Evaluation of the therapeutic effect of replication-competent adenovirus on thyroid carcinomas

具有复制能力的腺病毒对甲状腺癌的治疗效果评价

• 批准号: 12671090
• 负责人: NAGAYAMA Yuji
• 金额: $ 2.18万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671090/
• 关键词: Thyroid carcinoma; Gene therapy; replication-competent adenovirus; 増殖型アデノウイルス; アデノウイルス; p53

We studied the therapeutic effect of the conditionally replication competent adenovirus on thyroid carcinomas in vitro and in vivo.(1) Differentiated thyroid carcinomas - We constructed the replicative adenovirus (AdTgE1A) in which the E1A gene, an indispensable gene for adenovirus replication, is placed downstream of thyroglobulin (Tg) promoter, a thyroid-specific promoter. Infection of AdTgElA led to E1A expression, virus replication (progeny production) and cytopathic effect in differentiated thyroid carcinoma cells expressing Tg, but not in other cells expressing no Tg. Furthermore, our studies demonstrated the enhanced effect of the combination of AdTgE1A and other gene therapies, including adenoviruses expressing a suicide gene or a cytokine.(2) Anaplastic thyroid carcinomas - To achieve p53-mutated cell-specific replication of adenovirus, we used "gene inactivation strategy" with p53-responseive promoter, Cre-LoxP system and double infection method. This strategy worked very well in vitro, but not so well in vivo, presumably because of low in vivo infectivity. Construction of a single adenovirus containing all the components was found impossible, since p53 promoter is not absolutely silent in 293 cells.

我们在体内外研究了条件复制型腺病毒对甲状腺癌的治疗作用：(1)分化型甲状腺癌-我们构建了复制型腺病毒(AdTgE1A)，其中E1a基因位于甲状腺特异性启动子甲状腺球蛋白(TG)启动子的下游，E1a基因是腺病毒复制必不可少的基因。AdTgElA感染诱导分化后表达TG的甲状腺癌细胞E1a表达、病毒复制(后代产生)和细胞病变效应，而不表达TG的其他细胞则不表达。此外，我们的研究证实了AdTgE1A与其他基因治疗的联合作用，包括表达自杀基因或细胞因子的腺病毒。(2)间变性甲状腺癌-为了实现腺病毒的p53突变细胞特异性复制，我们使用了带有p53反应启动子、Cre-loxP系统和双重感染的“基因失活策略”。这一策略在体外效果很好，但在体内效果不是很好，可能是因为体内传染性很低。由于P53启动子在293细胞中并不是绝对沉默的，因此构建包含所有成分的单一腺病毒是不可能的。

项目成果

Nishihara E., Nagayama Y., Inoue S., Hiroi H., Muramatsu M., Yamashita S., Koji T.: "Ontogenetic changes in the expression of estrogen receptor a and p in rat pituitary gland detected by immunohistochemistry"Endocrinology. 141(2). 615-620 (2000)

Nishihara E.、Nagayama Y.、Inoue S.、Hiroi H.、Muramatsu M.、Yamashita S.、Koji T.：“通过免疫组织化学检测大鼠垂体中雌激素受体 a 和 p 表达的个体发生变化”内分泌学。

Nagayama Y., Nishihara E., Namba H., Yamashita S., Niwa M.: "Identification of the sites of asparagine-linked glycosylation on human thyrotropin receptor and studies on their role in the receptor function and expression"J. Pharmacol. Exp. Ther.. 295 (1).

Nagayama Y.，Nishihara E.，Namba H.，Yamashita S.，Niwa M.：“人促甲状腺素受体上天冬酰胺连接糖基化位点的鉴定及其在受体功能和表达中的作用的研究”J。

Yuji Nagayama: "Targeting the replication of adenovirus to p53-defective thyroid carcinoma with a p53-regulated Cre-IoxP system."Cancer Gene Therapy. 8(1). 36-44 (2001)

Yuji Nagayama：“利用 p53 调节的 Cre-IoxP 系统将腺病毒复制靶向 p53 缺陷型甲状腺癌。”癌症基因治疗。

Eijum Nishihara: "Ontogenic changes in the expression of estrogen receptor α and β in rat pituitary gland detected by immunohistochemistry"Endocrinology. 141(2). 615-620 (2000)

Eijum Nishihara：“通过免疫组织化学检测大鼠垂体中雌激素受体α和β表达的个体变化”内分泌学141（2）（2000）。

Yuji Nagayama: "Adenovirus-mediated tumor suppressor p53 gene therapy for anaplastic throid carcinoma in vitro and in vivo"Journal of Clinical Endocrinology and Metabolism. 85(11). 4081-4086 (2000)

Yuji Nagayama：“腺病毒介导的肿瘤抑制p53基因治疗未分化甲状腺癌的体外和体内”临床内分泌与代谢杂志。