Gene therapy against gynecologic cancers targeting telomerase

针对端粒酶的妇科癌症基因治疗

• 批准号: 09470354
• 负责人: INOUE Masaki
• 金额: $ 8.45万
• 依托单位: Kanazawa University
• 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 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470354/
• 关键词: Gynecologic tumors; Telomere; Telomerase; hTERT; hTERT-promoter; Gene therapy; 産婦人科腫瘍; TRAP法; 転写因子; hTRT

Human telomerase is composed of three main subunit components, human telomerase RNA (hTR), telomerase-associated protein (TP1) and telomerase reverse transcriptase (hTERT). RT-PCR analyses demonstrated that expression of hTERT was observed in cancer tissues and cell lines, while hTR or TP1 was broadly expressed not only in cancers but also in normal tissues. Thus, hTERT expression was well correlated with telomerase activity.We succeeded to clone promoter sequences of hTERT gene. Of particular importance is identification of transcription factors interacting with such regions and regulate hTERT express in utilizing it for gene therapy. C-Myc has been shown to be a direct transactivator of hTERT gene. Antisense strategy against c-Myc has been in success to inhibit telomerase activity in cancer cells and suppress the tumor growth. As another strategy, we have applied hTERT promoter to plasmid or virus vectors for gene therapy. We have constructed a chimeric vector in which hTERT promoter is cloned upstream of apoptosis-inducing genes inhibiting cell growth and introduced of these vectors into cervical cancer cells. We have confirmed the specific inhibition of tumor growth in cell lines and animal models by these gene therapies.

人端粒酶由三个主要亚基成分：人端粒酶RNA（HTR），端粒酶相关蛋白（TP1）和端粒酶逆转录酶（HTERT）组成。 RT-PCR分析表明，在癌症组织和细胞系中观察到HTERT的表达，而HTR或TP1不仅在癌症中，而且在正常组织中也广泛表达。因此，HTERT表达与端粒酶活性很好地相关。我们成功地克隆了HTERT基因的启动子序列。特别重要的是鉴定与此类区域相互作用的转录因子并调节HTERT Express将其用于基因治疗。 C-MYC已被证明是HTERT基因的直接反式激活剂。针对C-MYC的反义策略已成功地抑制癌细胞中的端粒酶活性并抑制肿瘤生长。作为另一种策略，我们已将HTERT启动子应用于基因治疗的质粒或病毒载体。我们已经构建了一个嵌合载体，其中将HTERT启动子克隆到凋亡诱导基因的上游抑制细胞生长，并将这些载体引入宫颈癌细胞中。我们已经通过这些基因疗法证实了细胞系和动物模型中肿瘤生长的特异性抑制作用。

项目成果

Kyo S,et al: "hTERT as a critical determinant of telomerase activity in normal and malignant endometrium" Int J Cancer. 80. 60-63 (1998)

Kyo S 等人：“hTERT 作为正常和恶性子宫内膜中端粒酶活性的关键决定因素”Int J Cancer。

Kyo S et al.: "Telomerase activity in cervical cancer is quantitatively distinct from that in its precursors" In J Cancer. 79. 66-70 (1998)

Kyo S 等人：“宫颈癌中的端粒酶活性在数量上与其前体癌中的端粒酶活性不同”，发表在 J Cancer 中。

Kyo, S. et al.: "Estrogen activates telomerase"Cancer Res. 59. 5917-5921 (1999)

Kyo, S. 等人：“雌激素激活端粒酶”癌症研究。

Kyo S, Takakura M, Kanaya T, Taira T, Ithoh H, Yutsudo M, Ariga H, Inoue M.: "Sp1 cooperates with c-myc to activate transcription of human telomerase reverse transcriptase (hTERT) gene."Nucleic Acid Res.. (In press.). (2000)

Kyo S、Takakura M、Kanaya T、Taira T、Ithoh H、Yutsudo M、Ariga H、Inoue M.：“Sp1 与 c-myc 配合激活人端粒酶逆转录酶 (hTERT) 基因的转录。”核酸研究。

Takakura, M., Kyo, S. et al.: "Cloning of human telomerase catalytic subunit gene promoter"Cancer Res. 59. 551-557 (1999)

Takakura, M., Kyo, S.等：“人端粒酶催化亚基基因启动子的克隆”Cancer Res。