In vivo cancer gene therapy by adenovirus-mediated transfer of a bifunctional yeast cytosine deaminase/uracil phosphoribosyltransferase fusion gene.
In vivo cancer gene therapy by adenovirus-mediated transfer of a bifunctional yeast cytosine deaminase/uracil phosphoribosyltransferase fusion gene.
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发表时间：
2000-07
期刊：
影响因子：
11.2
通讯作者：
P. Erbs;E. Régulier;J. Kintz;P. Leroy;Y. Poitevin;F. Exinger;R. Jund;M. Mehtali
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作者：
P. Erbs;E. Régulier;J. Kintz;P. Leroy;Y. Poitevin;F. Exinger;R. Jund;M. Mehtali
将前药激活系统直接转移到肿瘤中是一种有吸引力的体内选择性清除肿瘤细胞的方法。然而，目前大多数自杀基因治疗策略仍然受到体内基因转移效率低和旁观者细胞杀伤效果有限的限制。在这项研究中，我们描述了一种新的高效自杀基因，该基因来源于酿酒酵母胞嘧啶脱氨酶(FCY1)和尿嘧啶磷酸核糖转移酶基因(FUR1)。这种命名为FCU1的自杀基因编码一种双功能嵌合蛋白，它结合了FCY1和FUR1的酶活性，并有效地催化无毒抗真菌药物5-FC直接转化为有毒代谢物5-氟尿嘧啶和5-氟尿嘧啶-5‘-单磷酸，从而绕过了某些人类肿瘤细胞对5-氟尿嘧啶的天然耐药性。出乎意料的是，尽管FCU1的尿嘧啶磷酸核糖转移酶活性与FUR1编码的相当，但其胞嘧啶脱氨酶活性却比FCY1编码的高出100倍。因此，转导表达FCU1的腺病毒(Ad-FCU1)的肿瘤细胞对5-FC的浓度比转导表达FCU1的载体(Ad-FCY1)的肿瘤细胞对5-FC的浓度低1000倍。此外，在转导Ad-FCU1的细胞中，旁观者细胞的杀伤作用也比单独或联合感染Ad-FCY1或Ad-FUR1的细胞更有效。最后，瘤内注射Ad-FCU1到同种或异种移植瘤中。在小鼠体内，伴随着全身注射5-FC，导致了肿瘤生长的显著延迟。这些独特的性质使FCU1/5-FC前体药物激活系统成为癌症基因治疗策略的新的和强大的候选者。
Direct transfer of prodrug activation systems into tumors was demonstrated to be an attractive method for the selective in vivo elimination of tumor cells. However, most current suicide gene therapy strategies are still handicapped by a poor efficiency of in vivo gene transfer and a limited bystander cell killing effect. In this study, we describe a novel and highly potent suicide gene derived from the Saccharomyces cerevisiae cytosine deaminase (FCY1) and uracil phosphoribosyltransferase genes (FUR1). This suicide gene, designated FCU1, encodes a bifunctional chimeric protein that combines the enzymatic activities of FCY1 and FUR1 and efficiently catalyzes the direct conversion of 5-FC, a nontoxic antifungal agent, into the toxic metabolites 5-fluorouracil and 5-fluorouridine-5'monophosphate, thus bypassing the natural resistance of certain human tumor cells to 5-fluorouracil. Unexpectedly, although the uracil phosphoribosyltransferase activity of FCU1 was equivalent to that encoded by FUR1, its cytosine deaminase activity was 100-fold higher than the one encoded by FCY1. As a consequence, tumor cells transduced with an adenovirus expressing FCU1 (Ad-FCU1) were sensitive to concentrations of 5-FC 1000-fold lower than the ones used for cells transduced with a vector expressing FCY1 (Ad-FCY1). Furthermore, bystander cell killing was also more effective in cells transduced with Ad-FCU1 than in cultures infected with Ad-FCY1 or Ad-FUR1, alone or in combination. Finally, intratumoral injections of Ad-FCU1 into allo- or xenogeneic tumors implanted s.c. into mice, with concomitant systemic administration of 5-FC, led to substantial delays in tumor growth. These unique properties make of the FCU1/5-FC prodrug activation system a novel and powerful candidate for cancer gene therapy strategies.