Basic study for. Cancer Gene Therapy by Antitumor Effects Using Fas-Fas Ligand-Mediated Apoptosis

基础学习。

• 批准号: 12671144
• 负责人: TAKEDA Yasutaka
• 金额: $ 2.11万
• 依托单位: The University of Tokyo
• 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-12671144/
• 关键词: FasL(CD95L)-transfected tumor cell; Cancer gene therapy; Apoptosis; Anti-tumor effect; Neutrophil infiltration; MH134 hepatoma; Anti-Fas antibody; Resistant tumor; Fasリガンド発現腫瘍; 抗腫瘍作用; 肝癌MH134; FasL発現腫瘍; MH134

In the immunologically privileged sites, FasL (ligand) is constitutively expressed and causes prompt apoptosis of infiltrating lymphoid cells. FasL can protect the grafted tissue from immune-mediated damage. However, we and other researchers reported that various types of tumors, when genetically manipulated to express FasL, were rejected after inducing marked inflammation with extensive neutrophil infiltrate. Thus, we investigated cancer therapy using FasL-transfected tumor cells.Four kinds of FasL cDNA-transfected murine tumor cells (neuroblastoma Neuro-2a, lung carcinoma 3LL, hepatoma MH134, fibrosarcoma MethA) were rejected when injected into mice, and induced strong antitumor immunity. Thus, the antitumor activity of FasL did not depend on tumor type. To address how FasL-expressing tumors induce neutrophil emigration and abrogate tumorigenicity, we investigated the behavior of G2 (MHl34+FasL) cells injected into +/+, lpr^<cg>/lpr^<cg>(lpr^<cg> and gld/gld Ipr/lpr (gld/lpr) mice. G … More 2 cells were eradicated after extensive infiltration of neutrophils around them in +/+ mice but formed tumors without such infiltration in lpr^<cg> and gld/lpr mice. These results indicate that apoptosis of neutrophils with FasL-expressing tumors depends on Fas and may trigger the extensive infiltration of neutrophils resulting in violent inflammation and ultimately in eradication of tumor cells in + mice. The antitumor effect was examined by mixing G2 cells with MH134 cells (model of Fas/FasL-negative tumor) or F6b (MH134+Fas) and injecting into mice. The mixture of tumor cells was eradicated after extensive infiltration of neutrophils around them. The antitumor effect of G2 cells against F6b cells was extremely stronger than that against parent cells (MH134). These results suggested that FasL-expressing tumor exerted antitumor activity against un-transfected tumor cells by-stander effects. This seems to be useful for the clinical application, because all tumor cells could not be always transfected with transfected cDNA. On the other hand, the suppression of resistant cells which appear in treatment is important for augmentation in tumor-gene therapy. To solve this problem, we examined the antitumor effect of F6b cells and anti-Fas Mab-resisant F6b cells by anti-Fas Mab. The resistant cells could survive by modulating Fas cDNA which resulted in the decrease or the depletion of Fas expression for the attack by the anti-Fas Mab.Collectively, our results indicate that transfection of FasL into tumor cells is a good method for the induction of strong antitumor effects and might be much useful for clinical application. Less

在免疫豁免部位，FasL（配体）组成型表达，并导致浸润淋巴细胞的迅速凋亡。FasL可以保护移植组织免受免疫介导的损伤。然而，我们和其他研究人员报道，当基因操作表达FasL时，各种类型的肿瘤在诱导显著炎症和广泛的中性粒细胞浸润后被排斥。因此，我们研究了使用FasL转染的肿瘤细胞的癌症治疗，四种FasL cDNA转染的小鼠肿瘤细胞（神经母细胞瘤Neuro-2a，肺癌3LL，肝癌MH 134，纤维肉瘤MethA）注射到小鼠体内时被排斥，并诱导了强的抗肿瘤免疫。因此，FasL的抗肿瘤活性不依赖于肿瘤类型。为了解决表达FasL的肿瘤如何诱导嗜中性粒细胞移出并消除致瘤性，我们研究了注射到+/+、lpr^<cg>/lpr^<cg>（lpr^）<cg>和gld/gld Ipr/lpr（gld/lpr）小鼠中的G2（MH 134 +FasL）细胞的行为。G ...更多信息 在+/+小鼠中，2细胞在其周围的嗜中性粒细胞广泛浸润后被根除，但在lpr+和gld/lpr小鼠中形成肿瘤而没有这种浸润<cg>。这些结果表明，FasL表达肿瘤的中性粒细胞凋亡依赖于Fas，并可能引发中性粒细胞的广泛浸润，导致剧烈炎症，并最终根除+小鼠中的肿瘤细胞。通过将G2细胞与MH 134细胞（Fas/FasL阴性肿瘤模型）或F6 b（MH 134 +Fas）混合并注射到小鼠中来检查抗肿瘤效果。肿瘤细胞周围广泛浸润中性粒细胞后，肿瘤细胞的混合物被根除。G2细胞对F6 b细胞的抗肿瘤作用明显强于亲本细胞（MH 134）。这些结果表明，表达FasL的肿瘤对未转染的肿瘤细胞具有旁观者效应的抗肿瘤活性。这似乎是有用的临床应用，因为所有的肿瘤细胞不能总是转染的cDNA。另一方面，抑制治疗中出现的耐药细胞对于增强肿瘤基因治疗是重要的。为了解决这一问题，我们研究了F6 b细胞和抗Fas单克隆抗体的F6 b细胞的抗肿瘤作用。耐药细胞可通过调节Fas cDNA的表达，使Fas表达降低或缺失，从而抵抗抗Fas单抗的攻击，从而存活下来。以上结果表明，FasL转染肿瘤细胞是一种诱导强抗肿瘤效应的良好方法，具有重要的临床应用价值。少

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