Xu Ya, Gu Yanping, Wu Ping, Li Guang-Wen, Huang Li-Yen Mae
Marine Biomedical Institute, University of Texas Medical Branch, Galveston, TX 77555, USA.
Hum Gene Ther. 2003 Jun 10;14(9):897-906. doi: 10.1089/104303403765701187.
Transferring therapeutic genes into the nociceptive system, including dorsal root ganglia (DRGs) and the spinal cord, is potentially a powerful approach for the treatment of chronic pain in humans. Adeno-associated viral vectors (AAVs) are particularly useful in delivering foreign genes to targeted tissues because they seldom induce immune responses or produce cytotoxicity. To determine the efficiency of transgene expression and the best route(s) of delivery, a recombinant AAV type 2 vector containing the enhanced green fluorescent protein (EGFP) gene driven by the neuron-specific enolase (NSE) promoter (rAAV-EGFP) was constructed. We injected the vector into subcutaneous tissue, sciatic nerve, DRGs, and subarachnoid space, and examined EGFP expression in the DRG, spinal cord, and nerve fibers. Both sciatic nerve and DRG injection led to strong EGFP expression in a large number of DRG neurons. The expression persisted for more than 6-8 months. We then delivered the mu-opioid receptor (muOR) gene into DRGs through direct DRG or sciatic nerve injection of rAAV-muOR and found a significant increase in morphine efficacy. These results suggest that delivering therapeutic genes to DRGs by the rAAV-NSE vector is a valid strategy for treatment of chronic pain.
将治疗性基因导入伤害感受系统，包括背根神经节（DRG）和脊髓，可能是治疗人类慢性疼痛的一种有效方法。腺相关病毒载体（AAV）在将外源基因传递到靶向组织方面特别有用，因为它们很少诱导免疫反应或产生细胞毒性。为了确定转基因表达的效率和最佳递送途径，构建了一种重组2型腺相关病毒载体，其包含由神经元特异性烯醇化酶（NSE）启动子驱动的增强型绿色荧光蛋白（EGFP）基因（rAAV-EGFP）。我们将该载体注射到皮下组织、坐骨神经、背根神经节和蛛网膜下腔，并检测背根神经节、脊髓和神经纤维中的EGFP表达。坐骨神经和背根神经节注射均导致大量背根神经节神经元中强烈的EGFP表达。这种表达持续超过6-8个月。然后，我们通过直接向背根神经节或坐骨神经注射rAAV-μOR将μ-阿片受体（μOR）基因导入背根神经节，发现吗啡疗效显著提高。这些结果表明，通过rAAV-NSE载体将治疗性基因递送至背根神经节是治疗慢性疼痛的一种有效策略。
