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In vivo HVJ-liposome mediated gene transfer into intestine and carcinoma.

体内 HVJ 脂质体介导的基因转移到肠和癌中。

基本信息

批准号：
08457336
负责人：
OKAMOTO Eizo
金额：
$ 5.12万
依托单位：
First Department of Surgery, Hyogo College 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：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457336/
关键词：
HVJ-liposome Intestine Cytokine Inflammatory bowel disease HGF HVJ リポソーム消化官 HVJリボゾーム

项目摘要

Several transfection methods have been developed to deliver exogenous genes into gastro-intestinal tract in vivo, but all have limitations. We evaluated the ability of Hemagglutmating virus of Japan (HVJ)-liposome mediated gene transfer to be used for gene therapy of incurrable intestinal diseases.We first investigated the possibilities of HVJ-liposome mediated gene trasfere in vivo. Fluorescent isothiocyanate (FTTC )-labeled oligodeoxynucleotides (ODN) and Escherichia Coli beta -galactosidase (beta-gal) gene was introduced into rat intestine by HVJ-liposome in vivo to examine transfer efficacy throughtwo approaches ; one was delivered via mesenteric artery and the other was intraluminal delivery after preparation with Pronase to remove mucus barrier, In the both method the localization of FITC-Labeled ODN and expression of beta-gal gene was observed not only in intestinal lamina propria but also muscle layer of the rat intestine. Our results demonstrate that HVJ-liposome method is useful for introduction of foreign gene into intestinal tract. Furthermore we established the model of hapten (TNB) induced chronic colitis as a target site of gene therapy, which mimic human inflammatory bowel disease (IBD). Prior to gene therapy, we analyzed various cytokines such as IL-12, IL-18, TNF-alpha, INF-gamma and determined role of cytokine in this model utilizing IL- 18 knock out mouse and neutralization antibody for IL- 18. We conclude that in this model TNF-alpha and INF-gamma seems to play a major role of inducing chronic colitis. To investigate possibilities of HVJ-liposome mediated gene therapy against inflammatory bowel disease we trasfer HGF in this model and it result in decreasing colitis. For the future study we are estabished a model of radiation colitis utilizing for gene thrapy.

目前已经开发了几种将外源基因转染到体内胃肠道的方法，但都有局限性。我们评估了日本血凝病毒（HVJ）脂质体介导的基因转移用于不可治愈的肠道疾病的基因治疗的能力。我们首先研究了hvj脂质体介导的基因在体内转移的可能性。采用hvj脂质体将荧光异硫氰酸酯（FTTC）标记的寡脱氧核苷酸（ODN）和大肠杆菌β -半乳糖苷酶（β -gal）基因导入大鼠肠道，通过两种途径检测其转移效果；一种是经肠系膜动脉给药，另一种是经Pronase去除黏液屏障后经腔内给药。两种方法不仅在大鼠肠固有层，而且在大鼠肠肌层都观察到了fitc标记的ODN的定位和β -gal基因的表达。结果表明，hvj -脂质体法是外源基因导入肠道的有效方法。此外，我们建立了半抗原（TNB）诱导的慢性结肠炎模型作为基因治疗的靶点，模拟人类炎症性肠病（IBD）。在基因治疗之前，我们分析了IL-12、IL-18、tnf - α、inf - γ等多种细胞因子，并利用IL-18敲除小鼠和IL-18中和抗体确定了细胞因子在该模型中的作用。我们得出结论，在这个模型中，tnf - α和inf - γ似乎在诱导慢性结肠炎中起主要作用。为了探讨hvj脂质体介导的基因治疗对炎症性肠病的可能性，我们在该模型中转移HGF，并导致结肠炎的减少。为了今后的研究，我们建立了一个利用基因治疗的放射性结肠炎模型。