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

基本信息

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

项目摘要

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.

分泌的白细胞抑制剂（SLPI）在几乎所有非小细胞肺癌（NSCLC）中高度表达，但在大多数其他肿瘤类型中均未表达。为了创建针对NSCLC的特定基因疗法，我们构建了ADSLPIE1ADB，这是一种腺病毒矢量，具有双表达盒，由SLPI启动子基因驱动的E1A组成，然后由E1B-19K，然后是CMV启动子的控制，可以选择性地重复出现。在NSCLC细胞中。 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，病毒滴度仅增加了100倍的病毒滴度。相反，在非SLPI产生的HEPG2细胞中均未检测到E1A蛋白和复制。用ADSLPI.EIADB治疗以剂量依赖性的方式显着抑制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。与对照组相比，裸鼠中直接注射ADSLPI.E1ADB中裸鼠的A549和H358肿瘤导致肿瘤生长显着降低（a549：57％，p <0.02，h358：67％，p <0.03）。组织学检查表明，ADSLPI.E1ADB的复制以及坏死和凋亡的强诱导。此外，我们评估了编码NK4蛋白的ADSLPI.E1ADB和ADCMVNK4的组合，该蛋白具有强烈的抗血管生成活性E1A，ADSLPI.E1ADB trans-acts在ADCMVNK4的复制中表达了E1A，从而增加了NK4的表达。将这两个向量注射到H358肿瘤中导致肿瘤生长的降低与单次注射相比，肿瘤生长的降低更加明显。这些结果表明，ADSLPI.E1ADB可以为NSCLC提供选择性的治疗方式，并且ADSLPI.E1ADB和ADCMVNK4的组合可能是NSCLC的更有效的基因治疗。