Do retroviral vector insertions influence the fate of terminally differentiated cells such as T lymphocytes by insertional mutagenesis?

逆转录病毒载体插入是否会通过插入突变影响 T 淋巴细胞等终末分化细胞的命运？

• 批准号: 22728825
• 负责人: Professor Dr. Boris Fehse
• 金额: --
• 依托单位: Interdisziplinäre Klinik für Stammzelltransplantation
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/22728825?language=en
• 关键词: Do; retroviral; vector; insertions; influence

Gene transfer with (y-)retroviral (RV) vectors into hematopoietic stem and progenitor cells may heavily influence the fate of single cells eventually leading to clonal dominance or even malignant transformation. The observed survival advantage of those clones has been attributed to random vector insertions into proto-oncogenes or other signaling genes. However, it is not known yet whether such strong side effects of retroviral vector insertions may also be expected with terminally differentiated cells. We here aim at investigating this question using peripheral T cells, an important target of various gene therapy strategies. We will perform dose-escalated T cell transduction with both y-retro and lentiviral vectors and analyze the long-term impact of insertion sites in vitro and in vivo, in murine BMT models. Vectors with strong promotor/enhancer elements developed by our partner Baum will be used to mimic a worst case scenario . In cooperation with other applicants of this SPP1230 (Frühauf, von Laer, von Kalle) we will also analyze RV vector insertions in T cells from clinical gene therapy samples. Together this data should allow to get novel insights into the risk of malignant transformation of T lymphocytes by both y-retro- and lentiviral gene transfer. Through a network of cooperation this project will also significantly contribute to several of the common platforms of the given SPP, incl. insertional genomics, models of clonat selection, mathematical modeling and vectorology.

使用（γ）逆转录病毒（RV）载体将基因转移到造血干细胞和祖细胞中可能会严重影响单细胞的命运，最终导致克隆优势甚至恶性转化。观察到的这些克隆的生存优势归因于随机载体插入原癌基因或其他信号基因。然而，尚不清楚逆转录病毒载体插入的如此强烈的副作用是否也适用于终末分化细胞。我们的目的是利用外周 T 细胞来研究这个问题，外周 T 细胞是各种基因治疗策略的重要目标。我们将使用 y-retro 和慢病毒载体进行剂量递增的 T 细胞转导，并在小鼠 BMT 模型中分析体外和体内插入位点的长期影响。我们的合作伙伴 Baum 开发的具有强启动子/增强子元件的载体将用于模拟最坏的情况。我们还将与该 SPP1230 的其他申请人（Frühauf、von Laer、von Kalle）合作，分析来自临床基因治疗样本的 T 细胞中的 RV 载体插入情况。综合这些数据，我们可以对 y 逆转录病毒和慢病毒基因转移引起的 T 淋巴细胞恶性转化的风险有新的认识。通过合作网络，该项目还将为特定 SPP 的多个通用平台做出重大贡献，包括。插入基因组学、克隆选择模型、数学建模和载体学。