Development of cellular cancer immunotherapy by using humaniPS-cell-derived dendritic cells and ideal cancer-associated antigens

利用人PS细胞衍生的树突状细胞和理想的癌症相关抗原开发细胞癌症免疫疗法

基本信息

批准号：
23650609
负责人：
NISHIMURA Yasuharu
金额：
$ 2.33万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Challenging Exploratory Research
财政年份：
2011
资助国家：
日本
起止时间：
2011 至 2012
项目状态：
已结题

项目摘要

We established a method by which we can obtain a large number of functional dendritic cells (DC)with a simple procedure from human iPS cells. We transduced iPS cell-derived CD11b+myeloid cellswith cMYC and BMI1 genes associated with proliferative or anti-senescence effects. This made the cellscapable of propagating for more than 4 months in an M-CSF-dependent manner while retaining thecapacity to differentiate into DC. We named the iPS cell-derived proliferating myeloid cells “iPS-ML”,and the iPS-ML-derived dendritic cells “ML-DC”. In addition, we generated T AP2-deficient iPS cellclones by zinc finger nuclease-aided targeted gene disruption. T AP2-deficient iPS-ML avoidedrecognition by pre-activated allo-reactive CD8+T cells. The T AP2-deficient ML-DC expressingexogenously introduced HLA-A2 genes stimulated HLA-A2-restricted tumor antigen-specific CD8+Tcells obtained from HLA-A2-positive allogeneic donors, resulting in generation of tumorantigen-specific CTL lines. T AP-deficient iPS-ML introduced with various HLA class I genes may serveas an unlimited source of DC for vaccination therapy . Based on the present study , we propose aDC-producing system, which is simple, safe, and applicable to any patients irrespective of their HLAtypes. This technology will pave the way for the mass production of DC for therapeutic use. We havealso succeeded in inhibition of tumor growth of intraperitoneal dissemination of human cancer cells inimmune-deficient mice by intraperitoneal injection of human iPS-macrophage genetically modified tosecrete interferon-beta.

我们建立了一种方法，通过简单的程序可以从人 iPS 细胞中获得大量功能性树突状细胞 (DC)。我们用与增殖或抗衰老作用相关的 cMYC 和 BMI1 基因转导 iPS 细胞衍生的 CD11b+ 骨髓细胞。这使得细胞能够以 M-CSF 依赖性方式增殖超过 4 个月，同时保留分化为 DC 的能力，我们将其命名为 iPS 细胞源。增殖的骨髓细胞“iPS-ML”和iPS-ML衍生的树突状细胞“ML-DC”此外，我们通过锌指核酸酶辅助的靶向T AP2缺陷型iPS细胞克隆产生了T AP2缺陷型iPS细胞克隆。 -ML避免被预激活的同种异体反应性CD8+T细胞识别，表达外源引入的HLA-A2基因的T AP2缺陷ML-DC受到刺激。从 HLA-A2 阳性同种异体供体获得的 HLA-A2 限制性肿瘤抗原特异性 CD8+T 细胞，可产生引入各种 HLA I 类基因的肿瘤抗原特异性 CTL 系 T AP 缺陷 iPS-ML。基于目前的研究，我们提出了一种简单、安全且适用于任何患者的 DC 生产系统，无论其 HLA 类型如何。我们还通过腹腔注射经基因改造可分泌干扰素-β的人 iPS 巨噬细胞，成功地抑制了免疫缺陷小鼠腹腔内播散的人类癌细胞的肿瘤生长。