Gene therapy specific for lung cancer using cell-type specific promoter

使用细胞类型特异性启动子进行肺癌特异性基因治疗

• 批准号: 09670612
• 负责人: HAYASHI Seiji
• 金额: $ 1.6万
• 依托单位: OSAKA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670612/
• 关键词: Lung cancer; HSV-tk; CEA promoter; Cre; loxP system; myc oncogene; ganciclovir (GCV); nonproliferating adenoviral vector; 非増殖型アデノウイルスベクター

(1) Gene therapy specific for CEA-producing lung cancerRejection of CEA-producing tumors was not observed by intra-tumoral injection with a recombinant adenoviral vector (Ad.) expressing the HSV-tk gene under the CEA promoter (Ad.CEA-TK) followed by ganciclovir (GCV) treatment in vivo.It is conceivable that the low activity of the CPA promoter results in insufficient expression of the HSV-tk gene in cancer cells as well as in insignificant antitumor effects.To obtain enhanced expression of the HSV-tk gene exclusively in tumor cells and subsequent significant in vivo antitumor effects, we applied the Cre/loxP system to HSV-tk/GCV therapy for CEA-producing cancer.We constructed an Ad producing Cre recombinase driven by the CEA promoter (Ad.CEA-Cre) and another Ad designed for inducible expression of the HSV-tk gene by Cre (Ad.CAG-loxP-TK).Co-infection by these Ads rendered a human CEA-producing cancer cell line 8.4-fold more sensitive to GCV compared with infection by Ad.CEA-TK alone.On … More the other hand, co-infection with these Ads did not significantly change GCV sensitivity of CEA-non-producing cells.Intra-tumoral injection of Ad.CEA-Cre combined with Ad.CAG-loxP-TK followed by GCV treatment completely eradicated CEA-producing tumors established in the subcutis of 6 out of 7 athymic mice, whereas injection of Ad.CEA-TK alone with GCV administration at most retarded the growth of inoculated tumors.These results suggest distinct advantages of the Cre/loxP system applied in the conventional cell type-specific gene therapy against cancer.We are now evaluating the efficacy and side effects of this system in the peritonitis carcinomatosa model in mice, mimicking pleuritis for lung cancer in future clinical applications to human beings.(2) Gene therapy specific for Myc-overexpressing small cell lung cancer (Myc-SCLC)We previousely reported that an Ad. containing the HSV-tk gene ligated with the Myc-binding motif (Ad.Myc-TK) showed a cell-type specific expression of the HSV-tk gene in Myc-SCLC cell lines.We analyzed the efficacy of the intraperitoneal injection of Ad.Myc-TK followed by GCV treatment on Myc-SCLC inoculated in the peritoneal cavity of athymic mice.This treatment successfully reduced the weight of tumors to one fourteenth of that of untreated mice and completely eradicated tumors in 2 of 8 mice. Less

（1）未观察到针对CEA产生CEA产生CEA肿瘤的肺癌排斥的特异性基因治疗，并没有通过肿瘤内注射与重组腺病毒载体（AD。）表达HSV-TK基因在CEA启动子（AD.CEA-TK）下进行ganciclovir（gccv）治疗，以表达HSV-TK基因的vivo.IT conceiT conceiT con CONCE，启动子导致癌细胞中HSV-TK基因的表达不足以及在肿瘤细胞中仅在肿瘤细胞中获得HSV-TK基因的表达增强，随后在体内抗肿瘤效应中获得了显着性，我们将CRE/LOXP系统应用于HSV-TK/GCV Therapy。由CEA启动子（AD.CEA-CRE）和另一个用于通过CRE（AD.CAG-loxP-TK）诱导表达HSV-TK基因的AD。这些AD的co感染。与AD.CEA-TK相比，与GCV的感染相比，与其他人相比，与其他人相比，这些ADS使人类CEA癌细胞生成了8.4倍的癌细胞系更为敏感。 cea-non产生细胞的敏感性。对AD.CEA-CRE的肿瘤注射与AD.Cag-cag-loxP-TK结合，然后进行GCV处理，然后完全放射放射性的CEA产生肿瘤，在7个无壁画中，在7个无性小鼠中，在6个中，在6个小鼠中建立了aD.cea-cave tham tham tham thement in.cea-cave tham的范围，该肿瘤的增长量是在GCV的范围内。 advantages of the Cre/loxP system applied in the conventional cell type-specific gene therapy against cancer.We are now evaluating the effectiveness and side effects of this system in the peritonitis carcinomatosa model in mice, mimicking pleuritis for lung cancer in future clinical applications to human beings.(2) Gene therapy specific for Myc-overexpressing small cell lung cancer (Myc-SCLC)We previously reported that an广告。包含与MYC结合基序（AD.MYC-TK）结合的HSV-TK基因表明，在myc-SCLC细胞系中，HSV-TK基因的细胞类型的特异性表达表达。我们分析了AD.MYC-TK腹膜内注射的ATHIS治疗的perice clce cavity cavity cavity cavity cavity cavity cavity cavity在Perity cavity cavity cavity cavity cave insoneal cav insoneal cav insoneal cave insonean cave cave cave的效率进行了分析。在8只小鼠中有2只肿瘤的重量达到未处理小鼠的14个和完全放射的肿瘤。较少的

项目成果

Ueno K et al.: "Cloning and tissue expression of cDNAs from chromosome 5q21-22 which is frequently deleted in advanced lung cancer." Hum Genet. 102(1). 63-68 (1998)

Ueno K 等人：“染色体 5q21-22 cDNA 的克隆和组织表达，该染色体在晚期肺癌中经常被删除。”

Osaki T et al.: "Gene therapy for carcinoembryonic antigen-producing human lung cancer cells by cell type-specific expression of herpes simplex virus thymidine kinase gene." Cancer Res Oct. 15 ; 54(20). 5258-61 (1994)

Osaki T 等人：“通过单纯疱疹病毒胸苷激酶基因的细胞类型特异性表达，对产生癌胚抗原的人肺癌细胞进行基因治疗。”

Kumagai T et al.: "Eradication of Myc-overexpressing small cell lung cancer cells transfected with herpes simplex virus thymidine kinase gene containing Myc-Max response elements." Cancer Res Jan. 15 ; 56(2). 354-358 (1996)

Kumagai T 等人：“用含有 Myc-Max 反应元件的单纯疱疹病毒胸苷激酶基因转染的 Myc 过表达小细胞肺癌细胞的根除。”

Tachibana I et al.: "A 100-kDa protein tyrosine phosphorylation is concurrent with beta 1 integrin-mediated morphological differentiation in neuroblastoma and small cell lung cancer cells." Exp Cell Res Sep. 15 ; 227(2). 230-239 (1996)

Tachibana I 等人：“在神经母细胞瘤和小细胞肺癌细胞中，100 kDa 蛋白酪氨酸磷酸化与 β1 整合素介导的形态分化同时发生。”

Ueno K et al.: "Cloning and tissue expression of cDNAs from chromosome 5q21-22 which is frequently deleted in advanced lung cancer." Hum Genet Jan. 102(1). 63-8 (1998)

Ueno K 等人：“染色体 5q21-22 cDNA 的克隆和组织表达，该染色体在晚期肺癌中经常被删除。”