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Basic Research of Gene Therapy for Joint Diseases

关节疾病基因治疗基础研究

基本信息

批准号：
09470320
负责人：
KUBO Toshikazu
金额：
$ 5.76万
依托单位：
KYOTO PREFECTURAL UNIVERSITY OF MEDICINE
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-09470320/
关键词：
gene therapy adenovirus vector guinea pig in vivo drug delivery system TGF-betaI joint 変形性関節症アデノウイルスペクター LacZ 軟骨細胞プロモーター

项目摘要

We evaluated the in vivo applicability of adenovirus-mediated gene delivery in order to consider the feasibility of gene therapy for human joint diseases.We directly injected vectors harbouring beta-galactosidase (beta-gal) or transforming growth factor (TGF)-beta1 gene into the joints of Hartley guinea pigs. Expressions of transduced beta-gal genes were examined by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining and reverse transcription- polymerase chain reaction (RT-PCR). The levels of TGF-beta1 which was delivered to the joint and then transferred to the joint fluid, were assessed by enzyme-linked immunosorbent assay (ELISA). LacZ expression was observed in almost the entire synovial tissues and in chondrocytes on the surface of degenerated cartilage. Un expected effects of the direct vector inj ection on the other organs were also examined, no expression of delivered gene was observed. Therefore, intraarticular direct injection would achieve gene delivery limited to the joint cavity, possibly eliminating unecessary systemic effects. TGF-beta1 levels in the joint fluid following the gene delivery had been significantly higher than the levels in the controls for 2 weeks.Direct gene delivery into the joint cavity is realistic with the in vivo gene delivery method using adenovirus vector, and it would be clinically applicable.

我们评估了腺病毒介导的基因传递在体内的适用性，以考虑基因治疗人类关节疾病的可行性。我们将含有β -半乳糖苷酶（β -gal）或转化生长因子(TGF)- β 1基因的载体直接注射到哈特利豚鼠关节内。通过5-溴-4-氯-3-吲哚- β - d -半乳糖苷（X-gal）染色和逆转录-聚合酶链反应（RT-PCR）检测转导的β -gal基因的表达。通过酶联免疫吸附试验（ELISA）评估tgf - β 1的水平，tgf - β 1被递送到关节，然后转移到关节液中。LacZ几乎在整个滑膜组织和退变软骨表面的软骨细胞中表达。我们还检测了直接载体注射对其他器官的意外影响，未观察到传递基因的表达。因此，关节内直接注射将实现基因在关节腔内的传递，可能消除不必要的全身效应。基因传递后2周关节液中的tgf - β 1水平显著高于对照组。利用腺病毒载体的体内基因传递方法直接将基因送入关节腔是可行的，具有临床应用价值。