OPIOID RECEPTOR GENE TRANSFER-PAIN AND TOLERANCE

阿片受体基因转移-疼痛和耐受性

• 批准号: 6405267
• 负责人: LI-YEN M HUANG
• 金额: $ 36.13万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-25 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6405267
• 关键词: Freund's adjuvant; adeno associated virus group; analgesia; analgesics; biotechnology; chronic pain; gene delivery system; gene therapy; immunocytochemistry; immunofluorescence technique; laboratory rat; neurogenetics; neurons; nonhuman therapy evaluation; opioid receptor; pain threshold; receptor binding; spinal ganglion; statistics /biometry; transfection /expression vector

The long-term goal of this research is to develop gene therapy for chronic pain. Opiates have remained to be the drugs of choice for treating patients with severe pain. The side effects of opiates, including respiratory depression, tolerance and dependence, have often limited their use. The focus of this grant is to design viral vectors that allow for efficient and stable delivery of wild type or mutant mu opioid receptor (muOR) genes to sensory neurons. We hypothesize that appropriate muOR gene delivery and expression in dorsal root ganglion (DRG) neurons will lower the doses of mu-opioids required for analgesia and reduce tolerance, thus decreasing the side effects of opioids. To test this hypothesis, we will (1) deliver the wild type or mutant mu OR gene into sensory neurons using an adeno- associated virus (AAV) vector, (2) evaluate the effects of mu- opioids on pain behaviors in inflamed rats injected with recombinant AAV-mediated muOR transgene (rAAV-muOR), (3) evaluate the effects of mu-opioid in tolerant rats that receive injections of rAAV-muOR and (4) evaluate the effects of mu-opioids on membrane conductance and receptor trafficking in rAAV-muOR- transduced DRG neurons isolated from non tolerant and tolerant rats. The experiments will be performed on rats treated with complete Freund's adjuvant (CFA) and on DRG neurons isolated from those rats. Pain behaviors will be monitored by paw withdrawal latencies, membrane currents will be measured with perforated patch electrodes and receptor trafficking will be examined with immunofluorescence staining and receptor binding assays. These experiments will establish the feasibility of a genetic approach to pain treatment. If successful, the study will provide clinicians with a new tool to treat patients with severe pain and provide researchers a model for studying the role of opioid receptor regulation in the development of opioid tolerance.

这项研究的长期目标是开发针对慢性疼痛的基因疗法。 阿片类药物仍然是治疗剧烈疼痛患者的首选药物。 阿片类药物的副作用，包括呼吸抑制、耐受性和依赖性，往往限制了其使用。 该资助的重点是设计病毒载体，允许将野生型或突变型 mu 阿片受体 (muOR) 基因有效且稳定地递送至感觉神经元。 我们假设，在背根神经节（DRG）神经元中适当的 muOR 基因传递和表达将降低镇痛所需的 mu-阿片类药物的剂量并降低耐受性，从而减少阿片类药物的副作用。 为了检验这一假设，我们将（1）使用腺相关病毒（AAV）载体将野生型或突变型 mu OR 基因传递到感觉神经元中，（2）评估 mu-阿片类药物对注射重组 AAV 介导的 muOR 转基因（rAAV-muOR）的发炎大鼠疼痛行为的影响，（3）评估 mu-阿片类药物对耐受大鼠的影响 (4) 评估 mu-阿片类药物对从非耐受和耐受大鼠分离的 rAAV-muOR 转导的 DRG 神经元的膜电导和受体运输的影响。 实验将在用完全弗氏佐剂（CFA）治疗的大鼠和从这些大鼠中分离的 DRG 神经元上进行。 将通过缩爪潜伏期监测疼痛行为，用穿孔贴片电极测量膜电流，并用免疫荧光染色和受体结合测定检查受体运输。这些实验将确立基因方法治疗疼痛的可行性。 如果成功，该研究将为临床医生提供治疗剧烈疼痛患者的新工具，并为研究人员提供研究阿片受体调节在阿片耐受性发展中作用的模型。