Gene Delivery and Addiction

重组腺相关病毒（rAAV）载体经常用于基因传递到中枢神经系统，并且能够转导神经元。利用表达胶质源性神经营养因子（GDNF）的AAV载体，我们能够与我们的校内合作者Yavin Shaham博士一起为一项研究做出贡献。我们发现，在可卡因戒断后的第一天注射含有大鼠GDNF cDNA的腺相关病毒（AAV）病毒载体会增加在戒断后第11天和第31天线索诱导的可卡因寻找；使用对照病毒时没有观察到这种效果。完整的研究考察了GDNF在可卡因渴求的孵化过程中的作用。详细的研究目前正在审查中，准备发表。

Recombinant adeno-associated viral (rAAV) vectors are frequently used for gene delivery to the central nervous system and are capable of transducing neurons. Using an AAV vector expressing glial derived neurotrophic factor (GDNF), we were able to contribute to a study with our intramural collaborator, Dr. Yavin Shaham. We show that injections of an adeno-associated virus (AAV) viral vector containing rat GDNF cDNA on the first day after cocaine withdrawal increased cue-induced cocaine-seeking on withdrawal days 11 and 31; this effect was not observed using a control virus. The complete study examines the role of GDNF in the incubation of cocaine craving. The detialed study is currently under review for publication. One of the proteins that our section has been studying for many years is bone morphogenetic protein 7, specifically, its neurotrophic/neuroregenerative properties. We previously demonstrated that exogenous application of bone morphogenetic protein 7 (BMP7) reduced 6-hydroxydopamine-mediated neurodegeneration in a rodent model of Parkinson's disease. We published a study this past year showing that transgenic mice expressing a truncated form of the BMP receptor II in dopaminergic neurons of the nigrostriatal pathway were more vulnerable to methamphetamine challenge as determined by TUNEL labeling in the substantia nigra. Our data suggest that a deficiency in BMP signaling increases vulnerability to insults induced by high doses of MA. In a second study, we show that methamphetamine suppressed BMP7 expression in the substantia nigra of mice. Mice that carry only one allele of BMP7 were more vulnerable to methamphetamine toxicity as measured by tyrosine hydroxylase (TH) immunostaining in the nigra reticulate and changes in locomotor activity. Exogenous delivery of BMP7 protein via the lateral ventricle of mouse brain reduced methamphetamine toxicity as measured by locomotor activity and TH immunostaining. Collectively, our data indicate that BMP7 and BMP receptor signaling has neuroprotective effects against methamphetamine toxicity in the nigrostriatal system. We recently generated an AAV vector expressing the human mu opioid receptor (huMOR) and have begun evaluating the role of huMOR in methamphetamine sensitization in mice. Our preliminary findings show that huMOR expression by an AAV vector in specific brain regions alters methamphetamine sensitization. This work is ongoing. An AAV vector expressing the glutamate transporter (GLT-1) was created to modulated the levels of extracellular glutamate. We have begun experiments examining the ability of excess GLT-1 overexpression by AAV to reduce excitoxicity by glutamate. We have also begun examining the GLT-1 overexpression in specfic brain regions for alterations to methamphetamine sensitization. Once fully characterized, the AAV-GLT1 vector will be used in other models of drug sensitization, craving and reward.