Chen Yun-Hsiang, Hung Tsai-Wei, Wang Yu-Syuan, Bae Eun-Kyung, Wu Kuo-Jen, Wang Yun, Yu Seong-Jin
Center for Neuropsychiatric Research, National Health Research Institutes, Zhunan, Taiwan.
Department of Life Science, Fu-Jen Catholic University, New Taipei City, Taiwan.
Addict Biol. 2025 Aug;30(8):e70073. doi: 10.1111/adb.70073.
Methamphetamine (Meth) is a psychoactive and neurotoxic chemical. Selective antibodies against Meth molecules have been examined for the treatment of Meth abuse through immunization. Antibodies with high affinity for Meth can capture Meth molecules and reduce Meth response. We previously reported that intraperitoneal administration of adeno-associated virus serotype vector serotype 8 carrying Meth-specific monoclonal antibody transgene (AAV8-MethAb, 2.5 × 10 VGC per mouse) induced long-term and stable expression of Meth-antibody in the peripheral. Mice receiving AAV8-MethAb had a lower Meth level in the blood and brain and attenuated Meth-induced locomotor activity after an acute dose of Meth. The effect of AAV-MethAb in animals receiving repeated Meth administration was still not known. In this study, we first investigated the tropism of AAV serotypes in rat primary dopaminergic (DA) neuronal culture. We found that AAV6 is an optimal gene carrier for MethAb. AAV6-MethAb or AAV6-mCherry was used in cellular and animal models of chronic Meth use. In primary DA neuronal culture, repeated Meth administration increased the dendritic branching of DA neurons, which was antagonized by AAV6-MethAb. AAV6-MethAb or AAV6-mCherry was stereotaxically administered to the nucleus accumbens (NAc) of adult CD1 mice. Two weeks after the viral injection, animals were stimulated with a daily dose of Meth for 7 days. Repeat Meth administrations led to a progressive increase in locomotor activity or behaviour sensitization. This response was significantly attenuated in mice receiving AAV6-MethAb. Using qRTPCR and Western analysis, we demonstrated that MethAb mRNA and protein were expressed in the NAc. Previous reports indicated that Meth sensitization was associated with upregulation of tyrosine hydroxylase (TH) in the NAc. Using Western blot analysis, we found that AAV6-MethAb significantly reduced TH protein levels in Meth-sensitized mice. Taken together, our data support that intracerebral administration of AAV6-MethAb reduced Meth sensitization. Our data support a novel antibody gene therapy for Meth abuse.
甲基苯丙胺（冰毒）是一种具有精神活性和神经毒性的化学物质。针对冰毒分子的选择性抗体已通过免疫接种用于治疗冰毒滥用。对冰毒具有高亲和力的抗体可以捕获冰毒分子并降低冰毒反应。我们之前报道过，腹腔注射携带冰毒特异性单克隆抗体转基因的8型腺相关病毒血清型载体（AAV8-MethAb，每只小鼠2.5×10病毒基因组拷贝数）可在外周诱导冰毒抗体的长期稳定表达。接受AAV8-MethAb的小鼠在急性注射冰毒后，血液和大脑中的冰毒水平较低，且冰毒诱导的运动活动减弱。AAV-MethAb对反复注射冰毒的动物的影响尚不清楚。在本研究中，我们首先研究了腺相关病毒血清型在大鼠原代多巴胺能（DA）神经元培养物中的嗜性。我们发现AAV6是MethAb的最佳基因载体。AAV6-MethAb或AAV6-mCherry被用于慢性使用冰毒的细胞和动物模型。在原代DA神经元培养中，反复注射冰毒增加了DA神经元的树突分支，而AAV6-MethAb可对抗这种作用。将AAV6-MethAb或AAV6-mCherry立体定向注射到成年CD1小鼠的伏隔核（NAc）中。病毒注射两周后，每天给动物注射一剂冰毒，持续7天。反复注射冰毒导致运动活动逐渐增加或行为敏感化。在接受AAV6-MethAb的小鼠中，这种反应明显减弱。通过qRTPCR和Western分析，我们证明了MethAb mRNA和蛋白在NAc中表达。先前的报道表明，冰毒敏感化与NAc中酪氨酸羟化酶（TH）的上调有关。通过Western印迹分析，我们发现AAV6-MethAb显著降低了冰毒敏感化小鼠中TH蛋白的水平。综上所述，我们的数据支持脑内注射AAV6-MethAb可降低冰毒敏感化。我们的数据支持一种针对冰毒滥用的新型抗体基因疗法。
