Effects of spinal non-viral interleukin-10 gene therapy formulated with d-mannose in neuropathic interleukin-10 deficient mice: Behavioral characterization, mRNA and protein analysis in pain relevant tissues.
Effects of spinal non-viral interleukin-10 gene therapy formulated with d-mannose in neuropathic interleukin-10 deficient mice: Behavioral characterization, mRNA and protein analysis in pain relevant tissues.
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DOI：
10.1016/j.bbi.2017.11.004
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发表时间：
2018-03
期刊：
影响因子：
--
通讯作者：
Milligan ED
中科院分区：
文献类型：
--
作者：
Vanderwall AG;Noor S;Sun MS;Sanchez JE;Yang XO;Jantzie LL;Mellios N;Milligan ED
研究表明，脊髓(鞘内；I.T.)白介素10(IL-10)基因治疗可逆转神经病理性疼痛的动物模型，与甘露糖受体(MR；CD206)配体D-甘露糖(DM)联合应用可显著提高治疗效果。然而，内源性IL-10的作用可能是在I.T.后观察到的持久疼痛控制所必需的。IL-10基因治疗，潜在地缩小了这种非病毒转基因传递方法的应用范围。在这里，我们展示了IT。应用表达IL-10转基因的裸质粒DNA与DM(DM/PDNA-IL-10)共注射治疗IL-10缺陷(IL-10 KO)小鼠的周围神经病理性疼痛，可产生深刻而持久的双侧疼痛抑制。单侧坐骨神经慢性缩窄性损伤(CCI)诱发神经病理性疼痛，经DM/PDNA-IL-10联合治疗后，WT和IL-10KO小鼠的光触觉敏感性(机械性超敏)均持续缓解，而I.T后可一过性逆转。只有DM一个人。在给予DM/PDNA-IL-10的稳定缓解疼痛的IL-10KO小鼠中，IL-10转基因基因的mRNA在马尾神经和同侧背根神经节(DRG)中检测到，但在腰髓中没有检测到。此外，应用DM/pDNA-IL-10可增加同侧背根神经节的抗炎转化生长因子-β-1，降低促炎的肿瘤坏死因子m RNA。此外，DM/PDNA-IL-10处理的小鼠表现出脊髓促炎因子TNF、CCL2(MCP-1)和小胶质细胞特异性标记物TMEM119的mRNA表达减少。类似地，DM/PDNA-IL-10治疗降低了腰椎脊髓背角星形胶质细胞激活标记GFAP的免疫反应。尽管DM治疗的小鼠出现一过性逆转和早期恢复痛觉异常，但腰髓显示出高水平的肿瘤坏死因子、CCL2和TMEM119mRNA水平。在CCI操作后，DRG中MR(CD206)和IL-10受体mRNAs的表达增加，而不受注射治疗的影响，这表明病理条件刺激了DM/PDNA-IL-10联合治疗所需的关键解剖区域相关受体的上调和可用性。综上所述，目前的报告表明，非病毒性DM/PDNA-IL-10基因治疗不需要内源性IL-10来持久缓解周围神经病理性疼痛，也不需要直接接触脊髓背角来强劲和持久地缓解神经病理性疼痛。脊髓非病毒DM/PDNA-IL-10联合治疗可能为发展其他中枢神经系统疾病的非病毒基因治疗方法提供一个框架。
Studies show that spinal (intrathecal; i.t.) interleukin-10 (IL-10) gene therapy reverses neuropathic pain in animal models, and co-administration with the mannose receptor (MR; CD206) ligand D-mannose (DM) greatly improves therapeutic efficacy. However, the actions of endogenous IL-10 may be required for enduring pain control observed following i.t. IL-10 gene therapy, potentially narrowing the application of this non-viral transgene delivery approach. Here, we show that i.t. application of naked plasmid DNA expressing the IL-10 transgene co-injected with DM (DM/pDNA-IL-10) for the treatment of peripheral neuropathic pain in IL-10 deficient (IL-10 KO) mice results in a profound and prolonged bilateral pain suppression. Neuropathic pain is induced by unilateral sciatic chronic constriction injury (CCI), and while enduring relief of light touch sensitivity (mechanical allodynia) in both WT and IL-10 KO mice was observed following DM/pDNA-IL-10 co-therapy, transient reversal from allodynia was observed following i.t. DM alone. In stably pain-relieved IL-10 KO mice given DM/pDNA-IL-10, mRNA for the IL-10 transgene is detected in the cauda equina and ipsilateral dorsal root ganglia (DRG), but not the lumbar spinal cord. Further, DM/pDNA-IL-10 application increases anti-inflammatory TGF-β1 and decreases pro-inflammatory TNF mRNA in the ipsilateral DRG compared to allodynic controls. Additionally, DM/pDNA-IL-10 treated mice exhibit decreased spinal pro-inflammatory mRNA expression for TNF, CCL2 (MCP-1), and for the microglial-specific marker TMEM119. Similarly, DM/pDNA-IL-10 treatment decreases immunoreactivity for the astrocyte activation marker GFAP in lumbar spinal cord dorsal horn. Despite transient reversal and early return to allodynia in DM-treated mice, lumbar spinal cord revealed elevated TNF, CCL2 and TMEM119 mRNA levels. Both MR (CD206) and IL-10 receptor mRNAs are increased in the DRG following CCI manipulation independent of injection treatment, suggesting that pathological conditions stimulate upregulation and availability of relevant receptors in critical anatomical regions required for the therapeutic actions of the DM/pDNA-IL-10 co-therapy. Taken together, the current report demonstrates that non-viral DM/pDNA-IL-10 gene therapy does not require endogenous IL-10 for enduring relief of peripheral neuropathic pain and does not require direct contact with the spinal cord dorsal horn for robust and enduring relief of neuropathic pain. Spinal non-viral DM/pDNA-IL-10 co-therapy may offer a framework for the development of non-viral gene therapeutic approaches for other diseases of the central nervous system.
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影响因子：
4.1
作者：
Garcia-Aguilar T;Espinosa-Cueto P;Magallanes-Puebla A;Mancilla R
通讯作者：
Mancilla R
影响因子：
3.3
作者：
Echeverry S;Shi XQ;Haw A;Liu H;Zhang ZW;Zhang J
通讯作者：
Zhang J
影响因子：
3.3
作者：
FITZGERALD, M;WOOLF, CJ
通讯作者：
WOOLF, CJ
影响因子：
2.9
作者：
FITZGERALD, M
通讯作者：
FITZGERALD, M
影响因子：
5.3
作者：
Colburn, RW;Rickman, AJ;DeLeo, JA
通讯作者：
DeLeo, JA