Recognition of Glioma Stem Cells by Genetically Modified T Cells Targeting EGFRvIII and Development of Adoptive Cell Therapy for Glioma
Recognition of Glioma Stem Cells by Genetically Modified T Cells Targeting EGFRvIII and Development of Adoptive Cell Therapy for Glioma
复制标题
DOI：
10.1089/hum.2012.041
复制
发表时间：
2012-10-01
影响因子：
4.2
通讯作者：
Rosenberg, Steven A.
中科院分区：
文献类型：
--
作者：
Morgan, Richard A.;Johnson, Laura A.;Rosenberg, Steven A.
胶质母细胞瘤目前尚无根治疗法，确诊后中位生存期不到2年。作为开发胶质母细胞瘤免疫治疗方法的一种方法，我们试图以胶质瘤干细胞(GSCs)中表达的抗原为靶点。与已建立的胶质瘤细胞系相比，GSC具有多种特性，使其在胶质瘤肿瘤中更具代表性。表皮生长因子受体变异体III(EGFRvIII)是一种新的肿瘤特异性基因重排的结果，它产生了一种在大约30%的胶质瘤中表达的独特蛋白，是免疫治疗的理想靶点。使用跨越EGFRvIII特异性缺失的聚合酶链式反应，我们发现这个肿瘤特异性基因在三个GCS系中的三个中表达。根据7个EGFRvIII特异性单抗的序列信息，我们组装了嵌合抗原受体(CAR)，并评价了CAR工程T细胞识别EGFRvIII的能力。这些抗EGFRvIII基因工程T细胞中有三个产生了效应细胞因子干扰素-c和裂解抗原表达的靶细胞。我们集中开发了由人mAb139产生的CAR，它特异性地识别表达突变的EGFRvIII的GSC系和胶质瘤细胞系，但不识别野生型EGFR，并且不识别任何被测试的正常人类细胞。使用基于139的CAR，来自胶质母细胞瘤患者的T细胞可以通过基因工程来识别表达EGFRvIII的肿瘤，并可以在体外大量扩增，并保持其抗肿瘤活性。在此基础上，构建了表达EGFRvIII CAR的c-逆转录病毒载体，用于临床应用。
No curative treatment exists for glioblastoma, with median survival times of less than 2 years from diagnosis. As an approach to develop immune-based therapies for glioblastoma, we sought to target antigens expressed in glioma stem cells (GSCs). GSCs have multiple properties that make them significantly more representative of glioma tumors than established glioma cell lines. Epidermal growth factor receptor variant III (EGFRvIII) is the result of a novel tumor-specific gene rearrangement that produces a unique protein expressed in approximately 30% of gliomas, and is an ideal target for immunotherapy. Using PCR primers spanning the EGFRvIII-specific deletion, we found that this tumor-specific gene is expressed in three of three GCS lines. Based on the sequence information of seven EGFRvIII-specific monoclonal antibodies (mAbs), we assembled chimeric antigen receptors (CARs) and evaluated the ability of CAR-engineered T cells to recognize EGFRvIII. Three of these anti-EGFRvIII CAR-engineered T cells produced the effector cytokine, interferon-c, and lysed antigen-expressing target cells. We concentrated development on a CAR produced from human mAb 139, which specifically recognized GSC lines and glioma cell lines expressing mutant EGFRvIII, but not wild-type EGFR and did not recognize any normal human cell tested. Using the 139-based CAR, T cells from glioblastoma patients could be genetically engineered to recognize EGFRvIII-expressing tumors and could be expanded ex vivo to large numbers, and maintained their antitumor activity. Based on these observations, a c-retroviral vector expressing this EGFRvIII CAR was produced for clinical application.