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Pain treatment with xenogeneic cells transfected with drug inducible opiate gene

用药物诱导阿片基因转染的异种细胞治疗疼痛

基本信息

批准号：
09557118
负责人：
SAITOH Youichi
金额：
$ 3.71万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557118/
关键词：
pain beta-endorphin polymer capsule ex vivo gene therapy xenograft Tet-On system tetracycline doxycycline Pain オピエ-トテトラサイクリンポリマーカプセル異種細胞

项目摘要

We previously reported that xenogeneic cells, carrying the proopiomelanocortin (POMC) gene coding the precursor of the ACTH and 13-endorphin in a polymer capsule transplanted into the cerebrospinal fluid (CSF) space of rats, secrete b-endorphin and reduce their pain. However, the activities of transplanted cells were barely controlled by exogenous agents. The Tet-On system, one of the transcriptional transactivators, which requires certain tetracycline derivatives for specific DNA binding, has been developed and considered to be applicable for the control of the transplanted cells. The pUHD1O-3-POMC and pUHD172-lneo were co-transfected to Neuro2A, a mouse neuroblastoma cell line, named Neuro2A-POMC.Neuro2A cells carrying the doxycycline-inducible POMC gene expressed and secreted ACTH upon addition of the doxycycline in vitro in a dose-dependent manner. The encapsulated and transplanted cells in the CSF space of rats were stimulated by four intraperitoneal injections of doxycycline, and found to express ACTH after subsequence 36-hour incubation in vitro, depending on the amounts of injected doxycycline. These findings indicate that gene expression in xenogeneic cells in the CSF space can be manipulated by injection a relatively harmless drug, and suggest that our system is applicable to cell transplantation therapy in central nervous system diseases requiring well-controlled substance delivery.

我们以前报道，异种细胞，携带前阿黑皮素（POMC）基因编码的前体的促肾上腺皮质激素和13-内啡肽的聚合物胶囊移植到大鼠的脑脊液（CSF）空间，分泌β-内啡肽，并减少他们的疼痛。然而，移植细胞的活性几乎不受外源性试剂的控制。Tet-On系统是转录反式激活因子之一，它需要某些四环素衍生物进行特异性DNA结合，已被开发并被认为适用于移植细胞的控制。将pUHD 10 -3-POMC和pUHD 172-lneo共转染到小鼠神经母细胞瘤细胞系Neuro 2A中，命名为Neuro 2A-POMC，在体外加入强力霉素后，携带强力霉素诱导型POMC基因的Neuro 2A细胞以剂量依赖的方式表达和分泌ACTH。在大鼠CSF空间的封装和移植的细胞被刺激的四个多西环素的腹腔注射，并发现表达促肾上腺皮质激素后，在体外培养36小时，这取决于注射的多西环素的量。这些研究结果表明，异种细胞在CSF空间中的基因表达可以通过注射相对无害的药物来操纵，并表明我们的系统适用于需要良好控制的物质递送的中枢神经系统疾病的细胞移植治疗。