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

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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.