Gene Therapy with SLPI Promoter Controlled Replication-competent Adenovirus for Non-small Cell Lung Cancer

SLPI 启动子控制的具有复制能力的腺病毒对非小细胞肺癌的基因治疗

• 批准号: 15590793
• 负责人: MAEMONDO Makoto
• 金额: $ 2.24万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590793/
• 关键词: adenovirus; SLPI; NK4; gene therapy; angiogenesis; HGF; 複製可能アデノウイルス; 非小細胞肺癌

Secretory leukoprotease inhibitor (SLPI) is highly expressed in almost all non-small cell lung cancers (NSCLC), but not in the majority of other tumor types. In an attempt to create a specific gene therapy for NSCLC, we constructed AdSLPIE1AdB, an adenovirus vector with a double expression cassette consisting of E1A driven by the SLPI promoter gene followed by E1B-19K under the control of the CMV promoter that can selectively replicate only in NSCLC cells. Infection of AdSLPI.E1AdB yielded E1A protein expression and adenovirus replication resulting in a more than 100 fold increase of the virus titers only in SLPI-producing NSCLC cells (A549, H358, and HS24). In contrast, neither E1A protein nor replication was detected in non SLPI-producing HepG2 cells. Treatment with AdSLPI.EIAdB significantly inhibited the proliferation of NSCLC cells in vitro in a dose dependent manner, whereas the cell growth of HepG2 or normal human bronchial epithelial cells was not affected by AdSLPI.E1AdB infection. Direct injection of AdSLPI.E1AdB into A549 and H358 tumors in nude mice resulted in a marked reduction in tumor growth compared to controls (A549: 57%, p<0.02, H358: 67%, p<0.03). Histological examination revealed the replication of AdSLPI.E1AdB and strong induction of necrosis and apoptosis. In addition, we evaluated the combination of AdSLPI.E1AdB and AdCMVNK4 encoding NK4 protein which has strong anti-angiogenic activity E1A expressed by AdSLPI.E1AdB trans-acts on the replication of AdCMVNK4 and thus increases the expression of NK4. Injection of these two vectors into H358 tumors resulted i n a more striking reduction of tumor growth compare to single injection of each vector. These results suggest that AdSLPI.E1AdB could provide a selective therapeutic modality for NSCLC and that the combination of AdSLPI.E1AdB and AdCMVNK4 maybe a more effective gene therapy for NSCLC.

分泌性白细胞蛋白酶抑制剂 (SLPI) 在几乎所有非小细胞肺癌 (NSCLC) 中高表达，但在大多数其他肿瘤类型中不表达。为了创造针对 NSCLC 的特异性基因疗法，我们构建了 AdSLPIE1AdB，这是一种具有双表达盒的腺病毒载体，由 SLPI 启动子基因驱动的 E1A 和 CMV 启动子控制下的 E1B-19K 组成，只能在 NSCLC 细胞中选择性复制。 AdSLPI.E1AdB 的感染产生了 E1A 蛋白表达和腺病毒复制，导致仅在产生 SLPI 的 NSCLC 细胞（A549、H358 和 HS24）中病毒滴度增加了 100 倍以上。相比之下，在不产生 SLPI 的 HepG2 细胞中既没有检测到 E1A 蛋白，也没有检测到复制。 AdSLPI.EIAdB处理在体外以剂量依赖性方式显着抑制NSCLC细胞的增殖，而HepG2或正常人支气管上皮细胞的细胞生长不受AdSLPI.E1AdB感染的影响。与对照相比，将AdSLPI.E1AdB直接注射到裸鼠的A549和H358肿瘤中导致肿瘤生长显着减少（A549：57%，p<0.02，H358：67%，p<0.03）。组织学检查显示AdSLPI.E1AdB的复制以及对坏死和凋亡的强烈诱导。此外，我们评估了AdSLPI.E1AdB和编码NK4蛋白的AdCMVNK4的组合，AdSLPI.E1AdB表达的具有强抗血管生成活性的E1A反式作用于AdCMVNK4的复制，从而增加NK4的表达。与单次注射每种载体相比，将这两种载体注射到 H358 肿瘤中可更显着地减少肿瘤生长。这些结果表明AdSLPI.E1AdB可以为NSCLC提供选择性治疗方式，并且AdSLPI.E1AdB和AdCMVNK4的组合可能是NSCLC更有效的基因治疗。

项目成果

Gene therapy with secretory leukoprotease inhibitor promoter-controlled eplication-competent adenovirus for non-small cell lung cancer.

使用分泌性白蛋白酶抑制剂启动子控制的具有复制能力的腺病毒对非小细胞肺癌进行基因治疗。

• 发表时间: 2004
• 期刊: Cancer Res 64
• 作者: Ebihara T;Ebihara S;et al.;Maemondo M. et al.
• 通讯作者: Maemondo M. et al.

Pharmacokinetic analysis of combination chemotherapy with carboplatin and etoposide in small-cell lung cancer patients undergoing hemodialysis

• DOI: 10.1093/annonc/mdh008
• 发表时间: 2004-01-01
• 期刊: ANNALS OF ONCOLOGY
• 影响因子: 50.5
• 作者: Inoue, A;Saijo, Y;Nukiwa, T
• 通讯作者: Nukiwa, T

Adenovirus vector-mediated in vivo gene transfer of OX40 ligand to tumor cells enhances antitumor immunity of tumor-bearing hosts

• DOI: 10.1158/0008-5472.can-03-3911
• 发表时间: 2004-05-01
• 期刊: CANCER RESEARCH
• 影响因子: 11.2
• 作者: Andarini, S;Kikuchi, T;Nukiwa, T
• 通讯作者: Nukiwa, T

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Usui K、Saijo Y、Narumi k、Koyama S、Maemondo M 等人：“N 末端缺失增强了 BAX 在腺病毒基因递送至非小细胞肺癌中的细胞死亡诱导活性”Oncogene。

Suppression of peritoneal implantation of gastric cancer cells by adenovirus vector-mediated NK4 expression

• DOI: 10.1038/sj.cgt.7700782
• 发表时间: 2005-02
• 期刊: Cancer Gene Therapy
• 影响因子: 6.4
• 作者: H. Fujiwara;T. Kubota;H. Amaike;S. Inada;Kazuhiro Takashima;K. Atsuji;M. Yoshimura;M. Maemondo;K. Narumi;T. Nukiwa;Kunio Matsumoto;Toshikazu Nakamura;A. Hagiwara;H. Yamagishi