Safety & Translatability). In Aim 1 we will identify Gi/o-GPCR targets in pain affect circuits of the BLA and ACC using mouse genetics, viral tracers, scRNA-seq and bioinformatics analyses. In Aim 2, we will validate the neurophysiological effects and analgesic properties of these new targets, using electrophysiological recordings in live brain tissue slices, animal models of acute and chronic pain, and Ca2+ imaging studies in behaving mice of BLA and ACC neural activity. In Aim 3, we will verify the safety and translatability of the novel antinociceptive drug targets. We will evaluate each target for abuse potential and effects on breathing by using behavioral assays for reward processing and whole-body plethysmography, respectively. To evaluate whether our results in rodents are likely to translate clinically, we will also analyze expression patterns of the drug targets in human tissue using in situ hybridization.

项目摘要 疼痛是一种具有感官和情感成分的多维体验。痛苦的厌恶质量，即 它固有的不愉快性会引起大多数慢性疼痛患者的痛苦，并且经常导致合并症 焦虑和抑郁等疾病。尽管具有附加品质，但阿片类镇痛药在临床上仍然 很有用，因为它们会深深地影响疼痛。那是发现可以改变神经活动的靶标 在神经元电路中有选择地产生疼痛厌恶，但在阿片类药物的奖励或呼吸电路中没有 另外，Alter是开发新颖，更安全的镇痛药的有吸引力的策略。 最近，通过结合基本神经动力学的体内成像和化学发生操作 和自由行为遇到有害刺激的自由行为的侧向杏仁核（BLA），我们发现的合作已发现 BLA中独特的神经合奏，编码疼痛的负面情感价（Corder等，科学， 2019）。使用gi/o蛋白偶联的划线对这种伤害感受器编码集合的化学抑制作用 缓解疼痛情感行为而不改变戒断反射，焦虑或奖励。而且，我们的 对这种伤害性集成的功能研究揭示了其在异肌现象中的因果作用。 基于这些令人兴奋的发现，我们现在试图通过确定来确定新的目标来治疗疼痛 这些BLA伤害感受细胞通过原位杂交和单细胞RNA测序的分子身份 （scrna-seq）。我们对BLA伤心细胞的初步SCRNA-SEQ研究表明它们表达了数十个GI/O 蛋白质偶联受体（GI/O-GPCR）可以针对疼痛影响抗伤害感受。此外， 我们的追踪研究揭示了该项目的前扣带回皮层（ACC）中的一组V层锥体细胞 在BLA伤害性神经元上，与扣可毛术一致的事实可用于治疗棘手的慢性疼痛。 解决这些ACC伤害性细胞的分子身份也可能揭示出治疗疼痛影响的新靶标。 这是，我们在这里建议在BLA和ACC（AIM 1，Discovery）中对候选GI/O-GPCR目标进行分类 他们治疗疼痛的效用（AIM 2，验证）并验证这些新目标对大脑的奖励没有影响，并且 呼吸电路（AIM 3，安全性和转换性）。在AIM 1中，我们将确定疼痛影响中的GI/O-GPCR目标 使用小鼠遗传学，病毒示踪剂，SCRNA-SEQ和生物信息学分析的BLA和ACC电路。在 AIM 2，我们将使用这些新靶标的神经生理效应和镇痛特性 活脑组织切片中的电生理记录，急性和慢性疼痛的动物模型以及Ca2+ 在BLA和ACC神经活动的行为小鼠中进行成像研究。在AIM 3中，我们将验证安全性和 新型抗伤害性药物靶标的翻译性。我们将评估每个目标的滥用潜力， 通过使用行为测定进行奖励处理和全身多生产描记的影响，对呼吸的影响， 分别。为了评估我们在啮齿动物中的结果是否可能在临床上翻译，我们还将分析 使用原位杂交在人体组织中药物靶标的表达模式。