Development of novel cancer cell vaccine using GM-CSF gene-transuduced IPS cells targeting cancer stem cells

使用GM-CSF基因转导的IPS细胞靶向癌症干细胞开发新型癌细胞疫苗

• 批准号: 23790446
• 负责人: INOUE Hiroyuki
• 金额: $ 2.66万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23790446/
• 关键词: 癌幹細胞; iPS細胞; 腫瘍免疫療法; GM-CSF; 遺伝子治療; 肺癌; 腫瘍免疫; ワクチン

Our results of in vitro assays demonstrated that non-transmissible recombinant Sendai virus-mediated mouse GM-CSF gene transfer to iPS (iPS/GM) cells was effective to produce abundant GM-CSF in vitro（200～ng/106cells/24 hrs）and iPS/GM-CSF cells maintained their stemness in terms of morphology and antigenicity as evidenced by the expression levels of SSEA-1,Oct3/4 and alkaline phosphatase similar to those seen in unmodified iPS cells. Our results of in vivo studies using immunocompetent mice demonstrated that mice treated with both prophylactic and therapeutic vaccination using irradiated iPS/GM-CSF (ir.iPS/GM-CSF) cells significantly suppressed the tumor growth of subcutaneously transplanted syngeneic LLC mouse poorly immunogenic lung cancer in the left flank. Of note, during these treatments described above, no serious adverse events were observed with lack of liver and kidney dysfunctions as evidenced by biochemical analysis as well as absence of loss of body weight, suggesting its relative tolerability of use of iPS cells as vaccine cells. Furthermore, in vivo depletion assays showed that the antitumor effect observed in mice treated with ir.iPS/GM-CSF cells was significantly abrogated by depleting CD4+T or CD8+T cells, demonstrating its partial dependence on T cell-mediated cellular antitumor immunity. In conclusion, this is the first report that demonstrated a therapeutic antitumor efficacy of normal fibroblast-derived iPS cell vaccines including iPS/GM cells, providing possible implications that these novel vaccine strategy using iPS cells could induce CSCs associated antigens-specific antitumor immunity and be a promising prophylactic or therapeutic anticancer modality

体外实验结果表明，非传染性重组仙台病毒介导的小鼠GM-CSF基因转移到iPS（iPS/GM）细胞中能有效地在体外产生大量的GM-CSF（200~ng/106个细胞/24小时）和iPS/GM-CSF细胞在形态学和抗原性方面保持了它们的干细胞性，如SSEA-1的表达水平所证明的，Oct 3/4和碱性磷酸酶与未修饰的iPS细胞中观察到的相似。我们使用免疫活性小鼠的体内研究结果表明，使用经辐照的iPS/GM-CSF（ir.iPS/GM-CSF）细胞进行预防性和治疗性疫苗接种处理的小鼠显著抑制了皮下移植的同基因LLC小鼠左胁腹中免疫原性差的肺癌的肿瘤生长。值得注意的是，在上述这些治疗期间，没有观察到严重的不良事件，如生化分析所证明的，没有肝和肾功能障碍，也没有体重减轻，这表明使用iPS细胞作为疫苗细胞的相对耐受性。此外，体内耗竭试验表明，在用ir.iPS/GM-CSF细胞处理的小鼠中观察到的抗肿瘤作用通过耗竭CD 4 +T或CD 8 +T细胞而显著消除，证明其部分依赖于T细胞介导的细胞抗肿瘤免疫。总之，这是第一份证明正常成纤维细胞衍生的iPS细胞疫苗（包括iPS/GM细胞）的治疗性抗肿瘤功效的报告，提供了使用iPS细胞的这些新型疫苗策略可以诱导CSC相关抗原特异性抗肿瘤免疫的可能含义，并且是有希望的预防或治疗性抗癌模式

项目成果

A novel cancer cell vaccine using induced pluripotent stem cells genetically engineered to produce GM-CSF elicits substantial antitumor immunity in a syngeneic mouse model

一种新型癌细胞疫苗，使用基因工程诱导多能干细胞产生 GM-CSF，在同基因小鼠模型中引发显着的抗肿瘤免疫力

• 发表时间: 2013
• 作者: Hiroyuki Inoue;Ayumi Watanabe,Chika Sakamoto;Megumi Narusawa;Shohei Miyamoto;Makoto Inoue;Koichi Takayama;Mamoru Hasegawa;Yo ichi Nakanishi;and Kenzaburo Tani.
• 通讯作者: and Kenzaburo Tani.

Genetically engineered oncolytic Edmonston strain of measles virus harboring the wild-type N, P, L Genes (MV-NPL) effectively target lung cancer stem cells.

基因工程麻疹病毒溶瘤埃德蒙斯顿株含有野生型 N、P、L 基因 (MV-NPL)，可有效靶向肺癌干细胞。

• 发表时间: 2012
• 作者: Ayumi Wanatabe;Hiroyuki Inoue;Chika Sakamoto;Megumi Narusawa;Shohei Miyamoto;Makoto Inoue;Keisuke Okita;Koichi Takayama;Mamoru Hasegawa;Yoichi Nakanishi;Shinya Yamanaka;and Kenzaburo Tani;Miho Nishizaki;井上博之
• 通讯作者: 井上博之

Coxsackievirus B3 Is an Oncolytic Virus with Immunostimulatory Properties That Is Active against Lung Adenocarcinoma

• DOI: 10.1158/0008-5472.can-11-3185
• 发表时间: 2012-05-15
• 期刊: CANCER RESEARCH
• 影响因子: 11.2
• 作者: Miyamoto, Shohei;Inoue, Hiroyuki;Tani, Kenzaburo
• 通讯作者: Tani, Kenzaburo

Novel cancer immunotherapy using induced pluripotent stem cells genetically engineered to produce GM-CSF

使用基因工程诱导多能干细胞产生 GM-CSF 的新型癌症免疫疗法

• 发表时间: 2012
• 作者: Ayumi Wanatabe;Hiroyuki Inoue;Chika Sakamoto;Megumi Narusawa;Shohei Miyamoto;Makoto Inoue;Keisuke Okita;Koichi Takayama;Mamoru Hasegawa;Yoichi Nakanishi;Shinya Yamanaka;and Kenzaburo Tani
• 通讯作者: and Kenzaburo Tani

Large-Scale Screening Identifies Coxsackievirus B3 (CVB3) as a Promising Oncolytic Virotherapy Agent against Non-Small Cell Lung Cancer

大规模筛选确定柯萨奇病毒 B3 (CVB3) 是一种有前景的针对非小细胞肺癌的溶瘤病毒治疗剂

• 发表时间: 2011
• 作者: Ayumi Wanatabe;Hiroyuki Inoue;Chika Sakamoto;Megumi Narusawa;Shohei Miyamoto;Makoto Inoue;Keisuke Okita;Koichi Takayama;Mamoru Hasegawa;Yoichi Nakanishi;Shinya Yamanaka;and Kenzaburo Tani;Miho Nishizaki;井上博之;井上博之
• 通讯作者: 井上博之