In vivo PET measurement of expression of dopamine D2 receptor gene injected into rat striatum

体内PET测量大鼠纹状体注射多巴胺D2受体基因的表达

• 批准号: 10558115
• 负责人: ISHIWATA Kiichi
• 金额: $ 8.45万
• 依托单位: TOKYO METROPOLITAN INSTITUTE OF GERONTOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10558115/
• 关键词: dopamine D2 receptor; gene transfer; positron emission tomography; adenovirus; raclopride; autoradiography; MRI; C-11; アデノウィルス; ドパミン受容体

As the basic study of gene therapy, we investigated whether the expression of dopamine DィイD22ィエD2 receptor (DィイD22ィエD2R) gene injected into the rat striatum was evaluated in vivo by positron emission tomography (PET). preliminarily we established that the striatal DィイD22ィエD2R was evaluated by PET with ィイD111ィエD1C-labeled DィイD22ィエD2R-specific ligands by using normal rats and a rat model for striatal neurodegenerative diseases induced by striatal injection of quinolinic acid. To overcome the spatial resolution of PET camera, a technique of PET-MRI registration on the rat brain was developed.PET measurement demonstrated that the uptake of three kinds of DィイD22ィエD2R-specific ligands, [ィイD111ィエD1C]raclopride, [ィイD111ィエD1C]nemonapride and [ィイD111ィエD1C]N-methylspiperone was higher in the striatum injected with the vectors for DィイD22ィエD2R than the contralateral striatum injected with a control vector 2-3 days after injection. The uptake of [ィイD111ィエD1C]SCH 23390, a dopamine DィイD21ィエD2 receptor … More specific ligand, or [ィイD111ィエD1C]β-CIT-FP, a dopamine transporter specific tracer, was not different between bilateral striata, and co-injection of excess unlabeled raclopride inhibited the uptake of [ィイD111ィエD1C]raclopride, suggesting the PET signal reflects the DィイD22ィエD2R-specific gene-expression. These findings were confirmed by ex vivo and in vitro autoradiography. At day 16 the increased uptake of [ィイD111ィエD1C]raclopride declined to basal level, suggesting that the gene expression is temporary.We concluded that PET imaging is capable of detecting adenovirus-mediated gene transfer in vivo. This technique represents a major advance over in vitro ARG analysis of DィイD22ィエD2R binding which must be done post-hoc. PET imaging permits in vivo analysis of the efficiency of DィイD22ィエD2R gene transfer that can be followed longitudinally and related to any functional changes being observed. Such imaging will prove highly valuable for assessment of gene transfer using this and similar vectors. Less

作为基因治疗的基础研究，我们用正电子发射断层扫描技术观察了多巴胺D-ィイ-D22-ィエ-D2受体(D-ィイ-D22-ィエ-D2R)基因注入大鼠纹状体后的体内表达情况。我们初步建立了纹状体DィイD22ィエD2R与ィイD111ィエD1C标记的DィイD22ィエD2R特异性配体的正电子发射计算机断层扫描。为了克服正电子发射计算机成像的空间分辨率问题，建立了一种大鼠脑内正电子发射计算机断层成像配准技术。PET测量表明，注射DィイD22ィエD2R载体2~3天后，[ィイD111ィエD1C]、[ィイD111ィエD1C]Nemonaprib和[ィイD111ィエD1C]N-甲基螺丙酮3种DィイD22ィエD2R特异性配体在纹状体的摄取率高于注射对照载体的对侧。[ィイD111ィエD1C]SCH 23390，一种多巴胺DィイD21ィエD2受体…的摄取多巴胺转运体特异性示踪剂[ィイD111ィエD1C]β-Cit-FP在双侧纹状体间无差异，联合注射过量的未标记雷氯普利可抑制[ィイD111ィエD1C]的摄取，提示正电子发射计算机断层扫描信号反映了DィイD22ィエD2R特异性基因的表达。这些发现得到了体外和体外放射自显影的证实。在第16天，[ィイD111ィエD1C]雷克洛必利摄取的增加下降到基础水平，表明基因表达是暂时的。这项技术代表了对DィイD22ィエD2R结合的体外ARG分析的重大进步，后者必须在HOC后完成。PET成像可以在体内分析DィイD22ィエD2R基因转移的效率，这种基因转移可以进行纵向跟踪，并与观察到的任何功能变化有关。这种成像将被证明对使用该载体和类似载体进行基因转移的评估具有很高的价值。较少

项目成果

Ishiwata K., Ogi N., Tanaka A. and Senda M.: "Quantitative ex vivo and in vitro receptor autoradiography using ィイD111ィエD1C-labeled ligands and an imaging plate: a study with a dopamine DィイD22ィエD2-like receptor ligand [ィイD111ィエD1C]nemonapride"Nucl. Med. Bi

Ishiwata K.、Ogi N.、Tanaka A. 和 Senda M.：“使用 D111D1C 标记的配体和成像板进行定量离体和体外受体放射自显影：使用多巴胺 D22D D2 样受体配体 [D111D1C]nemonapride 的研究“核医学双

Hayakawa N., Uemura K., Ishiwata K., Shimada Y., Ogi N., Nagaoka T., Toyama H., Oda K., Tanaka A., Endo K. and Senda M.: "A PET-MRI registration technique for PET studies of the rat brain"Nucl. Med. Biol.. (in press).

Hayakawa N.、Uemura K.、Ishiwata K.、Shimada Y.、Ogi N.、Nagaoka T.、Toyama H.、Oda K.、Tanaka A.、Endo K. 和 Senda M.：“PET-MRI 配准

K Ishiwata et al.: "Quantitative ex vivo and in vitro autoradiographyusing ^<11>C-labeled lignds and imaging plate : a study with a dopamine D2-like receptor ligand [^<11>C]nemonapride"Nuclear Medicine and Biology. 26. 291-296 (1999)

K Ishiwata 等人：“使用 ^ 11 C 标记配体和成像板进行定量离体和体外放射自显影：使用多巴胺 D2 样受体配体 [^ 11 C]nemonapride 进行的研究”核医学和生物学。

K. Ishiwata et al.: "Comparison of three PET dopamine D_2-like receptor ligands, [^<11>C] raclopride, [^<11>C] nemonapride and [^<11>C] N-methylspiperone, in rats"Annals of Nuclear Medicine. 13. 161-167 (1999)

K. Ishiwata 等人：“三种 PET 多巴胺 D_2 样受体配体 [^11C] 雷氯必利、[^11C] 奈莫必利和 [^11C] N-甲基螺哌隆在大鼠体内的比较

N. Hayakawa et al.: "A PET-MRI registration technique for PET studies of the rat brain"Nuclear Medicine and Biology. (in press).

N. Hayakawa 等人：“用于大鼠大脑 PET 研究的 PET-MRI 配准技术”核医学与生物学。