权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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，通过该方法，我们可以获得大量的功能性树突状细胞（DC）与一个简单的程序从人iPS细胞。我们用与增殖或抗衰老作用相关的cMYC和BMI 1基因转导iPS细胞来源的CD 11b+骨髓细胞。这使得细胞能够以M-CSF依赖的方式增殖超过4个月，同时保持向DC分化的能力。我们将iPS细胞衍生的增殖骨髓细胞命名为“iPS-ML”，将iPS-ML衍生的树突细胞命名为“ML-DC”。此外，我们通过锌指核酸酶辅助的靶向基因破坏来产生TAP 2缺陷的iPS细胞克隆。缺乏TAP 2的iPS-ML避免了预活化的同种异体反应性CD 8 +T细胞的识别。TAP 2缺陷型ML-DC表达外源性导入的HLA-A2基因，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CD 8 + T细胞，导致肿瘤抗原特异性CTL细胞系的产生。TAP-iPS-ML基因缺陷的iPS-ML可以作为一个无限的DC来源的疫苗接种治疗。基于目前的研究，我们提出了一个DC产生系统，这是简单的，安全的，适用于任何患者，无论他们的HLA型。这项技术将为大规模生产治疗用DC铺平道路。我们还成功地通过腹腔注射经基因修饰分泌干扰素β的人iPS-巨噬细胞，抑制了免疫缺陷小鼠中人癌细胞腹腔内扩散的肿瘤生长。