The Alpha Isoform of Heat Shock Protein 90 and the Co-chaperones p23 and Cdc37 Promote Opioid Anti-nociception in the Brain
The Alpha Isoform of Heat Shock Protein 90 and the Co-chaperones p23 and Cdc37 Promote Opioid Anti-nociception in the Brain
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DOI：
10.3389/fnmol.2019.00294
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发表时间：
2019-11-29
影响因子：
4.8
通讯作者：
Streicher, John M.
中科院分区：
文献类型：
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作者：
Lei, Wei;Duron, David, I;Streicher, John M.
μ阿片受体（莫尔）的阿片样物质活化促进信号级联，其引起对疼痛的镇痛反应和副作用如成瘾和依赖。操纵这些级联反应，例如通过偏性激动，有很大的希望改善阿片类药物治疗。然而，莫尔的信号级联通常知之甚少，为药物开发提供了很少的靶点。在我们早期的工作中，我们确定热休克蛋白90（Hsp 90）作为一种新的和重要的调节阿片类药物的抗伤害性在大脑中通过促进ERK MAPK激活。在这项研究中，我们试图确定Hsp 90的分子异构体和辅助分子伴侣，这可能为未来的临床干预提供特异性靶点。我们使用新型选择性小分子抑制剂以及通过脑室内（icv）途径递送至成年CD-1小鼠大脑的CRISPR/Cas9基因编辑构建体，以靶向Hsp 90亚型（Hsp 90 alpha/beta，Grp 94）和辅助分子伴侣（p23，Cdc 37，Aha 1）。我们发现，在术后爪切口疼痛模型中，抑制Hsp 90 α亚型完全阻断了吗啡的抗伤害性感受，同时阻断了ERK和JNK MAPK的激活，这表明Hsp 90 α是脑中阿片样物质反应的主要调节剂。我们进一步发现，共伴侣蛋白p23和Cdc 37的抑制阻断了吗啡的抗伤害感受，这表明这些共伴侣蛋白有助于Hsp 90 α促进阿片类药物的抗伤害感受。最后，我们在大脑中使用放线菌酮治疗来证明阿片类药物治疗30分钟内的快速蛋白质翻译是Hsp 90调节阿片类药物反应所必需的。这些研究共同提供了对Hsp 90促进阿片类抗伤害感受的分子机制的深入了解。因此，这些发现既提高了我们对莫尔信号转导的基础科学知识，也可以为临床干预提供未来的目标，以改善阿片类药物治疗。
Opioid activation of the mu opioid receptor (MOR) promotes signaling cascades that evoke both analgesic responses to pain and side effects like addiction and dependence. Manipulation of these cascades, such as by biased agonism, has great promise to improve opioid therapy. However, the signaling cascades of the MOR are in general poorly understood, providing few targets for drug development. In our earlier work, we identified Heat shock protein 90 (Hsp90) as a novel and crucial regulator of opioid anti-nociception in the brain by promoting ERK MAPK activation. In this study, we sought to identify the molecular isoforms and co-chaperones by which Hsp90 carried out this role, which could provide specific targets for future clinical intervention. We used novel selective small molecule inhibitors as well as CRISPR/Cas9 gene editing constructs delivered by the intracerebroventricular (icv) route to the brains of adult CD-1 mice to target Hsp90 isoforms (Hsp90 alpha/beta, Grp94) and co-chaperones (p23, Cdc37, Aha1). We found that inhibition of the isoform Hsp90 alpha fully blocked morphine anti-nociception in a model of post-surgical paw incision pain, while blocking ERK and JNK MAPK activation, suggesting Hsp90 alpha as the main regulator of opioid response in the brain. We further found that inhibition of the co-chaperones p23 and Cdc37 blocked morphine anti-nociception, suggesting that these co-chaperones assist Hsp90 alpha in promoting opioid anti-nociception. Lastly, we used cycloheximide treatment in the brain to demonstrate that rapid protein translation within 30 min of opioid treatment is required for Hsp90 regulation of opioid response. Together these studies provide insight into the molecular mechanisms by which Hsp90 promotes opioid anti-nociception. These findings thus both improve our basic science knowledge of MOR signal transduction and could provide future targets for clinical intervention to improve opioid therapy.