J Oral Facial Pain Headache. 2016 Winter;30(1):42-50. doi: 10.11607/ofph.1512.
To determine whether herpes simplex virus-based vectors can efficiently transduce mouse trigeminal ganglion (TG) neurons and attenuate preexisting nerve injury-induced whisker pad mechanical hypersensitivity in a trigeminal inflammatory compression (TIC) neuropathic pain model.
Tissue transduction efficiencies of replication-conditional and replication-defective vectors to mouse whisker pads after topical administration and subcutaneous injection were assessed using quantitative real-time PCR (qPCR). Tissue tropism and transgene expression were assessed using qPCR and reverse-transcriptase qPCR following topical application of the vectors. Whisker pad mechanical sensitivities of TIC-injured mice were determined using graduated von Frey fibers before and after application of human preproenkephalin expressing replication-conditional vector (KHPE). Data were analyzed using one-way analysis of variance (ANOVA) and post hoc tests.
Transduction of target TGs was 8- to 50-fold greater after topical application than subcutaneous injection and ≥ 100-fold greater for replication-conditional than replication-defective vectors. Mean KHPE loads remained constant in TGs (4.5-9.8 × 10(4) copies/TG) over 3 weeks but were below quantifiable levels (10 copies/tissue) within 2 weeks of application in other nontarget cephalic tissues examined. Transgene expression in TGs was maximal during 2 weeks after topical application (100-200 cDNA copies/mL) and was below quantifiable levels (1 cDNA copy/mL) in all nontarget tissues. Topical KHPE administration reduced TIC-related mechanical hypersensitivity on whisker pads 4-fold (P < .05) for at least 1 week.
Topically administered KHPE produced a significant antinociceptive effect in the TIC mouse model of chronic facial neuropathic pain. This is the first report in which a gene therapeutic approach reduced trigeminal pain-related behaviors in an established pain state in mice.
确定单纯疱疹病毒载体是否能有效地转导小鼠三叉神经节 (TG) 神经元，并减轻三叉神经炎性压迫 (TIC) 神经病理性疼痛模型中预先存在的神经损伤诱导的胡须垫机械性超敏反应。
使用定量实时 PCR (qPCR) 评估复制条件和复制缺陷载体在小鼠胡须垫经皮给药和皮下注射后的组织转导效率。使用 qPCR 和逆转录 qPCR 评估载体经皮应用后的组织嗜性和转基因表达。在应用表达人前脑啡肽原的复制条件载体 (KHPE) 前后，使用分级冯弗雷纤维测定 TIC 损伤小鼠的胡须垫机械敏感性。使用单向方差分析 (ANOVA) 和事后检验分析数据。
与皮下注射相比，经皮应用后对 TG 的转导增加了 8 到 50 倍，与复制缺陷载体相比，复制条件载体的转导增加了 100 倍以上。在 3 周内，KHPE 负荷在 TG 中保持不变（4.5-9.8×10(4)拷贝/TG），但在应用后 2 周内，在检查的其他非目标头颈部组织中低于可定量水平（10 拷贝/组织）。在 TG 中，转基因表达在经皮应用后 2 周内达到最大值（100-200 cDNA 拷贝/mL），在所有非目标组织中均低于可定量水平（1 cDNA 拷贝/mL）。KHPE 经皮给药可使 TIC 相关机械性超敏反应减少 4 倍（P<.05），至少持续 1 周。
在慢性面部神经病理性疼痛的 TIC 小鼠模型中，经皮给予 KHPE 产生了显著的镇痛作用。这是首例报道基因治疗方法在小鼠的既定疼痛状态下减轻三叉神经疼痛相关行为的研究。
