Mechanisms of cannabinoid tolerance

大麻素耐受机制

• 批准号: 10224956
• 负责人: Josee Guindon
• 金额: $ 37.02万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224956
• 关键词: ADRBK2 gene; Acute Pain; Address; Agonist; Analgesics; Animal Model; Behavior assessment; Biochemical; Biochemistry; C-terminal; CNR1 gene; Cancer Patient; Cannabinoids; Cannabis; Cells; Chemicals; Chemotherapy-Oncologic Procedure; Cisplatin; Clinical; Clinical Management; Development; Disadvantaged; G protein coupled receptor kinase; Goals; Grant; Immunoprecipitation; Knockout Mice; Knowledge; Lead; MAPK8 gene; MAPK9 gene; Malignant Neoplasms; Mediating; Medical; Modeling; Molecular; Molecular Mechanisms of Action; Mus; Mutant Strains Mice; Mutate; Mutation; N-terminal; Nausea; Neurons; Pain; Pain management; Pathologic; Pharmacology; Phosphorylation; Phosphorylation Site; Phosphotransferases; Property; Protein Isoforms; Proteomics; Research Project Grants; Resistance; SP600125; Secondary to; Signal Pathway; Signal Transduction; Site; Tail; Testing; Tetrahydrocannabinol; Therapeutic; Thinking; Tissues; Work; alternative treatment; analog; base; cancer pain; chemical genetics; chemotherapy; chronic pain management; clinically relevant; desensitization; expectation; genetic approach; improved; in vivo; inhibitor/antagonist; innovation; insight; kinase inhibitor; knowledge base; marijuana use; mouse model; mutant; novel; pain model; painful neuropathy; pre-clinical; protein protein interaction; response; sex; stable cell line

This study will investigate the mechanisms of cannabinoid tolerance. This objective will be achieved by determining whether cannabinoid tolerance is mediated through agonist-specific mechanisms using a model of chemotherapy-induced neuropathic pain. Our approach will examine tolerance to the anti-allodynic and antinociceptive effects of ∆9-THC, CP55,940, and WIN55,212-2, three cannabinoid agonists with distinct signaling and chemical features. Tolerance to ∆9-THC antinociception in the tail-flick test was eliminated by pre-treatment of S426A/S430A mutants with SP600125, a selective c-Jun N-terminal kinase (JNK) inhibitor suggesting that JNK (SP600125 inhibitor) and GRK/βarrestin2 (S426/S430A mutation) signaling mechanisms coordinate to mediate tolerance to the antinociceptive effect of ∆9-THC. The first objective of this study is to, fully and systematically, test the hypothesis that cannabinoid tolerance is mediated through agonist-specific mechanisms. The second objective is to test the hypothesis that JNK-mediated tolerance for ∆9-THC requires the presence of β−arrestin2. The third objective is to test the hypothesis that β−arrestin2 and JNKs can form protein-protein interactions in vivo. The fourth objective is to test the hypothesis that JNKs can directly phosphorylate CB1 when activated by ∆9-THC using a technologically innovative chemical-genetic approach. The first three hypotheses will be tested in a clinically relevant model of chemotherapy (cisplatin)-induced model of neuropathic pain. The last hypothesis is equally innovative and will provide important information regarding the molecular mechanism of action that is responsible for JNK-mediated ∆9-THC tolerance. The overarching goal of this project is to gain a better understanding of the agonist-specific mechanisms responsible for cannabinoid tolerance that will facilitate the development of long lasting, highly efficacious, and personalized pain therapies.

这项研究将调查大麻素耐受的机制。这个目标将会实现 通过确定大麻素耐受性是否是通过激动剂特异性机制介导的 使用化疗引起的神经性疼痛模型。我们的方法将检查对 Δ9-THC、CP55,940 和 WIN55,212-2 的抗异常疼痛和抗伤害作用，三个 具有独特的信号传导和化学特征的大麻素激动剂。 Δ9-THC 耐受性 通过对 S426A/S430A 突变体进行预处理，消除了甩尾试验中的镇痛作用。 SP600125，一种选择性 c-Jun N 末端激酶 (JNK) 抑制剂，表明 JNK (SP600125 抑制剂）和 GRK/βarrestin2（S426/S430A 突变）信号机制协调介导 对 Δ9-THC 的抗伤害作用的耐受性。本研究的首要目标是充分且 系统地检验大麻素耐受性是通过激动剂特异性介导的假设 机制。第二个目标是检验 JNK 介导的对 Δ9-THC 耐受性的假设 需要 β−arrestin2 的存在。第三个目标是检验 β−arrestin2 和 JNK 可以在体内形成蛋白质-蛋白质相互作用。第四个目标是检验以下假设： 当使用创新技术被 Δ9-THC 激活时，JNK 可以直接磷酸化 CB1 化学遗传学方法。前三个假设将在临床相关模型中进行测试 化疗（顺铂）诱导的神经性疼痛模型。最后一个假设同样成立 具有创新性，将提供有关分子作用机制的重要信息 负责 JNK 介导的 Δ9-THC 耐受性。该项目的总体目标是获得 更好地了解负责大麻素耐受性的激动剂特异性机制 将促进持久、高效和个性化疼痛疗法的开发。